2024/11/01 更新

写真a

イマムラ コレヨシ
今村 維克
IMAMURA Koreyoshi
所属
環境生命自然科学学域 教授
職名
教授
外部リンク

学位

  • 工学博士 ( 1999年6月   京都大学 )

  • 工学修士 ( 京都大学 )

研究キーワード

  • 材料物理化学

  • 生物化学工学

  • 乾燥工学

  • Food Technology

  • Chemical Engineering

  • 食品工学

  • 化学工学

研究分野

  • ものづくり技術(機械・電気電子・化学工学) / バイオ機能応用、バイオプロセス工学  / タンパク質,疎水性薬剤,酵素反応

  • ライフサイエンス / 薬系分析、物理化学  / アモルファス固体分散,ガラス転移

  • ライフサイエンス / 食品科学  / 食品工学

  • ものづくり技術(機械・電気電子・化学工学) / 移動現象、単位操作  / 乾燥,非晶質糖類,固体分散

学歴

  • 京都大学    

    - 1997年

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    国名: 日本国

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  • 京都大学   Graduate School, Division of Engineering  

    - 1997年

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  • 京都大学   Faculty of Engineering  

    - 1992年

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  • 京都大学   Faculty of Engineering  

    - 1992年

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    国名: 日本国

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経歴

  • 岡山大学   学術研究院環境生命自然科学学域創成化学講座応用化学コース   教授   博士(工学)

    2023年4月 - 現在

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  • 岡山大学   学術研究院自然科学学域応用化学専攻   教授   博士(工学)

    2021年4月 - 2023年3月

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    国名:日本国

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  • 岡山大学   大学院自然科学研究科   教授

    2012年6月 - 2021年3月

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  • 岡山大学   大学院自然科学研究科   准教授

    2005年4月 - 2012年5月

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  • 岡山大学   工学部   助教授

    2003年12月 - 2005年3月

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  • - Cleaning of solid surfaces using radical

    1998年

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  • 岡山大学   工学部   助手

    1997年4月 - 2003年11月

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  • - Adsorption of protein, peptide, &

    1997年

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  • - Stabilization of protein by sugar

    1995年

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  • oxidizing reaction

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  • amino acidly auto solid surfaces

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▼全件表示

所属学協会

委員歴

  • 化学工学会男女共同参画委員会   副委員長  

    2023年4月 - 現在   

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    団体区分:学協会

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  • 日本食品工学会   理事  

    2017年7月 - 現在   

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    団体区分:学協会

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  • 日本生物工学会   西日本支部会計  

    1999年   

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    団体区分:学協会

    日本生物工学会

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  • 日本食品工学会   理事  

    1906年1月   

 

論文

  • 噴霧乾燥および保存時におけるアモルファス固体分散中の疎水性薬剤の結晶化阻害に関する研究 査読

    Yamamoto, Rina, Imanaka, H, Ishida, Naoyuki, Imamura, Koreyoshi

    Drying Technology   42 ( 13 )   2022 - 2031   2024年11月

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    担当区分:最終著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    添付ファイル: P0SF-Cry-draft.pdf

    DOI: 10.1080/07373937.2024.2401502

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  • 種々の条件における真空foam乾燥および噴霧乾燥によって調製したアモルファス固体分散体の香料保持特性 査読

    Nitta, Yuna, Sato, H, Yamamoto, Rina, Imanaka, Hiroyuki, Ishida, Naoyuki, Imamura, K

    42 ( 2 )   227 - 237   2024年1月

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    担当区分:最終著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    添付ファイル: P0SF-Flavor-draft.pdf

    DOI: 10.1080/07373937.2023.2285413

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  • Water sorption and glass-to-rubber transition of amorphous sugar matrices, vacuum foam- and spray-dried from alcohols 査読

    Koji Takeda, Shinta Miyazaki, Takashi Okamoto, Hiroyuki Imanaka, Naoyuki Ishida, Koreyoshi Imamura

    JOURNAL OF FOOD ENGINEERING   349   2023年7月

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    担当区分:最終著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCI LTD  

    The objective of this study is to understand the impact of preparation conditions on the physicochemical characteristics of amorphous sugar. Amorphous sugar (alpha-maltose, isomaltulose, trehalose) matrices were pre -pared from alcohol (methanol, ethanol) solutions, and their water sorption behavior and glass transition tem-peratures, Tg, were compared with those of aqueous freeze-dried amorphous sugar. Vacuum-foam-and spray -drying procedures were employed as drying methods. Water sorption for amorphous sugars, dried by different methods, was generally significant in the following order: vacuum-foam-drying < spray-drying <= freeze-drying. In the same order, the Tg values for the thoroughly dried amorphous sugars increased, and water sorption reduced the differences in Tg for given amounts of sorbed water. The solvent type in vacuum-foam-drying slightly affected the water sorption and the dependence of Tg on the sorbed water amount. The obtained findings sug-gested that physicochemical characteristics of amorphous sugar may relate to the sugar molecule conformations in solvent and dried state.

    添付ファイル: JFOODENG-D-22-02113_R3-3.pdf

    DOI: 10.1016/j.jfoodeng.2023.111483

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  • Evaluation of interaction forces between hydrophobic surfaces in surfactant solutions by atomic force microscopy 査読

    Shota Kage, Koreyoshi Imamura, Naoyuki Ishida

    Colloid and Polymer Science   301 ( 7 )   775 - 782   2023年7月

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    掲載種別:研究論文(学術雑誌)  

    The interaction forces between silica surfaces with various degrees of hydrophobicity were measured using colloidal probe atomic force microscopy to clarify the relationship between the hydrophobicity of the surfaces and hydrophobic attraction in detail. The interaction forces between silica surfaces pre-hydrophobised with octadecyltrichlorosilane were measured in aqueous solutions with three different (cationic, anionic, and nonionic) surfactants. Hydrophobic attraction was observed between surfaces from 10 to 15 nm without surfactant, and the range of this attraction decreased when the surface hydrophobicity decreased with increasing surfactant concentration. The hydrophobic attraction disappeared when the surface contact angle was below 80°–90° in the presence of all surfactants used in this study. The range of the hydrophobic attraction was found to depend strongly on the contact angle and not on the surfactant type and concentration. The adhesion force was also highly dependent on the contact angle and comparable to the theoretical capillary force. The possible origin of this force is discussed with reference to the obtained results. Graphical Abstract: [Figure not available: see fulltext.].

    DOI: 10.1007/s00396-023-05118-w

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  • Evaluation of Hydrophobic Attraction between Polystyrene Layer and Silanated Silica Surface by Atomic Force Microscopy 査読

    Yuta Okuda, Koreyoshi Imamura, Naoyuki Ishida

    Chemistry Letters   52 ( 12 )   878 - 882   2023年

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    掲載種別:研究論文(学術雑誌)  

    In this study, hydrophobic surfaces were prepared by depositing polystyrene (PS) on a glass substrate using three different methods to measure the interaction forces against a hydrophobized particle by atomic force microscopy. We found that smooth surfaces prepared by spin coating and living radical polymerization generated hydrophobic attraction, whereas uneven surfaces obtained by free radical polymerization produced only steric repulsion, even though they had sufficient hydrophobicity. Based on these results, the origin of the hydrophobic attraction was suggested.

    DOI: 10.1246/cl.230363

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  • Kinetics of Enzymatic Reactions at the Solid/Liquid Interface in Nanofluidic Channels 査読

    Koki Yamamoto, Kyojiro Morikawa, Hiroyuki Imanaka, Koreyoshi Imamura, Takehiko Kitamori

    Analytical Chemistry   2022年10月

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    掲載種別:研究論文(学術雑誌)   出版者・発行元:American Chemical Society (ACS)  

    DOI: 10.1021/acs.analchem.2c02878

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  • Extraordinary high preservation of the dispersion state of Au nanoparticles during freeze-thawing and freeze-drying with gum arabic 査読

    Miki Kadowaki, Tsutashi Matsuura, Hiroyuki Imanaka, Naoyuki Ishida, Koreyoshi Imamura

    Colloids and Surfaces A: Physicochemical and Engineering Aspects   639   2022年4月

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    担当区分:最終著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    A series of edible and non-toxic substances were screened for their ability to prevent Au nanoparticles (AuNPs) from undergoing aggregation during both freeze-thawing and freeze-drying at as low an additive concentration as possible. Gum arabic was found to be an ideal choice. The addition of 20 µg/mL of gum arabic greatly inhibited the aggregation of AuNPs during freeze-thawing and -drying, whereas polyvinylpyrrolidone was the next best among the tested substances, with approximately 20% aggregation during freeze-drying at a concentration of less than 200 µg/mL. The zeta potential value of the AuNPs indicated the absence of the specific binding of gum arabic to the AuNPs surface. Surface enhanced infrared spectroscopic (SEIRAS) analyses were conducted for the gum arabic solution, frozen and freeze-dried on the Au particles. The results indicated that, in the frozen state, gum arabic molecules were concentrated in remaining unfrozen liquid on the Au coating without any specific orientation toward the Au surface. In contrast, SEIRAS spectra of freeze-dried gum arabic indicated that carboxylic groups in gum arabic molecules preferentially became closer to the Au surface. Considering these findings, during freeze-thawing, we conclude that the high thickening property of gum arabic gum plays an important role in minimizing the mobility and thus the aggregation of AuNPs in the freeze-concentrated regions between frozen ice crystals. The freeze-drying of a gum arabic solution produces a rigid amorphous matrix that has an affinity for the Au surface. The AuNPs were individually separated and immobilized in the freeze-dried matrix of the gum arabic, resulting in the extraordinary inhibition of their aggregation.

    添付ファイル: COLSUA_128392_edit_report.pdf

    DOI: 10.1016/j.colsurfa.2022.128392

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  • Comparison of improvements of aqueous dissolution of structurally analogous hydrophobic drugs by amorphous solid dispersion 招待 査読

    Takashi Okamoto, Kayoko Yamamoto, Takanari Sekitoh, Akiho Fujioka, Hiroyuki Imanaka, Naoyuki Ishida, Koreyoshi Imamura

    Colloids and Surfaces A: Physicochemical and Engineering Aspects   632   127744 - 127744   2022年1月

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    担当区分:最終著者, 責任著者   掲載種別:研究論文(学術雑誌)   出版者・発行元:Elsevier BV  

    添付ファイル: P0SF-IBPs-draft.pdf

    DOI: 10.1016/j.colsurfa.2021.127744

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  • Induction of foaming in vacuum drying by needle stimulation and the impact of solution viscosity, vapor pressure, and the type of solute and solvent 査読

    Akiho Fujioka, Rina Yamamoto, Olivier Tramis, Hiroaki Ishida, Tsutomu Ono, Naoyuki Ishida, Koreyoshi Imamura

    Drying Technology   1 - 13   2021年12月

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    担当区分:最終著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Informa UK Limited  

    添付ファイル: P0NeedleEx-draft.pdf

    DOI: 10.1080/07373937.2021.2017965

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  • Foaming characteristics of sugar- and polyvinylpyrrolidone-alcohol solutions during vacuum foam drying: A rheological approach 査読

    Olivier Tramis, Akiho Fujioka, Hiroyuki Imanaka, Naoyuki Ishida, Koreyoshi Imamura

    Colloids and Surfaces A: Physicochemical and Engineering Aspects   627   127174 - 127174   2021年10月

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    担当区分:最終著者   記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Elsevier BV  

    Vacuum foam drying offers great perspectives to formulate solid-state encapsulated active drugs. Taking into account the specific need of pharmaceutical formulations to keep drug molecules active and dispersed, we show in this paper that vacuum drying of pharmaceutical formulations can be substantially improved by using original rheological approaches. Typically, our formulations totally dried-up in less than 5 min, a delay much shorter than the long hours of the traditional vacuum drying process. Steady-shear rheology was used to evaluate the solution's specific viscosity against the solutions' concentrations, in order to correlate the dilution regime to the foamabiltiy. This rheological approach indicated that in miscible solutions, spontaneous foaming occurred in the semi-dilute entangled regime, near the transition to the concentrated regime; for partially miscible solutions, it occurred in solutions at a concentration near to the percolation concentration. The proposed methodology is versatile, and should provide a simple way to assess the foamability of pharmaceutical formulations.

    添付ファイル: P0NeedleTramis2-pub.pdf

    DOI: 10.1016/j.colsurfa.2021.127174

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  • Inhibiting Au nanoparticle aggregation in freeze-thawing by presence of various additives 査読

    Miki Kadowaki, Hidetaka Yokota, Hiroyuki Imanaka, Naoyuki Ishida, Koreyoshi Imamura

    Advanced Powder Technology   32 ( 10 )   3517 - 3524   2021年10月

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    担当区分:最終著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Elsevier BV  

    添付ファイル: P1ParticleEx-draft.pdf

    DOI: 10.1016/j.apt.2021.08.002

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  • Direct measurements of interaction forces of bovine serum albumin and lysozyme with stainless steel by atomic force microscopy 査読

    Hikaru Kawaguchi, Hiroyuki Imanaka, Koreyoshi Imamura, Naoyuki Ishida

    Colloids and Surfaces A: Physicochemical and Engineering Aspects   627   127137 - 127137   2021年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Elsevier {BV}  

    DOI: 10.1016/j.colsurfa.2021.127137

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  • Sole-amorphous-sugar-based solid dispersion of curcumin and the influence of formulation composition and heat treatment on the dissolution of curcumin 査読

    Takanari Sekitoh, Takashi Okamoto, Akiho Fujioka, Olivier Tramis, Koji Takeda, Tsutashi Matsuura, Hiroyuki Imanaka, Naoyuki Ishida, Koreyoshi Imamura

    Drying Technology   39 ( 14 )   2065 - 2074   2021年9月

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    担当区分:最終著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Informa UK Limited  

    An amorphous solid dispersion technique, in which hydrophobic drug molecules are dispersed in an amorphous matrix comprised solely of a sugar, was applied to curcumin that has excellent physiological functions. The sole-amorphous-sugar-based solid dispersion (SAS-SD) of curcumin was prepared by vacuum foam drying from methanol, using various disaccharides as the drug carrier-forming component. Aqueous dissolution of curcumin from the SAS-SD showed “spring-and-parachute” shape profile and was markedly increased when a sugar (α-maltose or trehalose) was added in a composition range ≥100 g-sugar/g-curcumin. Further dissolution was achieved by heating the SAS-SD at close to and above the melting points of curcumin.

    添付ファイル: P0SF-Cur-draft.pdf

    DOI: 10.1080/07373937.2020.1752711

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  • アルコールから乾燥調製したトレハロースアモルファスの結晶化特性 査読

    関藤孝成, 岡本崇, 藤岡亜希穂, 吉岡朋彦, 照井真司, 今中洋行, 石田尚之, 今村維克

    292   110325 - 110325   2021年3月

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    担当区分:最終著者, 責任著者   掲載種別:研究論文(学術雑誌)  

    添付ファイル: P0TreCry-pub.pdf

    DOI: 10.1016/j.jfoodeng.2020.110325

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  • Direct measurement of interaction force between hydrophilic silica surfaces in triblock copolymer solutions with salt by atomic force microscopy 査読

    Takumi Deguchi, Takato Nakahara, Koreyoshi Imamura, Naoyuki Ishida

    Advanced Powder Technology   32 ( 1 )   30 - 36   2021年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Elsevier BV  

    DOI: 10.1016/j.apt.2020.11.011

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  • ポリビニルピロリドンおよび糖のアルコール溶液の真空乾燥における自発的な発泡と難水溶性薬物の対水溶解性の改善 査読

    Olivier Tramis,藤岡亜希穂,今中洋行,石田尚之,今村維克

    1 - 11   2020年9月

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    担当区分:最終著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    添付ファイル: P0NeedleTramisPVP.pdf

    DOI: 10.1080/07373937.2020.1822863

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  • 食品タンパク質をはじめとする生体分子の 包括安定化技術に関する工学研究 招待 査読

    今村 維克

    日本食品工学会誌   21 ( 3 )   95 - 111   2020年9月

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    担当区分:筆頭著者, 最終著者, 責任著者   記述言語:日本語   掲載種別:研究論文(学術雑誌)  

    添付ファイル: 200815食工研究賞原稿.pdf

    DOI: 10.11301/jsfe.20579

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  • A suitable and effective stepwise oxidative refolding procedure for highly‐cationic tetrameric avidin in nucleic acid free conditions 査読 国際誌

    Shuichiro Kimura, Koreyoshi Imamura, Junichiro Futami

    Biotechnology Progress   36 ( 5 )   e3031   2020年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Wiley  

    Optimized conditions are needed to refold recombinant proteins from bacterial inclusion bodies into their biologically active conformations. In this study, we found two crucial requirements for efficient refolding of cationic tetrameric chicken avidin. The first step is to eliminate nucleic acid contaminants from the bacterial inclusion body. The electrostatic interactions between the remaining nucleic acids and proteins strongly enhanced protein aggregation during the refolding process. The cysteine specific reversible S-cationization procedure was successfully employed for large-scale preparation of nucleic acid free denatured protein without purification tag system. The second step is the intramolecular disulfide formation prior to refolding in dialysis removing denaturant. Disulfide intact monomeric avidin showed efficient formation of biologically active tetrameric conformation during the refolding process. Using this optimized refolding procedure, highly cationic avidin derivative designed as an intracellular delivery carrier of biotinylated protein was successfully prepared.

    DOI: 10.1002/btpr.3031

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    その他リンク: https://onlinelibrary.wiley.com/doi/full-xml/10.1002/btpr.3031

  • 糖および界面活性剤複合系による金ナノ粒子の凍結乾燥における分散安定化 査読

    横田秀隆, 門脇美季, 松浦傳史, 今中洋行, 石田尚之, 今村維克

    36 ( 24 )   6698 - 6705   2020年6月

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    担当区分:最終著者, 責任著者   掲載種別:研究論文(学術雑誌)  

    添付ファイル: P1Particle-pub.pdf

    DOI: 10.1021/acs.langmuir.0c00695

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  • 外部電場による表面非親和性タンパク質の金属表面への固定化 査読

    129 ( 3 )   348 - 353   2020年3月

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    担当区分:最終著者, 責任著者   掲載種別:研究論文(学術雑誌)  

    添付ファイル: P2Tramis-pub.pdf

    DOI: 10.1016/j.jbiosc.2019.09.008

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  • バルクでの反応速度を超えるナノ流体デバイス上のピコリットル酵素反応装置 査読

    The Analyst   145 ( 17 )   5801 - 5807   2020年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1039/d0an00998a

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  • Pico-liter protein digestion and separation using nanofluidic device

    Kyojiro Morikawa, Koki Yamamoto, Hiroki Sano, Yutaka Kaoze, Hisashi Shimizu, Hiroyuki Imanaka, Koreyoshi Imamura, Takehiko Kitamori

    MicroTAS 2020 - 24th International Conference on Miniaturized Systems for Chemistry and Life Sciences   1125 - 1126   2020年

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    記述言語:英語   掲載種別:研究論文(国際会議プロシーディングス)   出版者・発行元:Chemical and Biological Microsystems Society  

    The Needs of shotgun proteomics have been increasing in the field of biology and medicine for understanding biological systems. In study, protein sample with pL, which volume was same scale as single cell volume, was successfully digested and separated and detected by integrated device of pL enzyme reactor fL chromatography and UV-POPS detector. This nanofluidic device will greatly contribute to single cell shotgun proteomics.

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  • ニードル刺激によるfoaming誘導を利用した低真空下における真空foam乾燥の乾燥履歴の制御 査読

    日高史博, 佐藤朋, 藤岡亜希穂, 竹田昂司, 今中洋行, 石田尚之, 今村維克

    37 ( 12 )   1520 - 1527   2019年9月

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    担当区分:最終著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    添付ファイル: LDRT_A_1517363draft.pdf

    DOI: 10.1080/07373937.2018.1517363

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  • アモルファス固体分散体としてメタノールから乾燥した糖類アモルファスマトリクスの物理的安定性と熱処理の影響 査読

    竹田昂司, 関藤孝成, 藤岡亜希穂, 山本佳代子, 岡本崇, 松浦傳史, 今中洋行, 石田尚之, 今村維克

    Journal of Pharmaceutical Sciences   108 ( 6 )   2056 - 2062   2019年6月

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    担当区分:最終著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    添付ファイル: P0TgOrganic-pub.pdf

    DOI: 10.1016/j.xphs.2019.01.008

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  • Nanofluidic enzyme reactor exceeding limit of bulk reaction rate

    Koki Yamamoto, Kyojiro Morikawa, Koreyoshi Imamura, Hiroyuki Imanaka, Takehiko Kitamori

    23rd International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2019   113 - 114   2019年

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    記述言語:英語   掲載種別:研究論文(国際会議プロシーディングス)   出版者・発行元:Chemical and Biological Microsystems Society  

    We development a nanofluidic enzyme reactor exceeding the limitation of bulk enzyme reaction. The limit concentration of bulk trypsin solution is approximately 0.5 PM because self-digestion occurs over 0.5 PM solution, which complicates analysis of target proteins by digestion trypsin itself. Using extremely high surface/volume ration of nanospace, trypsin concentration, which indicates number of immobilized trypsin molecules on the nanochannel surface for liquid phase volume, exceed to bulk limit of concentration. As a result, 31 times higher concentration and 33 times faster enzyme reaction compared to bulk was realized. This reactor will be powerful tools which overcomes limitation in bulk and microfluidic method.

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  • Adsorption characteristics of various proteins on a metal surface in the presence of an external electric potential 査読

    Colloids and Surfaces B: Biointerfaces   166   262 - 268   2018年6月

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    担当区分:最終著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    添付ファイル: pagination_COLSUB_9236.pdf

    DOI: 10.1016/j.colsurfb.2018.03.035

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  • Hydrophobic Attraction Measured between Asymmetric Hydrophobic Surfaces 査読

    Naoyuki Ishida, Kohei Matsuo, Koreyoshi Imamura, Vincent S. J. Craig

    Langmuir   34 ( 12 )   3588 - 3596   2018年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:American Chemical Society  

    The interaction forces between silica surfaces modified to different degrees of hydrophobicity were measured using colloidal probe atomic force microscopy (AFM). A highly hydrophobic silica particle was prepared with octadecyltrichlorosilane (OTS), and the interaction forces were measured against silica substrates modified to produce surfaces of varying hydrophobicity. The interaction forces between the highly hydrophobic particle and a completely hydrophilic silicon wafer surface fitted well to the DLVO theory, indicating that no additional (non-DLVO) forces act between the surfaces. When the silicon wafer surface was treated to produce a contact angle of water on surface of 40°, an additional attractive force that is longer ranged than the van der Waals force was observed between the surfaces. The range and magnitude of the attractive force increase with the contact angle of water on the substrate. Beyond the effect on the contact angle, the hydrocarbon chain length and the terminal groups of hydrophobic layer on the substrate only have a minor effect on the magnitude of the force, even when the substrate is terminated with polar carboxyl groups, provided the hydrophobicity of the other surface is high.

    DOI: 10.1021/acs.langmuir.7b04246

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  • Influence of an external electric field on removal of protein fouling on a stainless steel surface by proteolytic enzymes 査読

    COLLOIDS AND SURFACES B-BIOINTERFACES   159   118 - 124   2017年11月

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    担当区分:最終著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

    添付ファイル: P3Enzyme-pub.pdf

    DOI: 10.1016/j.colsurfb.2017.07.074

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  • Characteristics of proteinaceous additives in stabilizing enzymes during freeze-thawing and -drying 査読

    Takanori Shimizu, Tamayo Korehisa, Hiroyuki Imanaka, Naoyuki Ishida, Koreyoshi Imamura

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY   81 ( 4 )   687 - 697   2017年4月

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    担当区分:最終著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:TAYLOR & FRANCIS LTD  

    Protein-stabilizing characteristics of sixteen proteins during freeze-thawing and freeze-drying were investigated. Five enzymes, each with different instabilities against freezing and dehydration, were employed as the protein to be stabilized. Proteinaceous additives generally resulted in greater enzyme stabilization during freeze-thawing than sugars while the degree of stabilization for basic lysozyme and protamine were inferior to that of neutral and acidic proteins. Freeze-drying-induced inactivation of enzyme was also reduced by the presence of a proteinaceous additive, the extent of which was lower than that for a sugar. In both freeze thawing and freeze drying, the enzymes stabilization by the proteinaceous additive increased with increasing additive concentration. The enhancement of enzyme inactivation caused by pH change was also reduced in the presence of proteinaceous additives. The combined use of a sugar such as sucrose and dextran tended to increase the stabilizing effect of the proteinaceous additive.

    添付ファイル: TBBB_A_1274637r1.pdf

    DOI: 10.1080/09168451.2016.1274637

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  • Surfactant-Free Solid Dispersions of Hydrophobic Drugs in an Amorphous Sugar Matrix Dried from an Organic Solvent 査読

    Koji Takeda, Yuto Gotoda, Daichi Hirota, Fumihiro Hidaka, Tomo Sato, Tsutashi Matsuura, Hiroyuki Imanaka, Naoyuki Ishida, Koreyoshi Imamura

    MOLECULAR PHARMACEUTICS   14 ( 3 )   791 - 798   2017年3月

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    担当区分:最終著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER CHEMICAL SOC  

    The technique for homogeneously dispersing hydrophobic drugs in a water-soluble solid matrix (solid dispersion) is a subject that has been extensively investigated in the pharmaceutical industry. Herein, a novel technique for dispersing a solid, without the need to use a surfactant, is reported. A freeze-dried amorphous sugar sample was dissolved in an organic solvent, which contained a soluble model hydrophobic component. The suspension of the sugar and the model hydrophobic component was vacuum foam dried to give a solid powder. Four types of sugars and methanol were used as representative sugars and the organic medium. Four model drugs (indomethacin, ibuprofen, gliclazide, and nifedipine) were employed. Differential scanning calorimetry analyses indicated that the sugar and model drug (100:1) did not undergo segregation during the drying process. The dissolution of the hydrophobic drugs in water from the solid dispersion was then evaluated, and the results indicated that the C-max and AUC(0-60) (min) of the hydrophobic drug in water were increased when the surfactant-free solid dispersion was used. Palatinose and/or alpha-maltose were superior to the other tested carbohydrates in increasing C-max and AUC(0-60 min), for all tested model drugs, and the model drug with a lower water solubility tended to exhibit a greater extent of over-dissolution.

    添付ファイル: P0SF-DrugProof.pdf

    DOI: 10.1021/acs.molpharmaceut.6b01048

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  • Adsorption of lysozyme on base metal surfaces in the presence of an external electric potential 査読

    Ei Ei Htwe, Yuhi Nakama, Hiroshi Tanaka, Hiroyuki Imanaka, Naoyuki Ishida, Koreyoshi Imamura

    COLLOIDS AND SURFACES B-BIOINTERFACES   147   9 - 16   2016年11月

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    担当区分:最終著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE BV  

    The impact of external electric potential on the adsorption of a protein to base metal surfaces was examined. Hen egg white lysozyme (LSZ) and six types of base metal plates (stainless steel SUS316L (St), Ti, Ta, Zr, Cr, or Ni) were used as the protein and adsorption surface, respectively. LSZ was allowed to adsorb on the surface under different conditions (surface potential, pH, electrolyte type and concentration, surface material), which was monitored using an ellipsometer. LSZ adsorption was minimized in the potential range above a certain threshold and, in the surface potential range below the threshold, decreasing the surface potential increased the amount of protein adsorbed. The threshold potential for LSZ adsorption was shifted toward a positive value with increasing pH and was lower for Ta and Zr than for the others. A divalent anion salt (K2SO4) as an electrolyte exhibited the adsorption of LSZ in the positive potential range while a monovalent salt (KCI) did not. A comprehensive consideration of the obtained results suggests that two modes of interactions, namely the electric force by an external electric field and electrostatic interactions with ionized surface hydroxyl groups, act on the LSZ molecules and determine the extent of suppression of LSZ adsorption, All these findings appear to support the view that a base metal surface can be controlled for the affinity to a protein by manipulating the surface electric potential as has been reported on some electrode materials. (C) 2016 Elsevier B.V. All rights reserved.

    添付ファイル: P2LSZ-pub.pdf

    DOI: 10.1016/j.colsurfb.2016.07.042

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  • Molecular design of proteinaceous cushion for sensitive biomolecular interaction detection system

    Hiroyuki Imanaka, Koki Dare, Koreyoshi Imamura

    NEW BIOTECHNOLOGY   33   S72 - S72   2016年7月

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    担当区分:最終著者   記述言語:英語   出版者・発行元:ELSEVIER SCIENCE BV  

    DOI: 10.1016/j.nbt.2016.06.966

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  • Surfactant-free solid dispersion of fat-soluble flavour in an amorphous sugar matrix 査読

    Tomo Satoh, Fumihiro Hidaka, Kento Miyake, Natsuki Yoshiyama, Koji Takeda, Tsutashi Matsuura, Hiroyuki Imanaka, Naoyuki Ishida, Koreyoshi Imamura

    FOOD CHEMISTRY   197   1136 - 1142   2016年4月

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    担当区分:最終著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCI LTD  

    A solid dispersion technique to homogeneously disperse hydrophobic ingredients in a water-soluble solid without using surfactant was examined as follows: first, freeze-dried amorphous sugar was dissolved in an organic medium that contained a soluble model hydrophobic component. Second, the mixed solution of sugar and the model hydrophobic component was vacuum dried into a solid (solid dispersion). Methanol and six fat-soluble flavours, including cinnamaldehyde, were used as organic media and model hydrophobic components. The retention of flavours in the solid dispersion during drying and storage under vacuum was evaluated. The amorphised disaccharides dissolved in methanol up to 100 mg/mL, even temporarily (20 s to 10 days) and could be solidified without any evidence of crystallisation and segregation from flavour. The solid dispersion, prepared using a-maltose usually showed 65-95% flavour retention during drying (and storage for cinnamaldehyde), whereas &gt;= 50% of the flavour was lost when the flavour was O/W emulsified with a surfactant and then freeze-dried with sugar. (C) 2015 Elsevier Ltd. All rights reserved.

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  • The use of a proteinaceous "cushion" with a polystyrene-binding peptide tag to control the orientation and function of a target peptide adsorbed to a hydrophilic polystyrene surface 査読

    Hiroyuki Imanaka, Daisuke Yamadzumi, Keisuke Yanagita, Naoyuki Ishida, Kazuhiro Nakanishi, Koreyoshi Imamura

    BIOTECHNOLOGY PROGRESS   32 ( 2 )   527 - 534   2016年3月

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    担当区分:最終著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-BLACKWELL  

    In immobilizing target biomolecules on a solid surface, it is essential (i) to orient the target moiety in a preferred direction and (ii) to avoid unwanted interactions of the target moiety including with the solid surface. The preferred orientation of the target moiety can be achieved by genetic conjugation of an affinity peptide tag specific to the immobilization surface. Herein, we report on a strategy for reducing the extent of direct interaction between the target moiety and surface in the immobilization of hexahistidine peptide (6His) and green fluorescent protein (GFP) on a hydrophilic polystyrene (PS) surface: Ribonuclease HII from Thermococcus kodakaraensis (cHII) was genetically inserted as a cushion between the PS-affinity peptide tag and target moiety. The insertion of a cushion protein resulted in a considerably stronger immobilization of target biomolecules compared to conjugation with only a PS affinity peptide tag, resulting in a substantially enhanced accessibility of the detection antibody to the target 6His peptide. The fluorescent intensity of the GFP moiety was decreased by approximately 30% as the result of fusion with cHII and the PS-affinity peptide tag but was fully retained in the immobilization on the PS surface irrespective of the increased binding force. Furthermore, the fusion of cHII did not impair the stability of the target GFP moiety. Accordingly, the use of a proteinaceous cushion appears to be promising for the immobilization of functional biomolecules on a solid surface. (c) 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:527-534, 2016

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  • クッションタンパク質の分子デザインによるバイオ分子間相互作用の高感度化

    今中 洋行, 伊達 弘輝, 的場 晴香, 石田 尚之, 今村 維克

    表面科学学術講演会要旨集   36   37 - 37   2016年

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    記述言語:日本語   出版者・発行元:公益社団法人 日本表面科学会  

    ELISA,SPRやQCMなどのバイオ分子間相互作用検出系における基板上へのリガンド分子の固定化について,基板親和性ペプチド連結による固定化配向制御ならびにリガンド分子と基板との直接的な相互作用を緩和するクッションタンパク質を用いた機能維持を試みた.そして,クッションタンパク質の様々な分子デザインにより,ペプチド-タンパク質間あるいはタンパク質-タンパク質間相互作用検出を顕著に高感度化できた.

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  • Effect of surface hydrophobicity on short-range hydrophobic attraction between silanated silica surfaces 査読

    Yuhei Soga, Hiroyuki Imanaka, Koreyoshi Imamura, Naoyuki Ishida

    ADVANCED POWDER TECHNOLOGY   26 ( 6 )   1729 - 1733   2015年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE BV  

    The interaction forces between silanated silica surfaces without nanobubbles were measured using colloidal probe atomic force microscopy (AFM). To obtain hydrophobic surfaces without nanobubbles, an aqueous solution was introduced between the surfaces following an exchange process involving several solvents. In the obtained approaching force curves, an attractive force was observed from a distance of 10-20 nm, which is an additional attractive force stronger than typical van der Waals attractions. When the surface hydrophobicity decreased, the range of this attraction decreased slightly; the attraction disappeared when the surface contact angle was below 90 degrees. In contrast, measurements in the water-ethanol mixtures revealed that the attraction persisted even when the contact angle was well below 90 degrees. The possible origin of this force was discussed on the basis of the obtained results. (C) 2015 The Society of Powder Technology Japan. Published by Elsevier B.V. and The Society of Powder Technology Japan.

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  • Inhibitory effects of additives and heat treatment on the crystallization of freeze-dried sugar 査読

    Kohshi Kinugawa, Mitsunori Kinuhata, Ryo Kagotani, Hiroyuki Imanaka, Naoyuki Ishida, Mizuki Kitamatsu, Kazuhiro Nakanishi, Koreyoshi Imamura

    JOURNAL OF FOOD ENGINEERING   155   37 - 44   2015年6月

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    担当区分:最終著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCI LTD  

    An amorphous matrix of a sugar is frequently used as a bulk-forming and stabilizing agent in the food industry but tends to crystallize as the result of water uptake and increase in temperature. Additives and methods used to inhibit the crystallization of amorphous sugar (sucrose) were screened in this study. Freeze-dried amorphous sucrose containing 0.5-5 wt% of additive, including salts, different types of sugars, and polymers, the crystallization temperature (T-cry) and isothermal crystallization characteristics were examined. Certain types of salts markedly increased the Tcry and prolonged the induction period for crystal nucleation. The use of 1 wt% MgCl2 was particularly effective in inhibiting sugar crystallization. The heat treatment of crystalline sucrose under appropriate conditions was also found to result in diminished sucrose crystallization. MALDI-TOF mass spectra of the heat-treated sucrose suggested that sucrose derivatives containing multiple pyranose groups were formed, which would closely relate to the crystallization inhibition. Finally, the protein stabilizing effects of the matrices were evaluated. The results indicated that both the addition of additives and the heat treatment resulted in an improvement of the protein stabilizing effect of amorphous sugar matrix, compared to that of sucrose alone. (C) 2015 Elsevier Ltd. All rights reserved.

    添付ファイル: P0CryInh-proof.pdf

    DOI: 10.1016/j.jfoodeng.2015.01.016

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  • Influence of sugar surfactant structure on the encapsulation of oil droplets in an amorphous sugar matrix during freeze-drying 査読

    Shota Nakayama, Yoshifumi Kimura, Sayuri Miki, Jun Oshitani, Takashi Kobayashi, Shuji Adachi, Tsutashi Matsuura, Hiroyuki Imanaka, Naoyuki Ishida, Hiroko Tada, Kazuhiro Nakanishi, Koreyoshi Imamura

    FOOD RESEARCH INTERNATIONAL   70   143 - 149   2015年4月

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    担当区分:最終著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE BV  

    The encapsulation of O/W emulsion droplets in a freeze-dried amorphous sugar matrix was investigated, focusing on the impact of the molecular structure of the emulsifying surfactant. O/W emulsions, containing various surfactants, were freeze-dried in the presence of a sugar. Thirty types of surfactants, including eighteen different sugar surfactants and ten types of commercially available sugar ester mixtures, were used. Linoleic acid methyl ester and trehalose were used as the oil phase and sugar. The amounts of oil droplets encapsulated in freeze-dried amorphous sugar matrix were analyzed by Fourier transform Infrared spectroscopy. Sugar surfactants were generally superior to the other classes of surfactants for oil droplet encapsulation during freeze-drying, and there was the optimum alkyl chain length of the sugar surfactant. Sugar esters generally exhibited greater oil encapsulation than sugar ethers. Larger sugar head group appeared to result in better encapsulation in the case of sugar esters, but the opposite tendency was found for sugar ethers. A limited combination of sugar surfactants (15% sucrose mono- and 85% di-stearate) resulted in the maximum oil droplet encapsulation efficiency although these surfactants are individually quite poor in the encapsulation and other tested combinations did not improve the encapsulation efficiency relative to their individual effectiveness. (C) 2015 Elsevier Ltd. All rights reserved.

    添付ファイル: P1AOex-pub.pdf

    DOI: 10.1016/j.foodres.2015.02.003

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  • Effect of Lipid Amount on Surfactant-free Solid Lipid Nanoparticle Formation by Hot Homogenization 査読

    Jun Oshitani, Natsumi Murakami, Mikio Yoshida, Naoyuki Ishida, Koreyoshi Imamura, Hideki Ichikawa

    CHEMISTRY LETTERS   43 ( 7 )   1011 - 1013   2014年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:CHEMICAL SOC JAPAN  

    The yield and size distribution of surfactant-free solid lipid nanoparticles (SF-SLNs) formed by hot homogenization were investigated by changing the amount of lipid (palmitic acid). The yield was 100% and monodispersed same-sized particles of 120-200 nm were formed at the lipid amount &lt;= 2.8 x 10(-4) g-lipid (g-water)(-1). The yield decreased owing to phase separation, and the size increased "step-likely" to be 200 350 nm while keeping the monodispersity when the lipid amount was just above &lt;= 2.8 x 10(-4) g-lipid (g-water)(-1). The size and the lipid amount continued to increase gradually at the lipid amount &gt;2.8 x 10(-4) g-lipid (g-water)(-1). The results indicate that submicron-sized solid lipid particles can be formed without surfactants and that the decrease of yield and the step-like increase of particle size occur at the same lipid amount. The total surface area of the SF-SLNs was estimated using the experimental data. It is suggested that the total surface area and the lipid amount are correlated to each other.

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  • Characteristics of Sugar Surfactants in Stabilizing Proteins During Freeze-Thawing and Freeze-Drying 査読

    Koreyoshi Imamura, Katsuyuki Murai, Tamayo Korehisa, Noriyuki Shimizu, Ryo Yamahira, Tsutashi Matsuura, Hiroko Tada, Hiroyuki Imanaka, Naoyuki Ishida, Kazuhiro Nakanishi

    JOURNAL OF PHARMACEUTICAL SCIENCES   103 ( 6 )   1628 - 1637   2014年6月

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    担当区分:筆頭著者, 責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-BLACKWELL  

    Sugar surfactants with different alkyl chain lengths and sugar head groups were compared for their protein-stabilizing effect during freeze-thawing and freeze-drying. Six enzymes, different in terms of tolerance against inactivation because of freeze-thawing and freeze-drying, were used as model proteins. The enzyme activities that remained after freeze-thawing and freeze-drying in the presence of a sugar surfactant were measured for different types and concentrations of sugar surfactants. Sugar surfactants stabilized all of the tested enzymes both during freeze-thawing and freeze-drying, and a one or two order higher amount of added sugar surfactant was required for achieving protein stabilization during freeze-drying than for the cryoprotection. The comprehensive comparison showed that the C10-C12 esters of sucrose or trehalose were the most effective through the freeze-drying process: the remaining enzyme activities after freeze-thawing and freeze-drying increased at the sugar ester concentrations of 1-10 and 10-100 M, respectively, and increased to a greater extent than for the other surfactants at higher concentrations. Results also indicate that, when a decent amount of sugar was also added, the protein-stabilizing effect of a small amount of sugar ester through the freeze-drying process could be enhanced. (c) 2014 Wiley Periodicals, Inc. and the American Pharmacists Association J Pharm Sci

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  • 糖界面活性剤を用いたタンパク質の安定化

    今村 維克

    オレオサイエンス   14 ( 11 )   487 - 493   2014年

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    記述言語:日本語   出版者・発行元:公益社団法人 日本油化学会  

    <p>タンパク質は食品や医薬品分野で不可欠な材料であるが,非生理的環境下では容易に変性し,その優れた生理活性を失ってしまう場合が多い。そのためタンパク質の安定化技術,とりわけ安定化物質の探索が以前より行われてきた。一方,糖を親水基とする糖界面活性剤は食品添加物として広範に利用されているが,タンパク質の変性を抑制する作用も有している。本稿では,タンパク質が曝される各状態・操作における変性機構とともに,糖界面活性剤のタンパク質安定化における有効性について述べる。特に凍結および凍結乾燥過程におけるタンパク質安定化作用について,種々の糖界面活性剤が各種酵素の活性保持に対する影響を調査し,糖界面活性剤の極性基の構造やアルキル鎖長,およびアルキル鎖との結合形態が凍結および凍結乾燥時におけるタンパク質安定化作用に及ぼす影響について考察を加えた。</p>

    DOI: 10.5650/oleoscience.14.487

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    その他リンク: http://search.jamas.or.jp/link/ui/2015030450

  • 水中の疎水化シリカ表面間に働く短距離性疎水性引力に及ぼす表面の疎水性の影響

    曽我 友平, 今中 洋行, 今村 維克, 石田 尚之

    粉体工学会誌   51 ( 5 )   343 - 348   2014年

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    記述言語:英語   出版者・発行元:一般社団法人 粉体工学会  

    The interaction forces between silanated silica surfaces with no nanobubbles were measured using colloidal probe atomic force microscopy (AFM). To obtain hydrophobic surfaces without nanobubbles, an aqueous solution was introduced between the surfaces following an exchange process involving several solvents. In the obtained approachingforce curves, an attractive force was observed from a distance of 10~20 nm, which is an additional attractive force stronger than typical van der Waals attractions. When the surface hydrophobicity decreased, the range of this attraction decreased slightly ; the attraction disappeared when the surface contact angle was below 90°。. In contrast, measurements in the water-ethanol mixtures revealed that the attraction persisted even when the contact angle was well below 90°。. The possible origin of this force was discussed on the basis of the obtained results.

    DOI: 10.4164/sptj.51.343

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  • 2P078 新規金表面親和性ペプチドの同定及びタンパク質固定化への応用(01D. 蛋白質:機能,ポスター,第52回日本生物物理学会年会(2014年度))

    Shigemori Yojiro, Yoshida Kaori, Imamura Koreyoshi, Takahashi Yuichiro, Imanaka Hiroyuki

    生物物理   54 ( 1 )   S207   2014年

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    記述言語:英語   出版者・発行元:一般社団法人 日本生物物理学会  

    DOI: 10.2142/biophys.54.S207_6

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  • 2P085 CS複合体形成をモデルとした高感度ペプチド-タンパク質間相互作用検出系のデザイン(01F. 蛋白質:蛋白質工学/進化工学,ポスター,第52回日本生物物理学会年会(2014年度))

    Matsushita Runa, Ishida Naoyuki, Imamura Koreyoshi, Imanaka Hiroyuki

    生物物理   54 ( 1 )   S209   2014年

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    記述言語:英語   出版者・発行元:一般社団法人 日本生物物理学会  

    DOI: 10.2142/biophys.54.S209_1

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  • Improving the physical stability of freeze-dried amorphous sugar matrices by compression at several hundreds MPa 査読

    Ryo Kagotani, Kohshi Kinugawa, Mayo Nomura, Hiroyuki Imanaka, Naoyuki Ishida, Koreyoshi Imamura

    JOURNAL OF PHARMACEUTICAL SCIENCES   102 ( 7 )   2187 - 2197   2013年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-BLACKWELL  

    Amorphous matrices, composed of sugars, are markedly plasticized by moisture uptake, which results in physical instability. Our previous studies, in the compression pressure range 443 MPa, indicated that when a matrix is compressed, the amount of sorbed water at given relative humidities (RHs) decreases, whereas the glass transition temperature (Tg) remains constant. Herein, the effect of higher compression pressures than those used previously was explored to investigate the feasibility of using compression to improve the physical stability of amorphous sugar matrix against water uptake and subsequent collapse. Amorphous sugar samples were prepared by freeze-drying and then compressed at 0-665 MPa, followed by rehumidification at given RHs. The physical stability of the amorphous sugar sample was evaluated by measuring Tg and crystallization temperature (Tcry). The amounts of sorbed water, different in the interaction state, were determined using an FTIR technique. It was found that the compression at pressures of 443 MPa decreased the amount of sorbed water, which is a major factor in plasticization and crystallization, and thus markedly increased the Tg and Tcry relative to that for the uncompressed sample. Hence, the compression at several hundreds MPa appears to be feasible for improving the physical stability of amorphous sugar matrix. (c) 2013 Wiley Periodicals, Inc. and the American Pharmacists Association J Pharm Sci 102:2187-2197, 2013

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  • Improvement on the freeze-thaw stability of corn starch gel by the polysaccharide from leaves of Corchorus olitorius L. 査読

    Eiji Yamazaki, Toru Sago, Yoshiaki Kasubuchi, Kazuhito Imamura, Toshio Matsuoka, Osamu Kurita, Hironobu Nambu, Yasuki Matsumura

    Carbohydrate Polymers   94 ( 1 )   555 - 560   2013年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Effect of the polysaccharide from leaves of Corchorus olitorius L. (PLC) on the freeze-thaw (FT) stability of corn starch gel was studied. PLC was incorporated into the starch gel at 0.7% and total solid was adjusted to 6.0%. The syneresis was measured by the centrifugal-filtration method and, as a result, addition of PLC reduced effectively the syneresis of the starch gel even after 5 FT cycles, which was less than one third that of the normal starch gel. The rheological changes of the starch/PLC gel during the FT treatments were evaluated while the gel remained on the rheometer plate. The starch/PLC gel had less significant changes in the rheological parameters during the FT cycles than starch/guar gum or xanthan gum gel systems. SEM images showed that PLC stabilized the gel matrix surrounding pores, which would contribute to both a lower syneresis production and a higher stability in the rheological behavior at FT. © 2013 Elsevier Ltd. All rights reserved.

    DOI: 10.1016/j.carbpol.2013.01.079

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  • Characteristics of amorphous matrices composed of different types of sugars in encapsulating emulsion oil droplets during freeze-drying 査読

    Koreyoshi Imamura, Yoshifumi Kimura, Shota Nakayama, Miki Sayuri, Seiji Ogawa, Tatsuya Hoshino, Jun Oshitani, Takashi Kobayashi, Shuji Adachi, Tsutashi Matsuura, Hiroyuki Imanaka, Naoyuki Ishida, Kazuhiro Nakanishi

    FOOD RESEARCH INTERNATIONAL   51 ( 1 )   201 - 207   2013年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE BV  

    The encapsulation of emulsion oil droplets by amorphous sugar matrices, formed by freeze-drying, was investigated, with a focus on the influence of the type of sugar. An oil-in-water emulsion, comprised of linoleic acid methyl ester (LME) and sucrose monolaurate (SML) as an oil phase and surfactant, respectively, were freeze-dried in the presence of different types of sugars. LME-droplet encapsulation during and after freeze-drying were evaluated by FTIR analysis. The loss of LME largely occurred in the early stage of freeze-drying. The size distribution of the encapsulated LME droplets remained unchanged before and after freeze-drying in most cases. The encapsulated fractions of LME droplets could be correlated with the glass transition temperature of the sugars in the fully hydrated state (T-g*), and the existence of an optimum T-g* value for the sugar matrix was predicted. The encapsulation ability of an amorphous sugar matrix was maximized when mono- and polysaccharide were combined so as to give a value for T-g* of approximately -50 degrees C, although, individually, mono- and polysaccharides were quite poor for oil droplet encapsulation. These findings suggest that the structural flexibility of the amorphous sugar matrix is a major determinant in oil droplet encapsulation by an amorphous sugar matrix during freeze-drying. (C) 2012 Elsevier Ltd. All rights reserved.

    DOI: 10.1016/j.foodres.2012.12.010

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  • On the preparation of indoxyl red from indican and some new characteristics 査読

    Jingyuan Song, Mizuki Kitamatsu, Koreyoshi Imamura, Hitoshi Ohmori, Kouji Watanabe, Kazuhiro Nakanishi

    BIOORGANIC & MEDICINAL CHEMISTRY LETTERS   23 ( 3 )   627 - 629   2013年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:PERGAMON-ELSEVIER SCIENCE LTD  

    An indole compound with a strong purple-red color was produced by boiling a solution of indican under acidic conditions and purified by chromatographies on DEAE-650S Toyopearl TSK-gel and silica-gel columns. The purple-red compound purified was identified as indoxyl red, on the basis of FAB Mass, C-13 NMR, H-1 NMR, UV-visible spectra, and IR spectra. Although indoxyl red was first synthesized by Seidel(9) 70 years ago, very little information has been available on its characteristics. We repot here that the compound was purple-red colored at acidic pH and green at pH 13, and showed antiproliferative and cytotoxic activities to the mouse B cell lymphoma cell line NSF202. (c) 2012 Elsevier Ltd. All rights reserved.

    DOI: 10.1016/j.bmcl.2012.12.006

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  • The discoidin domain of Bacillus circulans β-galactosidase plays an essential role in repressing galactooligosaccharide production. 査読

    Song J, Imanaka H, Imamura K, Minoda M, Yamaguchi S, Nakanishi K

    Bioscience, biotechnology, and biochemistry   77 ( 1 )   73 - 79   2013年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1271/bbb.120583

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  • Impact of compression, physical aging, and freezing rate on the crystallization characteristics of an amorphous sugar matrix 査読

    Koreyoshi Imamura, Kohshi Kinugawa, Ryo Kagotani, Mayo Nomura, Kazuhiro Nakanishi

    JOURNAL OF FOOD ENGINEERING   112 ( 4 )   313 - 318   2012年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCI LTD  

    Amorphous matrices made up of sugar molecules, are frequently used in food and pharmaceutical industries. A drawback to their use is that they are susceptible to collapse, as a result of water uptake and an increase in temperature and subsequently crystallize. Herein, the crystallization characteristics of amorphous sugar (sucrose and alpha-lactose) preparations were analyzed, with the purpose of obtaining knowledge that could lead to the prediction of how long the amorphous state is retained under various conditions. The impact of compression, physical aging and freezing rate on the induction period (t(ind)) for crystallization were examined. Freeze-dried sugar samples were compressed at 74 or 443 MPa (5 min) and then rehumidified at specified RHs. Some freeze-dried sucrose samples were physically aged, and alternatively freeze-drying was conducted under different conditions. The isothermal crystallization of the prepared samples at different temperatures (T), the glass transition and the crystallization temperature (T-cry) were measured, using differential scanning calorimetry. The compression markedly decreased the t(ind), while significantly lowered the hygroscopicity. Physical aging and slower-freezing also shortened the t(ind). The t(ind) was found to be correlated exclusively with (T-cry-T), regardless of rehumidification, compression, sugar type, physical aging and freezing rate in the freeze-drying process. (C) 2012 Elsevier Ltd. All rights reserved.

    DOI: 10.1016/j.jfoodeng.2012.05.004

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  • Heterogeneity of the state and functionality of water molecules sorbed in an amorphous sugar matrix 査読

    Koreyoshi Imamura, Ryo Kagotani, Mayo Nomura, Kohshi Kinugawa, Kazuhiro Nakanishi

    CARBOHYDRATE RESEARCH   351   108 - 113   2012年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCI LTD  

    An amorphous matrix, comprised of sugar molecules, is frequently used in the pharmaceutical industry. An amorphous sugar matrix exhibits high hygroscopicity, and it has been established that the sorbed water lowers the glass transition temperature T-g of the amorphous sugar matrix. It is naturally expected that the random allocation and configuration of sugar molecules would result in heterogeneity of states for sorbed water. However, most analyses of the behavior of water, when sorbed to an amorphous sugar matrix, have implicitly assumed that all of the sorbed water molecules are in a single state. In this study, the states of water molecules sorbed in an amorphous sugar matrix were analyzed by Fourier-transform IR spectroscopy and a Fourier self-deconvolution technique. When sorbed water molecules were classified into five states, according to the extent to which they are restricted, three of the states resulted in a lowering of T-g of an amorphous sugar matrix, while the other two were independent of the plasticization of the matrix. This finding provides an explanation for the paradoxical fact that compression at several hundreds of MPa significantly decreases the equilibrium water content at a given RH, while the T-g remains unchanged. (C) 2011 Elsevier Ltd. All rights reserved.

    DOI: 10.1016/j.carres.2011.12.021

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  • Rod-shaped Hydroxyapatite Nanoparticle Formation by Maturation with AOT 査読

    Jun Oshitani, Nobuhide Kataoka, Mikio Yoshida, Kuniaki Gotoh, Koreyoshi Imamura

    CHEMISTRY LETTERS   40 ( 10 )   1085 - 1086   2011年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:CHEMICAL SOC JAPAN  

    Amorphous calcium phosphate nanoparticles were synthesized by mixing aqueous solutions of Ca(NO3)(2) and (NH4)(2)-HPO4. Then the particles were matured with sodium bis-(2-ethylhexyl) sulfosuccinate (AOT) at various temperatures. Rod-shaped hydroxyapatite (HAp) nanoparticles were formed at maturation temperature &gt;= 40 degrees C. These results indicate that addition of AOT nor during the particles synthesis but during the maturation at more equal critical temperature is a key to form the rod-shaped HAp nanoparticles using AOT.

    DOI: 10.1246/cl.2011.1085

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  • Cloning and expression of a β-galactosidase gene of Bacillus circulans. 査読

    Song J, Imanaka H, Imamura K, Minoda M, Katase T, Hoshi Y, Yamaguchi S, Nakanishi K

    Bioscience, biotechnology, and biochemistry   75 ( 6 )   1194 - 1197   2011年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1271/bbb.110014

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  • Rod-shaped Hydroxyapatite Nanoparticle Formation Using AOT and Water without Oil 査読

    Jun Oshitani, Nobuhide Kataoka, Mikio Yoshida, Kuniaki Gotoh, Koreyoshi Imamura, Hidekazu Tanaka

    CHEMISTRY LETTERS   40 ( 4 )   400 - 401   2011年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:CHEMICAL SOC JAPAN  

    We synthesized rod-shaped hydroxyapatite (HAp) nanoparticles using sodium bis(2-ethylhexyl) sulfosuccinate (AOT) as an additive in water without an oil. The length of the particles increases from 40 to 100 nm with increasing the concentration ratio AOT/Ca while maintaining a width of 12 nm. These results indicate that an oil phase is not necessarily needed to form the rod-shaped HAp nanoparticles using AOT.

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  • Influence of compression on water sorption, glass transition, and enthalpy relaxation behavior of freeze-dried amorphous sugar matrices 査読

    Koreyoshi Imamura, Ryo Kagotani, Mayo Nomura, Kazuhiro Tanaka, Kohshi Kinugawa, Kazuhiro Nakanishi

    INTERNATIONAL JOURNAL OF PHARMACEUTICS   408 ( 1-2 )   76 - 83   2011年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE BV  

    An amorphous matrix comprised of sugar molecules are frequently used in the pharmaceutical industry. The compression of the amorphous sugar matrix improves the handling. Herein, the influence of compression on the water sorption of an amorphous sugar matrix was investigated. Amorphous sugar samples were prepared by freeze-drying, using several types of sugars, and compressed at 0-443 MPa. The compressed amorphous sugar samples as well as uncompressed samples were rehumidified at given RHs, and the equilibrium water content and glass transition temperature (T-g) were then measured. Compression resulted in a decrease in the equilibrium water content of the matrix, the magnitude of which was more significant for smaller sized sugars. Diffusivity of water vapor in the sample was also decreased to one-hundredth by the compression. The T-g value for a given RH remained unchanged, irrespective of the compression. Accordingly, the decrease in T-g with increasing water content increased as the result of compression. The structural relaxation of the amorphous sugar matrices were also examined and found to be accelerated to the level of a non-porous amorphous sugar matrix as the result of the compression. The findings indicate that pores contained in freeze-dried sugar samples interfere with the propagation of structural relaxation. (C) 2011 Elsevier B.V. All rights reserved.

    DOI: 10.1016/j.ijpharm.2011.01.052

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  • Causes of the production of multiple forms of β-galactosidase by Bacillus circulans. 査読

    Song J, Abe K, Imanaka H, Imamura K, Minoda M, Yamaguchi S, Nakanishi K

    Bioscience, biotechnology, and biochemistry   75 ( 2 )   268 - 278   2011年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1271/bbb.100574

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  • Multiple States of Sorbed Water Molecules in Amorphous Sugar Matrix andtheir Different Impacts on Physical Properties of the Matrix 査読

    Koreyoshi Imamura

    japan journal of food engineering   12 ( 1 )   1 - 9   2011年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Amorphous materials, comprised of sugar molecules, exhibit high hygroscopicity and the sorbed water exerts a major influence on physical stability of the matrix. To date, the water sorption behavior of amorphous sugars has been extensively investigated. However, most studies of the behavior of water, when sorbed to an amorphous sugar matrix, have implicitly assumed that all of the sorbed water molecules are in a single state: It is naturally expected that the random allocation and configuration of sugar molecules would result in heterogeneity of states for the sorbed water. Recently, we demonstrated the heterogeneity of state and functionality of water sorbed in amorphous sugar matrices by combining a Fourier transform IR spectroscopy and Fourier self-deconvolution technique
    The sorbed water molecules were classified into five states, and the three of them mainly served to lower the glass transition temperature of an amorphous sugar matrix while the other two appeared to be independent of physical properties of the matrix. © 2011, Japan Society for Food Engineering. All rights reserved.

    DOI: 10.11301/jsfe.12.1

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  • Water Sorption, Glass Transition, and Protein-Stabilizing Behavior of an Amorphous Sucrose Matrix Combined With Various Materials 査読

    Koreyoshi Imamura, Toru Yokoyama, Atsushi Fukushima, Mitsunori Kinuhata, Kazuhiro Nakanishi

    JOURNAL OF PHARMACEUTICAL SCIENCES   99 ( 11 )   4669 - 4677   2010年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:JOHN WILEY & SONS INC  

    The effects of various additives on the physical properties of an amorphous sugar matrix were compared. Amorphous, sugar-additive mixtures were prepared by freeze-drying and then rehumidified at given RHs. Sucrose and eighteen types of substances were used as the sugar and the additive, respectively, and water sorption, glass-to-rubber transition, and protein stabilization during freeze-drying for the various sucrose-additive mixtures were examined. The additives were categorized into two groups according to their effects on T(g) and water sorption. Presence of polysaccharides, cyclodextrins, and polymers (large-sized additives) resulted in a decrease in equilibrium water content from the ideal value calculated from individual water contents for sucrose and additive, and in contrast, low MW substances containing ionizable groups (small-ionized additives) resulted in an increase. The increase in T(g) by the addition of large-sized additives was significant at the additive contents &gt;50 wt.% whereas the T(g) was markedly increased in the lower additive content by the addition of small-ionized additives. The addition of small-ionized additives enhanced the decrease in T(g) with increasing water content. The protein stabilizing effect was decreased with increasing additive content in the cases of the both groups of the additives. (C) 2010 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 99:4669-4677, 2010

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  • Development of a highly efficient indigo dyeing method using indican with an immobilized beta-glucosidase from Aspergillus niger 査読

    Jingyuan Song, Hiroyuki Imanaka, Koreyoshi Imamura, Kouichi Kajitani, Kazuhiro Nakanishi

    JOURNAL OF BIOSCIENCE AND BIOENGINEERING   110 ( 3 )   281 - 287   2010年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:SOC BIOSCIENCE BIOENGINEERING JAPAN  

    A highly efficient method for dyeing textiles with indigo is described. In this method, the substrate, indican is first hydrolyzed at an acidic pH of 3 using an immobilized beta-glucosidase to produce indoxyl, under which conditions indigo formation is substantially repressed. The textile sample is then dipped in the prepared indoxyl solution and the textile is finally exposed to ammonia vapor for a short time, resulting in rapid indigo dyeing. As an enzyme, we selected a beta-glucosidase from Aspergillus niger, which shows a high hydrolytic activity towards indican and was thermally stable at temperatures up to 50-60 degrees C, in an acidic pH region. The A. niger beta-glucosidase, when immobilized on Chitopearl BCW-3001 by treatment with glutaraldehyde, showed an optimum reaction pH similar to that of the free enzyme with a slightly higher thermal stability. The kinetics for the hydrolysis of indican at pH 3, using the purified free and immobilized enzymes was found to follow Michaelis-Menten type kinetics with weak competitive inhibition by glucose. Using the immobilized enzyme, we successfully carried out repeated-batch and continuous hydrolyses of indican at pH 3 when nitrogen gas was continuously supplied to the substrate solution. Various types of model textiles were dyed using the proposed method although the color yield varied, depending on the type of textile used. (C) 2010, The Society for Biotechnology, Japan. All rights reserved.

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  • Influences of properties of protein and adsorption surface on removal kinetics of protein adsorbed on metal surface by H2O2-electrolysis treatment 査読

    Koreyoshi Imamura, Manabu Oshita, Masumi Iwai, Toshiki Kuroda, Ippei Watanabe, Takaharu Sakiyama, Kazuhiro Nakanishi

    JOURNAL OF COLLOID AND INTERFACE SCIENCE   345 ( 2 )   474 - 480   2010年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ACADEMIC PRESS INC ELSEVIER SCIENCE  

    "H2O2-electrolysis" treatment is an alternative method for removing proteinaceous materials that are adsorbed to metal surfaces. The method is based on the generation of hydroxyl radicals by electrolysis of hydrogen peroxide and the subsequent decomposition of organic substances adhering to the metal surface. We herein investigated the influence of some parameters on the kinetics of protein removal by H2O2-electrolysis. These parameters included the properties of proteins and the type of metal surface. Sixteen types of proteins and nine types of metal surfaces were used. The removal of adsorbed protein from a metal surface during the treatment was monitored by ellipsometry. Apparent first-order rate constants for removal, k(cl), for various adsorption and treatment conditions were determined. The k(cl) value varied markedly with the type of protein and was also influenced by the pH used in the adsorption. The isoelectric point (pI) of protein used was found to be a major factor. The amount of adsorbed protein removed by a unit amount of (OH)-O-center dot was larger for a metal surface with a lower pI. The impact of the properties of the protein and metal surface on the removal kinetics are discussed, focusing on relationships with the adsorption characteristics of the protein. (C) 2010 Elsevier Inc. All rights reserved.

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  • Cultivation characteristics and gene expression profiles of Aspergillus oryzae by membrane-surface liquid culture, shaking-flask culture, and agar-plate culture 査読

    Hiroyuki Imanaka, Soukichi Tanaka, Bin Feng, Koreyoshi Imamura, Kazuhiro Nakanishi

    JOURNAL OF BIOSCIENCE AND BIOENGINEERING   109 ( 3 )   267 - 273   2010年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:SOC BIOSCIENCE BIOENGINEERING JAPAN  

    We cultivated a filamentous fungus, Aspergillus oryzae IAM 2706 by three different cultivation methods, i.e., shaking-flask culture (SFC), agar-plate culture (APC), and membrane-surface liquid culture (MSLC), to elucidate the differences of its behaviors by different cultivation methods under the same media, by measuring the growth, secretion of proteases and alpha-amylase, secreted protein level, and gene transcriptional profile by the DNA microarray analysis. The protease activities detected by MSLC and APC were much higher than that by SFC, using both modified Czapek-Dox (mCD) and dextrin-peptone-yeast extract (DPY) media. The alpha-amylase activity was detected in MSLC and APC in a much larger extent than that in SFC when DPY medium was used. On the basis of SDS-PAGE analyses and N-terminal amino acid sequences, 6 proteins were identified in the supernatants of the culture broths using DPY medium, among which oryzin (alkaline protease) and alpha-amylase were detected at a much higher extent for APC and MSLC than those for SFC while only oryzin was detected in mCD medium, in accordance with the activity measurements. A microarray analysis for the fungi cultivated by SFC, APC, and MSLC using mCD medium was carried out to elucidate the differences in the gene transcriptional profile by the cultivation methods. The gene transcriptional profile obtained for the MSLC sample showed a similar tendency to the APC sample while it was quite different from that for the SFC sample. Most of the genes specifically transcribed in the MSLC sample versus those in the SFC sample with a 10-fold up-regulation or higher were unknown or predicted proteins. However, transcription of oryzin gene was only slightly up-regulated in the MSLC sample and that of alpha-amylase gene, slightly down-regulated. (C) 2009, The Society for Biotechnology, Japan. All rights reserved.

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  • Impacts of Compression on Crystallization Behavior of Freeze-Dried Amorphous Sucrose 査読

    Koreyoshi Imamura, Mayo Nomura, Kazuhiro Tanaka, Nobuhide Kataoka, Jun Oshitani, Hiroyuki Imanaka, Kazuhiro Nakanishi

    JOURNAL OF PHARMACEUTICAL SCIENCES   99 ( 3 )   1452 - 1463   2010年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:JOHN WILEY & SONS INC  

    An amorphous matrix comprised of sugar molecules is used as excipient and stabilizing agent for labile ingredients in the pharmaceutical industry. The amorphous sugar matrix is often compressed into a tablet form to reduce the volume and improve handling. Herein, the effect of compression on the crystallization behavior of an amorphous sucrose matrix was investigated. Amorphous sucrose samples were prepared by freeze-drying and compressed under different conditions, followed by analyses by differential scanning calorimetry, isothermal crystallization tests, X-ray powder diffractometry, Fourier transform infrared spectroscopy (FTIR), and gas pycnometry. The compressed sample had a lower crystallization temperature and a shorter induction period for isothermal crystallization, indicating that compression facilitates the formation of the critical nucleus of a sucrose crystal. Based on FTIR and molecular dynamics simulation results, the conformational distortion of sucrose molecules due to the compression appears to contribute to the increase in the free energy of the system, which leads to the facilitation of critical nucleus formation. An isothermal crystallization test indicated an increase in the growth rate of sucrose crystals by the compression. This can be attributed to the transformation of the microstructure from porous to nonporous, as the result of compression. (C) 2009 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 99:1452-1463, 2010

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  • 低温度領域における糖類アモルファスマトリクスのタンパク質安定化作用と糖分子の束縛状態

    今村 維克, 横山 徹, 大山 健一, 絹畠 光倫, 今中 洋行, 中西 一弘

    化学工学会 研究発表講演要旨集   2010   584 - 584   2010年

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    記述言語:日本語   出版者・発行元:公益社団法人 化学工学会  

    DOI: 10.11491/scej.2010f.0.584.0

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  • 各種糖類アモルファスマトリクスに収着した水の状態とガラス転移温度との関係

    籠谷 亮, 衣川 耕史, 野村 真世, 今中 洋行, 今村 維克, 中西 一弘

    化学工学会 研究発表講演要旨集   2010   583 - 583   2010年

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    記述言語:日本語   出版者・発行元:公益社団法人 化学工学会  

    DOI: 10.11491/scej.2010f.0.583.0

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  • Streptmyces mobaraensis由来epsilon-Lysine acylaseによるN-epsilon-Lauroyl-L-Lysineの効率的合成

    内田 達也, 田口 友造, 今中 洋行, 今村 維克, 是石 真友子, 中西 一弘

    化学工学会 研究発表講演要旨集   2010   1003 - 1003   2010年

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    記述言語:日本語   出版者・発行元:公益社団法人 化学工学会  

    DOI: 10.11491/scej.2010f.0.1003.0

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  • クッションタンパク質を用いた機能的ペプチド固定化法の検討

    國方 俊暢, 今中 洋行, 今村 維克, 中西 一弘

    化学工学会 研究発表講演要旨集   2010   997 - 997   2010年

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    記述言語:日本語   出版者・発行元:公益社団法人 化学工学会  

    DOI: 10.11491/scej.2010f.0.997.0

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  • Development of peptide immobilization method using PS-tagged cushion protein 査読

    Hiroyuki Imanaka, Daisuke Yamazumi, Toshinobu Kunikata, Koreyoshi Imamura, Kazuhiro Nakanishi

    JOURNAL OF BIOSCIENCE AND BIOENGINEERING   108   S161 - S161   2009年11月

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    記述言語:英語   出版者・発行元:SOC BIOSCIENCE BIOENGINEERING JAPAN  

    DOI: 10.1016/j.jbiosc.2009.08.436

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  • Formation characteristics of calcium phosphate deposits on a metal surface by H2O2-electrolysis reaction under various conditions 査読

    Kazuaki Kanamoto, Koreyoshi Imamura, Nobuhide Kataoka, Jun Oshitani, Hiroyuki Imanaka, Kazuhiro Nakanishi

    COLLOIDS AND SURFACES A-PHYSICOCHEMICAL AND ENGINEERING ASPECTS   350 ( 1-3 )   79 - 86   2009年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE BV  

    The cathodic electrolysis of H2O2 (H2O2 + e(-) -&gt; OH- + (OH)-O-center dot) on a metal surface in the presence of calcium and phosphate ions results in the formation of calcium phosphate deposits oil the metal surface. In this Study, the deposits formed under various treatment conditions (pHs, concentrations and ratios of calcium/phosphate ions, and so on) were characterized by scanning electron spectroscopy (SEM). and X-ray diffractometry. The exclusive formation of hydroxyapatite, HAP, was observed under comparatively narrow conditions (pH 3-4, [Ca+]/[PO43-] = 25 mM/15 mM), which is clearly different from the reported conditions for the deposition of HAP on titanium substrates. HAP was deposited in the form of a layer. comprised of morphologically amorphous HAP flakes that were less than 20 nm thick. SEM and FTIR analyses of the deposit at different stages of H2O2-electrolysis revealed that a few dozen nanometer-sized spheres of amorphous calcium phosphate were formed in the first step and then fused with each other to form ribbon-like flakes of HAP or broken glass-like brushite, depending on the pH. The pH for HAP formation oil a stainless steel surface was markedly lower than that used for titanium, and the observed process by which amorphous calcium phosphate is converted to HAP was markedly different from that for the electrochemical deposition (electrolysis of water) of HAP on a titanium substrate. (C) 2009 Elsevier B.V. All rights reserved.

    DOI: 10.1016/j.colsurfa.2009.09.007

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  • Temperature Scanning FTIR Analysis of Secondary Structures of Proteins Embedded in Amorphous Sugar Matrix 査読

    Koreyoshi Imamura, Ken-Ichi Ohyama, Toru Yokoyama, Yoshinobu Maruyama, Nakanishi Kazuhiro

    JOURNAL OF PHARMACEUTICAL SCIENCES   98 ( 9 )   3088 - 3098   2009年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:JOHN WILEY & SONS INC  

    Heat-induced changes in secondary structures of five proteins (bovine serum albumin, BSA; human serum albumin, HSA; myoglobin; ribonuclease A, RNase A; and, beta-lactoglobulin, beta-Lg) in an amorphous sugar matrix were analyzed by temperature-scanning Fourier transform infrared spectroscopy to elucidate the mechanism of heat-induced conformational change of solid-phase proteins. Three sugars, trehalose, maltose, and dextran (MW 6000), were used. Loss of alpha-helices due to increasing temperature was observed for BSA, HSA, and myoglobin, which are rich in a-helices. RNase A showed a marked decrease in predominant secondary structural components (beta-sheet) with increasing temperature. However, no noticeable changes in the content of secondary structures, except for a slight loss of alpha-helices, were observed for beta-Lg, which is also beta-sheet-rich. These heat-induced conformational changes were significant at temperatures above the glass transition temperature. The heat-induced conformational change in BSA dried with sugar appeared time-independent and was clearly different from that due to dehydration and from the thermal conformational change for a solution of BSA. In particular, differences in secondary structural components that increased due to loss of alpha-helices were noted. (c) 2008 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 98:3088-3098, 2009

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  • Purification, Characterization, Molecular Cloning, and Expression of a New Aminoacylase from Streptomyces mobaraensis That Can Hydrolyze N-(Middle/Long)-chain-fatty-acyl-L-amino Acids as Well as N-Short-chain-acyl-L-amino acids 査読

    Mayuko Koreishi, Yasuyuki Nakatani, Manami Ooi, Hiroyuki Imanaka, Koreyoshi Imamura, Kazuhiro Nakanishi

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY   73 ( 9 )   1940 - 1947   2009年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:TAYLOR & FRANCIS LTD  

    We report here on the purification, characterization, molecular cloning, and expression of a new aminoacylase, initially isolated from the supernatant of Streptomyces mobaraensis (Sm-AA). Purified wild-type Sm-AA was found to be a monomeric protein with a molecular mass of 55kDa. The cloned gene of Sm-AA contained an ORF of 1,383 bp, encoding a polypeptide of 460 amino acids. A BLAST search revealed that Sm-AA belongs to the peptidase M20 family, with identities to a hypothetical protein from Streptomyces pristinaespiralis, a putative peptidase from Streptomyces averinitilis, peptidase M20 from Frankia sp., succinyl-diaminopimelate desuccinylase from Hemophilus influenzae, and aminoacylase-1 from porcine kidney at 89, 88, 67, 29, and 25% respectively. The Sm-AA gene was subcloned into an expression vector, pSH19, and was expressed in Streptomyces lividans TK24. The amount of the recombinant Sm-AA expressed in the S. lividans cells was approximately 42-fold higher than that of Sm-AA found in the supernatant of S. mobaraensis. Sm-AA showed high hydrolytic activity towards various N-acetyl-L-amino acids and N-(middle/long)-chain-fatty-acyl-L-amino acids, with a preference for the acyl derivatives of L-Met, L-Ala, L-Cys, etc. with an optimum pH and temperature for reaction of about 7.5 and 50 degrees C (at pH 7.5).

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  • Efficient N epsilon-lauroyl-L-lysine production by recombinant epsilon-lysine acylase from Streptomyces mobaraensis 査読

    Mayuko Koreishi, Ryoko Kawasaki, Hiroyuki Imanaka, Koreyoshi Imamura, Yasuaki Takakura, Kazuhiro Nakanishi

    JOURNAL OF BIOTECHNOLOGY   141 ( 3-4 )   160 - 165   2009年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE BV  

    epsilon-Lysine acylase from Streptomyces mobaraensis (Sm-ELA), which specifically catalyzes hydrolysis of the epsilon-amide bond in various N epsilon-acyl-L-lysines, was cloned and sequenced. The Sm-ELA gene consists of a 1617-bp open reading frame that encodes a 538-amino acid protein with a molecular mass of 55,816 Da. An NCBI protein-protein BLAST search revealed that the enzyme belongs to the YtcJ-like metal-dependent amidohydrolase family, which is further characterized as the metallo-dependent hydrolase superfamily. The Sm-ELA gene was ligated into a pUC702 vector for expression in Streptomyces lividans TK24. Expression of recombinant Sm-ELA in S. lividans was approximately 300-fold higher than that in wild-type S. mobaraensis. The recombinant Sm-ELAs from the cell-free extract and Culture supernatant were purified to homogeneity. The specific activities of the purified Sm-ELAs were 2500-2800 U/mg, which were similar to that obtained for the wild-type Sm-ELA. Using the cell-free extract of the recombinant S. lividans cells, N epsilon-lauroyl-L-lysine was synthesized from 500 mM L-lysine hydrochloride and 50, 100, or 250 mM lauric acid in an aqueous buffer Solution at 37 degrees C. The yields were close to 100% after 6 and 9 1) of reaction for 50 and 100 mM lauric acid, respectively. and 90% after 24 h for 250 mM lauric acid. (C) 2009 Elsevier B.V. All rights reserved.

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  • PS-tag連結タンパク質を利用したバイオ分子間相互作用解析

    今中 洋行, 上崎 英範, 國方 俊暢, 今村 維克, 中西 一弘

    化学工学会 研究発表講演要旨集   2009   934 - 934   2009年

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    記述言語:日本語   出版者・発行元:公益社団法人 化学工学会  

    DOI: 10.11491/scej.2009f.0.934.0

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  • 糖類アモルファス材料の水分収着特性とタンパク質安定化作用に及ぼす高圧処理の影響

    今村 維克, 野村 真世, 木村 佳文, 伊久 珠代, 絹畠 光倫, 今中 洋行, 中西 一弘

    化学工学会 研究発表講演要旨集   2009   779 - 779   2009年

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    記述言語:日本語   出版者・発行元:公益社団法人 化学工学会  

    DOI: 10.11491/scej.2009f.0.779.0

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  • クッションタンパク質を利用したバイオ分子固定化技術の開発と応用

    麻那古 秀友, 山隅 大輔, 大塚 隆尚, 今中 洋行, 今村 維克, 中西 一弘

    化学工学会 研究発表講演要旨集   2009   1060 - 1060   2009年

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    記述言語:日本語   出版者・発行元:公益社団法人 化学工学会  

    DOI: 10.11491/scej.2009f.0.1060.0

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  • オートトランスポーターを用いたバイオ分子表層提示系

    今中 洋行, 植田 久子, 山下 麻衣, 今村 維克, 中西 一弘

    化学工学会 研究発表講演要旨集   2009   606 - 606   2009年

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    記述言語:日本語   出版者・発行元:公益社団法人 化学工学会  

    DOI: 10.11491/scej.2009.0.606.0

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  • 糖類アモルファスマトリクスにおける糖-高分子間相互作用の形成特性

    今村 維克, 横山 徹, 大山 健一, 野村 真世, 今中 洋行, 中西 一弘

    化学工学会 研究発表講演要旨集   2009   608 - 608   2009年

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    記述言語:日本語   出版者・発行元:公益社団法人 化学工学会  

    DOI: 10.11491/scej.2009.0.608.0

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  • Recent advances in controlled immobilization of proteins onto the surface of the solid substrate and its possible application to proteomics 査読

    Kazuhiro Nakanishi, Takaharu Sakiyama, Yoichi Kumada, Koreyoshi Imamura, Hiroyuki Imanaka

    Current Proteomics   5 ( 3 )   161 - 175   2008年10月

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    記述言語:英語  

    Proteome analysis plays a key role in the elucidation of the functions and applications for numerous proteins. For proteome analyses, various microplate- and microarray-based techniques have been developed by a number of researchers. Their intent was to immobilize proteins on the surface of a solid substrate in a site-directed manner while retaining structure and native biological function. In this review, we focus on recent advances in immobilization methodology for proteins/enzymes on a surface, including those using the affinity peptides screened by random peptide library systems. We also discuss applications of the affinity peptide-mediated immobilization method in fields related to proteome analysis, particularly our recent work concerning immunoassay and protein-protein interaction analysis. ©2008 Bentham Science Publishers Ltd.

    DOI: 10.2174/157016408785909622

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  • Adsorption Characteristics of Various Proteins to a Titanium Surface 査読

    Koreyoshi Imamura, Miyuki Shimomura, Shungo Nagai, Megumi Akamatsu, Kazuhiro Nakanishi

    JOURNAL OF BIOSCIENCE AND BIOENGINEERING   106 ( 3 )   273 - 278   2008年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:SOC BIOSCIENCE BIOENGINEERING JAPAN  

    Adsorption characteristics of 18 proteins, with different sizes and isoelectric points, to a titanium oxide surface were studied. The adsorption isotherms were categorized based on protein type and pH: type 1, irreversible adsorption; type 2, Langmuir-type reversible adsorption; and type 3, reversible and irreversible adsorption. Most of the proteins tested were irreversibly adsorbed in the pH range of 3-8, whereas most adsorbed reversibly at pH 8.5-9.4. Protamine, with a pI value of 12, adsorbed reversibly in the pH range of 3-9. pH values that gave maximal sums of irreversibly and reversibly adsorbed proteins were in the pH range of 3-8 and tended to increase slightly with the pI value of the corresponding protein. pH values that gave maximal quantities of irreversibly adsorbed protein ranged between 4-6 and were nearly independent of pI.

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  • True density analysis of a freeze-dried amorphous sugar matrix 査読

    Koreyoshi Imamura, Yoshinobu Maruyama, Kazuhiro Tanaka, Tohru Yokoyama, Hiroyuki Imanaka, Kazuhiro Nakanishi

    JOURNAL OF PHARMACEUTICAL SCIENCES   97 ( 7 )   2789 - 2797   2008年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:JOHN WILEY & SONS INC  

    True density of an amorphous matrix represents the state of molecular packing in the matrix, which is closely related to the physical/chemical properties of the material. Dry gas pycnometry is one possible technique for measuring the true density of an amorphous sugar matrix prepared by freeze-drying. We herein report on the influence of conditions used for pycnometry on the measured density value and propose a protocol for obtaining the true density. The technique is sufficiently accurate to permit values for matrices comprised of different types of sugar to be compared. Using the protocol, the true densities of several amorphous sugar samples containing different types of sugar, freeze-drying conditions (temperature and sugar concentration at the time of freezing of an aqueous sugar solution), pretreatment (compaction and grind) were determined and the results were compared. A model for simulating an amorphous matrix of sugar (trehalose) was constructed using molecular dynamics/mechanics calculations, and the true density of the simulated sugar matrix was found to agree with the value experimentally determined using the proposed protocol. The relationship among the true density, the states of intermolecular interactions, and strain of sugar molecules in the matrix are discussed using the simulated amorphous sugar matrix. (C) 2007 Wiley-Liss, Inc.

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  • Characteristics of hydrogen bond formation between sugar and polymer in freeze-dried mixtures under different rehumidification conditions and its impact on the glass transition temperature 査読

    Koreyoshi Imamura, Yoko Asano, Yoshinobu Maruyama, Tohru Yokoyama, Mayo Nomura, Seiji Ogawa, Kazuhiro Nakanishi

    JOURNAL OF PHARMACEUTICAL SCIENCES   97 ( 3 )   1301 - 1312   2008年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:JOHN WILEY & SONS INC  

    The characteristics of hydrogen bond formation between trehalose and polyvinylpyrrolidone (PVP) in amorphous mixtures at different hydration states were quantitatively investigated. Amorphous trehalose-PVP mixtures were prepared by freeze-drying and equilibrated at different relative humidities (RH). Infrared (IR) spectra of the trehalose-PVP mixtures were obtained by Fourier transform IR spectroscopy,(FTIR) and the IRband corresponding to C=O groups of PVP was deconvolved into the component bands responsible for C=O groups that were free and restricted by hydrogen bonds, to estimate the degree of the trehalose-PVP interactions. The FTIR analysis indicated that approximately 80% of the C=O groups of PVP formed hydrogen bonds with trehalose in the presence of more than 3 g of trehalose per gramme of PVP, independent of the RH. IR analysis of the O-H stretching vibration of the sugar demonstrated that the presence of PVP lead to an increase in the free hydroxyl groups of trehalose that did not form hydrogen bonds at RH 0%. On the other hand, the water sorption behavior of the trehalose-PVP mixtures suggested that rehumidification diminished the effect of PVP on increasing the free OH groups. Thus a peculiar relationship may exist between T-g, RH and the composition of the mixture: The presence of PVP increased T-g at RHs 0 and above 23% but decreased T-g at 11%. (C) 2007 Wiley-Liss, Inc.

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  • Temperature scanning FTIR analysis of interactions between sugar and polymer additive in amorphous sugar-polymer mixtures 査読

    Koreyoshi Imamura, Ken-Ichi Ohyama, Toru Yokoyama, Yoshinobu Maruyama, Hiroyuki Imanaka, Kazuhiro Nakanishi

    JOURNAL OF PHARMACEUTICAL SCIENCES   97 ( 1 )   519 - 528   2008年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:JOHN WILEY & SONS INC  

    The impact of a polymer additive (polyvinylpyrrolidone, PVP) on hydrogen bonding in amorphous sugar matrices as well as on the glass transition temperature, T-g, were examined by temperature scanning Fourier transform infrared spectroscopy (TS-FTIR). An amorphous sugar matrix containing PVP was prepared by air-drying an aqueous solution of a sugar-PVP mixture. The hydrogen bonds in the sugar-PVP mixture (sugar-PVP and sugar-sugar hydrogen bonds) were analyzed from the IR peak positions corresponding to the stretching vibration of C=O groups of PVP and O-H groups of the sugar and the temperature dependence of the peak position of the O-H stretching vibration band. The addition of PVP to amorphous mono and disaccharides significantly lowered the extent of hydrogen bond formation while interactions between sugars and the PVP tended to prevent the disruption of hydrogen bonds due to increasing temperature, the magnitude of which was larger for larger oligomers. The T-g value for the amorphous sugar was increased by the addition of PVP in many cases. As the size of sugar molecule became larger, the relative magnitude of the increased T-g by PVP to the difference between the T-g values for sugar alone and PVP alone became larger and then reached a certain level; it was slight in the case of glucose. Collectively, these results demonstrate that the magnitude of the impact of PVP on an amorphous sugar matrix strongly vary and are dependent on the types of sugar. (C) 2007 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 97:519-528, 2008.

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  • Fourier self-deconvolution analysis of hydrogen bonding states of polyvinylpyrrolidone in an amorphous sugar matrix below and above the glass transition temperature 査読

    Koreyoshi Imamura, Ken-ichi Ohyama, Kazuko Tani, Toru Yokoyama, Yoshinobu Maruyama, Hiroyuki Imanaka, Kazuhiro Nakanishi

    SPECTROSCOPY LETTERS   41 ( 6 )   305 - 312   2008年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:TAYLOR & FRANCIS INC  

    In an amorphous mixture of sugar and polyvinylpyrrolidone (PVP), PVP carbonyl groups form hydrogen bonds with sugar hydroxyl groups, thereby improving the physical stability of the amorphous matrix against a glass-to-rubber transition. Herein, Fourier self-deconvolved IR bands due to the C=O stretching vibration of PVP in sugar-PVP mixtures were analyzed. The C=O groups in sugar-PVP mixtures generally had four vibrational states, corresponding with free and hydrogen-bonded C=O in three different modes. Changes in these vibrational states induced by increasing the temperature were compared among various sugar-PVP mixtures. Formation and thermal disruption characteristics of different modes of sugar-PVP hydrogen bondings are discussed.

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  • 過酸化水素の電気分解を利用した金属表面の洗浄技術であるH2O2-電気分解洗浄の洗浄特性に及ぼす諸因子の影響

    今村 維克

    日本食品工学会誌   9 ( 4 )   229 - 238   2008年

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    出版者・発行元:日本食品工学会  

    著者らは汚れの付着した金属表面上で集中的にOHラジカル (・OH) を発生させることを原理とする新たな洗浄手法を開発した.すなわち, 過酸化水素を含む水溶液に汚れの付着した金属表面を浸し, その状態で負電位を印加する.H2O2分子が金属表面で電子を受け取ることにより, きわめて酸化力が高い・OHが発生し, 汚れ物質を速やかに酸化分解し, 金属表面から除去することができる.すでに, タンパク質をモデル汚れ物質として, H2O2-電気分解洗浄における洗浄機構と有効性を検証している.本稿では, H2O2-電気分解洗浄における諸因子の影響の把握と洗浄効率のさらなる効率化を図るため, (1) 支持電解質の種類および濃度の影響, (2) 基板材質の影響, (3) 吸着物質 (タンパク質および脂質) の種類の影響, (4) 共存物質の影響について検討した.その結果, H2O2-電気分解洗浄においてアンモニウム化合物やリン酸イオンが存在するとモデル汚れ物質 (タンパク質) の除去速度が格段に向上することを明らかにした.また, 各種モデル汚れ物質や基板材質に対する有効性についても検証した.

    DOI: 10.11301/jsfe2000.9.229

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  • 高圧処理における糖類アモルファスマトリクスの物理特性変化

    今村 維克, 野村 真世, 横山 徹, 今中 洋行, 中西 一弘

    化学工学会 研究発表講演要旨集   2008   339 - 339   2008年

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    記述言語:日本語   出版者・発行元:公益社団法人 化学工学会  

    DOI: 10.11491/scej.2008.0.339.0

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  • 膜面液体培養法を用いた麹菌の培養特性及び遺伝子発現

    田中 創吉, 今中 洋行, 今村 維克, 中西 一弘

    化学工学会 研究発表講演要旨集   2008   378 - 378   2008年

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    記述言語:日本語   出版者・発行元:公益社団法人 化学工学会  

    DOI: 10.11491/scej.2008.0.378.0

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  • 親和性ペプチドを利用したポリスチレンおよびガラス表面への蛋白質固定化とその利用

    今中 洋行, 上崎 英範, 石村 遼太, 熊田 陽一, 今村 維克, 中西 一弘

    化学工学会 研究発表講演要旨集   2008   345 - 345   2008年

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    記述言語:日本語   出版者・発行元:公益社団法人 化学工学会  

    DOI: 10.11491/scej.2008.0.345.0

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  • クッションタンパク質を利用したバイオ分子固定化技術の開発

    山隅 大輔, 柳田 圭介, 今中 洋行, 今村 維克, 中西 一弘

    化学工学会 研究発表講演要旨集   2008   418 - 418   2008年

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    記述言語:日本語   出版者・発行元:公益社団法人 化学工学会  

    DOI: 10.11491/scej.2008.0.418.0

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  • 糖類アモルファスマトリクスの包括安定化作用および物理的諸特性に及ぼす高圧処理の影響

    今村 維克, 野村 真世, 田中 一宏, 木村 佳文, 今中 洋行, 中西 一弘

    化学工学会 研究発表講演要旨集   2008   175 - 175   2008年

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    記述言語:日本語   出版者・発行元:公益社団法人 化学工学会  

    DOI: 10.11491/scej.2008f.0.175.0

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  • ヒト由来転写因子とDNAとの相互作用解析システムの検討

    今中 洋行, 前川 真光, 今村 維克, 近藤 英作, 中西 一弘

    化学工学会 研究発表講演要旨集   2008   955 - 955   2008年

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    記述言語:日本語   出版者・発行元:公益社団法人 化学工学会  

    DOI: 10.11491/scej.2008f.0.955.0

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  • TPR domain of Ser/Thr phosphatase of Aspergillus oryzae shows no auto-inhibitory effect on the dephosphorylation activity 査読

    Bin Feng, Chun-Hui Zhao, Soukichi Tanaka, Hiroyuki Imanaka, Koreyoshi Imamura, Kazuhiro Nakanishi

    INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES   41 ( 3 )   281 - 285   2007年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE BV  

    A Ser/Thr phosphatase gene cloned from Aspergillus oryzae, aoppt, revealed that the tetratricopeptide repeat (TPR) and catalytic domains of the full-length AoPPT are located at the N- and C-terminal regions, respectively, similar to those of human Ser/Thr phosphatase 5 (PP5) and yeast Ppt1. Four different regions of AoPPT, namely, a full-length polypeptide, the catalytic domain, the catalytic domain plus C-terminal 15 amino-acid residues and the TPR domain were expressed in Escherichia coli and their roles in dephosphorylation activity were examined, using p-nitrophenyl phosphate as the substrate. The full-length AoPPT showed the highest dephosphorylation activity while the catalytic domain had the lowest activity. The activity of the catalytic domain was not inhibited by the presence of the TPR domain and arachidonic acid did not increase the activity of the full-length enzyme. These findings suggest that the integrity of the entire enzyme would be necessary for its full activity to be expressed. (c) 2007 Elsevier B.V. All rights reserved.

    DOI: 10.1016/j.ijbiomac.2007.03.005

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  • Enzymatic synthesis of beta-lactam antibiotics and N-fatty-acylated amino compounds by the acyl-transfer reaction catalyzed by penicillin V acylase from Streptomyces mobaraensis 査読

    Mayuko Koreishi, Kazuha Tani, Yuuichi Ise, Hiroyuki Imanaka, Koreyoshi Imamura, Kazuhiro Nakanishi

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY   71 ( 6 )   1582 - 1586   2007年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:TAYLOR & FRANCIS LTD  

    Penicillin V acylase from Streptomyces mobaraensis (Sm-PVA) showed high acyl-transfer activity in reactions using methyl esters of carboxylic acid (acyl donor) and amino compounds (nucleophile), to produce the corresponding amides. Moreover, Sm-PVA had broad substrate specificity, as indicated by the fact that it catalyzed the efficient synthesis of fl-lactam antibiotics, capsaicin derivatives, and N-fatty-acyl-amino acid/N-fatty-acyl-peptide derivatives.

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  • Cloning and characterization of penicillin V acylase from Streptomyces mobaraensis 査読

    Demin Zhang, Mayuko Koreishi, Hiroyuki Imanaka, Koreyoshi Imamura, Kazuhiro Nakanishi

    JOURNAL OF BIOTECHNOLOGY   128 ( 4 )   788 - 800   2007年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE BV  

    We report on the molecular cloning and characterization of penicillin V acylase (PVA) from an actinomycete, Streptomyces mobaraensis (Sm-PVA), which was originally isolated as an acylase that efficiently hydrolyzes the amide bond of various N-fatty-acyl-L-amino acids and N-fatty-acyl-peptides as well as capsaicin (8-methyl-N-vanillyl-6-nonenamide). In addition, the purified Sm-PVA hydrolyzed penicillin V with the highest activity (k(cat)) among the PVAs so far reported, penicillin G, and 2-nitro-5-phenoxyacetamide benzoic acid. The BLAST search revealed that the Sm-PVA precursor is composed of a polypeptide that is characteristic of enzymes belonging to the beta-lactam acylase family with four distinct segments; a signal sequence (43 amino acids), an a subunit (173 amino acids), a linker peptide (28 amino acids), and a beta subunit (570 amino acids). The mature, active Sm-PVA is a heterodimeric protein with alpha and beta subunits, in contrast to PVAs isolated from Bacillus sphaericus and B. subtilis, which have a homotetrameric structure. The amino acid sequence of Sm-PVA showed identities to PVA from S. lavendulae, N-acylhomoserine lactone-degrading acylase from Streptomyces sp., cyclic lipopeptide acylase from Streptomyces sp., and aculeacin A acylase from Actinoplanes utahensis with 68, 67, 67, and 41% identities, respectively. (c) 2007 Elsevier B.V. All rights reserved.

    DOI: 10.1016/j.jbiotec.2006.12.017

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  • Protein-protein interaction analysis using an affinity peptide tag and hydrophilic polystyrene plate 査読

    Yoichi Kumada, Chunhui Zhao, Ryota Ishimura, Hiroyuki Imanaka, Koreyoshi Imamura, Kazuhiro Nakanishi

    JOURNAL OF BIOTECHNOLOGY   128 ( 2 )   354 - 361   2007年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE BV  

    A sandwich ELISA method using peptide tags showing a specific affinity to a hydrophilic polystyrene surface (PS-tags), PS 19 composed of RAFIASRRIKRP and KPS19R10 of KRAFIASRRIRRP and a hydrophilic polystyrene (phi-PS) plate was used to analyze protein-protein interactions. An Escherichia coli cysteine synthase complex, in which serine acetyltransferase (SAT) interacts with O-acetylserine sulfhydrylase-A (OASS) was used as a model system. When the interaction was detected by the conventional sandwich ELISA method using a hydrophobic polystyrene (pho-PS) plate, for the exclusive use of ELISA, the signal intensity was barely detectable due to conformational change of the ligand protein, OASS in the adsorbed state. On the contrary, when OASS, genetically fused with PS19 (OASS-PS19) or chemically conjugated with KPS19R10 (OASS-KPS19R10), was immobilized on the phi-PS plate, a high signal intensity was detected. Furthermore, by applying the two-step sandwich ELISA, in which OASS-PS19 or OASS-KPS19R10 formed a complex with SAT in the blocking solution before immobilization on the phi-PS plate, the signal intensity was further increased with a much shorter operational time, because SAT in the blocking solution formed a complex with OASS-PS19 or OASS-KPS19R10 without any steric hindrance. (c) 2006 Elsevier B.V. All rights reserved.

    DOI: 10.1016/j.jbiotec.2006.09.018

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  • Development of a one-step ELISA method using an affinity peptide tag specific to a hydrophilic polystyrene surface 査読

    Yoichi Kumada, Shigeo Katoh, Hiroyuki Imartaka, Koreyoshi Imamura, Kazuhiro Nakanishi

    JOURNAL OF BIOTECHNOLOGY   127 ( 2 )   288 - 299   2007年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE BV  

    Glutathione S-transferase genetically fused with an affinity peptide tag, PS 19 (RAFIASRRIKRP) having a specific affinity for a hydrophilic polystyrene (PS) surface, was preferentially immobilized on a hydrophilic PS (phi-PS) plate without suffering from interference by coexisting protein molecules. Furthermore, rabbit IgG chemically conjugated with a peptide, KPS 19R10, in which (10)Lys in PS 19 was replaced with Arg and one Lys residue was added at the N-terminus as a coupling site for glutaraldehyde, showed a higher immobilization affinity to the phi-PS plate than that conjugated with the PS 19 peptide. On the basis of these findings, the use of a phi-PS plate and peptide tag-linked ligand proteins permitted a one-step or two-step enzyme-linked immumosorbent assay (ELISA) to be achieved, resulting in a substantial reduction in operational time compared with the conventional ELISA method using a hydrophobic PS (pho-PS) plate, while maintaining a high sensitivity. Furthermore, the sensitivity was increased to a greater extent compared to the conventional ELISA meihod when the one-step ELISA was applied to the detection of bovine insulin in a sandwich mode, due to the reduced number of washing and incubation steps. The method proposed here would be a versatile method for use in various ELISA techniques such as sandwich and competitive ELISAs using an antigen, an antibody and streptavidin that are genetically fused or chemically conjugated with the PS-specific affinity peptide as the ligand protein. (c) 2006 Elsevier B.V. All rights reserved.

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  • Adsorption characteristics of oligopeptides composed of acidic and basic amino acids on titanium surface 査読

    Koreyoshi Imamura, Yuuki Kawasaki, Takeshi Nagayasu, Takaharu Sakiyama, Kazuhiro Nakanishi

    JOURNAL OF BIOSCIENCE AND BIOENGINEERING   103 ( 1 )   7 - 12   2007年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:SOC BIOSCIENCE BIOENGINEERING JAPAN  

    The adsorption characteristics of octapeptides, containing different numbers of aspartic acid, lysine, and alanine residues (i.e., D(4)K(0)A(4), D(4)K(1)A(3), D(4)K(3)A(1), D(4)K(4)A(0), and D(0)K(4)A(4)) on the surface of titanium (Ti) particles were investigated in the pH range of 3.0-8.8 at 30 degrees C. The adsorption isotherms for octapeptides having four plural aspartic acid residues with or without lysine residues showed two distinct adsorption modes, i.e., irreversible and reversible modes, at pHs 3.0-6.5; at pH 7.0 or higher, the adsorption mode was reversible. Increasing the number of lysine residues at a fixed number of aspartic acid residues (i.e., 4) decreased the amount of peptides adsorbed in both modes. D(4)K(4)A(0) adsorbed irreversibly at pHs 3.0-6.5, due to the fact that negatively charged carboxyl groups directly interact with a positively charged Ti surface, whereas positively charged amino groups of lysine residues are directed in an opposite direction toward the solution side, as predicted by molecular mechanics/dynamics calculations.

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  • 過酸化水素-電気分解における金属表面でのハイドロキシアパタイト相の形成とその特性評価

    金本 和明, 今村 維克, 片岡 信秀, 押谷 潤, 今中 洋行, 中西 一弘

    化学工学会 研究発表講演要旨集   2007   126 - 126   2007年

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    記述言語:日本語   出版者・発行元:公益社団法人 化学工学会  

    DOI: 10.11491/scej.2007.0.126.0

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  • オートトランスポーターを利用した異種タンパク質の大腸菌細胞表層提示系の構築

    今中 洋行, 山下 麻衣, 盛永 鈴香, 今村 維克, 中西 一弘

    化学工学会 研究発表講演要旨集   2007   417 - 417   2007年

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    記述言語:日本語   出版者・発行元:公益社団法人 化学工学会  

    DOI: 10.11491/scej.2007.0.417.0

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  • PS-tagを用いたタンパク質間相互作用の検出

    熊田 陽一, Zhao Chunhui, 今中 洋行, 今村 維克, 中西 一弘

    化学工学会 研究発表講演要旨集   2007   427 - 427   2007年

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    記述言語:日本語   出版者・発行元:公益社団法人 化学工学会  

    DOI: 10.11491/scej.2007.0.427.0

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  • 糖-高分子複合アモルファスマトリクスにおける分子間相互作用と物理的安定性

    今村 維克, 浅野 容子, 大山 健一, 丸山 佳伸, 崎山 高明, 今中 洋行, 中西 一弘

    化学工学会 研究発表講演要旨集   2007   368 - 368   2007年

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    記述言語:日本語   出版者・発行元:公益社団法人 化学工学会  

    DOI: 10.11491/scej.2007.0.368.0

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  • オートトランスポーターを用いた大腸菌細胞表層提示系に及ぼすシグナルペプチドの影響

    今中 洋行, 辰本 渉, 今村 維克, 中西 一弘

    化学工学会 研究発表講演要旨集   2007   842 - 842   2007年

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    記述言語:日本語   出版者・発行元:公益社団法人 化学工学会  

    DOI: 10.11491/scej.2007f.0.842.0

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  • 大腸菌由来Cysteine synthaseの複合体形成特性

    今中 洋行, 森賀 雄大, 趙 春暉, 今村 維克, 中西 一弘

    化学工学会 研究発表講演要旨集   2007   841 - 841   2007年

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    記述言語:日本語   出版者・発行元:公益社団法人 化学工学会  

    DOI: 10.11491/scej.2007f.0.841.0

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  • 過酸化水素-電気分解により生成したハイドロキシアパタイトの特性評価および骨芽様細胞の増殖・分化特性

    今村 維克, 金本 和明, 今中 洋行, 中西 一弘

    化学工学会 研究発表講演要旨集   2007   990 - 990   2007年

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    記述言語:日本語   出版者・発行元:公益社団法人 化学工学会  

    DOI: 10.11491/scej.2007f.0.990.0

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  • H2O2-電気分解による金属表面および水溶液中の有機物質の分解・除去

    今村 維克, 黒田 俊樹, 小崎 正晴, 今中 洋行, 中西 一弘

    化学工学会 研究発表講演要旨集   2007   702 - 702   2007年

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    記述言語:日本語   出版者・発行元:公益社団法人 化学工学会  

    DOI: 10.11491/scej.2007f.0.702.0

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  • Identification of genes from Aspergillus oryzae that are preferentially expressed in membrane-surface liquid culture 査読

    Bin Feng, Masakazu Morita, Hiroyuki Imanaka, Koreyoshi Imamura, Kazuhlro Nakanishi

    JOURNAL OF BIOSCIENCE AND BIOENGINEERING   102 ( 5 )   470 - 473   2006年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:SOC BIOSCIENCE BIOENGINEERING JAPAN  

    We identified 22 genes from Aspergillus oryzae that are preferentially expressed in membrane-surface liquid culture (MSLC), among which Ser/Thr protein kinase (aopk1) and phosphatase (aoppt) genes were cloned. We also revealed that aopk1 encodes a protein with an N-terminal sequence 150 amino acid residues longer than that predicted from the registered sequence in GenBank.

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  • Effects of the supporting electrolyte on the kinetics of the removal of proteins adsorbed on a stainless steel surface by H2O2-electrolysis 査読

    Koreyoshi Imamura, Ippei Watanabe, Takaharu Sakiyama, Kazuhiro Nakanishi

    LANGMUIR   22 ( 16 )   7035 - 7040   2006年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER CHEMICAL SOC  

    The effects of different types of supporting electrolytes on the removal of beta-lactoglobulin (beta-Lg) after being adsorbed to a stainless steel surface by a H2O2-electrolysis treatment was investigated. In this process, hydroxyl radicals (center dot OH), generated by the electrolysis of hydrogen peroxide, decompose the substances adhering to the surface. The removal of the adsorbed protein from the stainless steel surface during the treatment was monitored in situ by ellipsometry. The apparent first-order removal rate constants, k(cl), for 17 types of supporting electrolytes were determined, as well as the current corresponding to the rate of generation of center dot OH. The kcl and generated current values for LiCl, NaCl, KCl, KNO3, K2SO4, CH3COOK, and K2CO3 were all similar. Ca2+ and Mg2+ strongly suppressed the removal of the adsorbed protein. The presence of ammonium compounds led to an increase in kcl and current values. In H2O2-electrolysis in the presence of potassium phosphate, the removal was extremely rapid, and an apparent increase in the thickness of the adsorbed layer was observed. The mechanisms responsible for the peculiar effects of calcium, magnesium, phosphate, and ammonium compounds were investigated by means of a Fourier transform infrared (FTIR) spectroscopic analysis, as well as by the characteristics of the removal under different treatment conditions.

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  • Temperature scanning FTIR analysis of hydrogen bonding states of various saccharides in amorphous matrixes below and above their glass transition temperatures 査読

    Koreyoshi Imamura, Keisuke Sakaura, Ken-ichi Ohyama, Atsushi Fukushima, Hiroyuki Imanaka, Takaharu Sakiyama, Kazuhiro Nakanishi

    JOURNAL OF PHYSICAL CHEMISTRY B   110 ( 31 )   15094 - 15099   2006年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER CHEMICAL SOC  

    Temperature scanning Fourier transform infrared, TS-FTIR, spectroscopy of various amorphous sugar matrixes was conducted to investigate the relationship between the glass transition temperature, T-g, of an amorphous sugar matrix and the nature of the hydrogen bonds in the matrix. An amorphous sugar matrix was prepared by air-drying an aqueous solution of sugar, and the degree of formation of hydrogen bonds in the matrix was evaluated at different temperatures using the peak positions of the IR band corresponding to the O-H stretching vibration at around 3400 cm(-1). The T-g value increased with increasing peak position of the O-H stretching vibration at T-g and were correlated reasonably well with the magnitude of the peak shift by the temperature increase (from 25 degrees C) to the T-g value. This demonstrates that the amorphous sugar matrix, in which the segments are fixed by fewer hydrogen bonds, has a higher thermal resistance. The glycosidic linkage largely contributes to the restriction of the segments, pyranose ring, rather than a hydrogen bond. As the degree of polymerization of pyranose rings increases, the degree of hydrogen bond formation needed to hold the matrix in a fixed position decreases. However, the magnitude of the restriction of pyranose rings by a glycosidic linkage changes depending on the type: the restrictions imposed by alpha-1,1 and -1,6 glycosidic linkages are the tightest and most flexible of all of the types of glycosidic linkages, respectively.

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  • Cloning, overexpression, purification, and characterization of O-acetylserine sulfhydrylase-B from Escherichia coli 査読

    C Zhao, Y Kumada, H Imanaka, K Imamura, K Nakanishi

    PROTEIN EXPRESSION AND PURIFICATION   47 ( 2 )   607 - 613   2006年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ACADEMIC PRESS INC ELSEVIER SCIENCE  

    O-Acetylserine sulfhydrylase-B (OASS-B, EC 2.5.1.47) is one of the two isozymes produced by Escherichia coli that catalyze the synthesis Of L-Cysteine from O-acetyl- L-serine and sulfide. The cysM gene encoding OASS-B was cloned and the enzyme was overexpressed in E coli using pUC19 with a lacUV5 promoter. The enzyme was purified to homogeneity, as evidenced by SDS-PAGE. Approximately 300 mg of purified OASS-B was obtained from 1600 mL of culture broth with a purification yield of 60% or higher. The purified OASS-13 was characterized and its properties compared with OASS-A. OASS-13 did not form a complex with E. coli serine acetyltransferase (SAT, EC 2.3.1.30) and showed a wide range of substrate specificity in nonproteinaceous amino acid synthesis. (c) 2006 Elsevier Inc. All rights reserved.

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  • Protease susceptibility of beta-lactoglobulin adsorbed on stainless steel surface as evidence of contribution of its specific segment to adsorption 査読

    Takaharu Sakiyama, Atsuko Aya, Mari Embutsu, Koreyoshi Imamura, Kazuhiro Nakanishi

    JOURNAL OF BIOSCIENCE AND BIOENGINEERING   101 ( 5 )   434 - 439   2006年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:SOC BIOSCIENCE BIOENGINEERING JAPAN  

    beta-Lactoglobulin (beta-Lg) is a major constituent of fouling deposits in the dairy industry. To determine the interaction between beta-Lg and stainless steel surfaces, beta-Lg irreversibly adsorbed on stainless steel particles was subjected to lysyl endopeptidase treatment and the course of fragmentation was compared with that observed for beta-Lg in solution. The results showed a distinct difference between the courses of fragmentation: a fragment (residues 102-135) was liberated readily from beta-Lg in solution but scarcely from beta-Lg irreversibly adsorbed on stainless steel particles. This result strongly suggests that residues 102-135 include a segment primarily responsible for the interaction of beta-Lg with stainless steel surfaces. This supports our previous results [Sakiyama et al., J. Biosci. Bioeng., 88, 536-541 (1999)] that showed that residues 125-135 of beta-Lg have a strong affinity toward stainless steel surfaces and probably a major contribution to the adsorption of beta-Lg.

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  • Screening and characterization of affinity peptide tags specific to polystyrene supports for the orientated immobilization of proteins 査読

    Y Kumada, Y Tokunaga, H Imanaka, K Imamura, T Sakiyama, S Katoh, K Nakanishi

    BIOTECHNOLOGY PROGRESS   22 ( 2 )   401 - 405   2006年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER CHEMICAL SOC  

    Dodecapeptides that exhibit a high affinity specific to a polystyrene surface (PS-tags) were screened using an Escherichia coli random peptide display library system, and the compounds were used as a peptide tag for the site-specific immobilization of proteins. The various PS-tags obtained after 10 rounds of biopanning selection were mainly composed of basic and aliphatic amino acid residues, most of which were arranged in close proximity to one another. Mutant-type glutathione S-transferases (GSTs) fused with the selected PS-tags. PS19 (RAFIASRRIKRP) and PS23 (AGLRLKKAAIHR) at their C-terminus, GST-PS19 and GST-PS23, when adsorbed on the PS latex beads had a higher affinity than the wild-type GST, and the specific remaining activity of the immobilized mutant-type GSTs was approximately 10 times higher than that of the wild-type GST. The signal intensity detected for GST-PS19 and GST-PS23 adsorbed on hydrophilic and hydrophobic PS surfaces using an anti-peptide antibody specific for the N-terminus peptide of GST was much higher than that for the wild-type GST. These findings indicate that the mutant-type GSTs fused with the selected peptide tags, PS19 and PS23, could be site-specifically immobilized on the surface of polystyrene with their N-terminal regions directed toward the solution. Thus, the selected peptide tags would be useful for protein immobilization in the construction of enzyme-linked immunosorbent assay (ELISA) systems and protein-based biochips.

    DOI: 10.1021/bp050331l

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  • On the interaction site of serine acetyltransferase in the cysteine synthase complex from Escherichia coli 査読

    CH Zhao, Y Moriga, B Feng, Y Kumada, H Imanaka, K Imamura, K Nakanishi

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   341 ( 4 )   911 - 916   2006年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ACADEMIC PRESS INC ELSEVIER SCIENCE  

    Cysteine synthase from Escherichia coli is a bienzyme complex comprised of serine acetyltransferase (SAT) and O-acetylserine sulfhydrylase A. The site of interaction of a SAT molecule was investigated by gel chromatography and surface plasmon technique using various mutant-type SATs, to better understand the mechanism involved in complex formation. The C-terminus of SAT, Ile 273, along with Glu 268 and Asp 271, was found to be essential for complex formation. The effects of O-acetyl-L-serine and sulfide on the affinity for the complex formation were also studied using a surface plasmon technique. (c) 2006 Elsevier Inc. All rights reserved.

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  • A novel acylase from Streptomyces mobaraensis that efficiently catalyzes hydrolysis/synthesis of capsaicins as well as N-Acyl-L-amino acids and N-acyl-peptides 査読

    M Koreishi, DM Zhang, H Imanaka, K Imamura, S Adachi, R Matsuno, K Nakanishi

    JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY   54 ( 1 )   72 - 78   2006年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER CHEMICAL SOC  

    A novel enzyme that catalyzes efficient hydrolysis of capsaicin (8-methyl-N-vanillyl-6-nonenamide) was isolated from the culture broth of Streptomyces mobaraensis. The enzyme consisted of two dissimilar subunits with molecular masses of 61 and 19 kDa. The enzyme was activated and stabilized in the presence of Co2+. It showed a pH optimum of about 8 and was stable at temperatures of up to 55 degrees C for 1 h at pH 7.8. The specific activity of the enzyme for the hydrolysis of capsaicin was 10(2)-10(4) times higher than those for the enzymes reported to date. In an aqueous/n-hexane biphasic system, capsaicin analogues such as octanoyl, decanoyl, and lauroyl vanillylamides were synthesized from the corresponding fatty acids and vanillylamine at yields of 50% or greater. In addition, the enzyme catalyzed the deacylation of N-lauroyl-L-amino acids and N-lauroyl-L-dipeptides and the efficient synthesis of N alpha-lauroyl-L-lysine, N epsilon-lauroyl-L-lysine, and various N-lauroyl-peptides in aqueous solution in both the absence and the presence of glycerol.

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  • 各種糖分子からなるアモルファスマトリクスの物理的諸特性

    今村 維克, 丸山 佳伸, 横山 徹, 大山 健一, 今中 洋行, 中西 一弘

    化学工学会 研究発表講演要旨集   2006   413 - 413   2006年

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    記述言語:日本語   出版者・発行元:公益社団法人 化学工学会  

    DOI: 10.11491/scej.2006f.0.413.0

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  • ポリスチレン親和性ペプチドを利用したタンパク質間相互作用の分析

    熊田 陽一, 石村 遼太, 上崎 英範, 今中 洋行, 今村 維克, 中西 一弘

    化学工学会 研究発表講演要旨集   2006   996 - 996   2006年

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    記述言語:日本語   出版者・発行元:公益社団法人 化学工学会  

    DOI: 10.11491/scej.2006f.0.996.0

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  • 金属表面へのタンパク質付着配向制御

    柳田 圭介, 中泉 雅人, 今中 洋行, 今村 維克, 中西 一弘

    化学工学会 研究発表講演要旨集   2006   1002 - 1002   2006年

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    記述言語:日本語   出版者・発行元:公益社団法人 化学工学会  

    DOI: 10.11491/scej.2006f.0.1002.0

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  • 放線菌 Streptomyces mobaraensis 由来アシラーゼによる転移反応とその応用

    是石 真友子, 谷 和葉, 伊勢 雄一, 今中 洋行, 今村 維克, 中西 一弘

    化学工学会 研究発表講演要旨集   2006   949 - 949   2006年

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    記述言語:日本語   出版者・発行元:公益社団法人 化学工学会  

    DOI: 10.11491/scej.2006f.0.949.0

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  • 高度な物理的安定性とタンパク質安定化作用を兼ね備えた糖類アモルファスマトリクスの創製

    横山 徹, 丸山 佳伸, 大山 健一, 今中 洋行, 今村 維克, 中西 一弘

    化学工学会 研究発表講演要旨集   2006   995 - 995   2006年

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    記述言語:日本語   出版者・発行元:公益社団法人 化学工学会  

    DOI: 10.11491/scej.2006f.0.995.0

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  • Purification and characterization of a novel aminoacylase from Streptomyces mobaraensis 査読

    M Koreishi, F Asayama, H Imanaka, K Imamura, M Kadota, T Tsuno, K Nakanishi

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY   69 ( 10 )   1914 - 1922   2005年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:TAYLOR & FRANCIS LTD  

    A novel aminoacylase was purified to homogeneity from culture broth of Streptomyces mobaraensis, as evidenced by SDS-polyacrylamide gel electrophoresis (PAGE). The enzyme was a monomer with an approximate molecular mass of 100 kDa. The purified enzyme was inhibited by the presence of 1,10-phenanthroline and activated by the addition of Co2+. It was stable at temperatures of up to 60 degrees C for 1 h at pH 7.2. It showed broad substrate specificity to N-acetylated L-amino acids. It catalyzed the hydrolysis of the amide bonds of various N-acetylated L-amino acids, except for N epsilon-acetyl-L-lysine and N-acetyl-L-proline. Hydrolysis of N-acetyl-L-methionine and N-acetyl-L-histidine followed Michaelis-Menten kinetics with K-m values of 1.3 +/- 0.1 mm and 2.7 +/- 0.1 mm respectively. The enzyme also catalyzed the deacetylation of 7-aminocephalosporanic acid (7-ACA) and cephalosporin C. Moreover, feruloyl-amino acids and L-lysine derivatives of ferulic acid derivatives were synthesized in an aqueous buffer using the enzyme.

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  • A novel ε-lysine acylase from Streptomyces mobaraensis for synthesis of Nε-acyl-L-lysines 査読

    Mayuko Koreishi, Ryoko Kawasaki, Hiroyuki Imanaka, Koreyoshi Imamura, Kazuhiro Nakanishi

    Journal of the American Oil Chemists’ Society   82 ( 9 )   631 - 637   2005年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1007/s11746-005-1121-2

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  • Adsorption characteristics of various organic substances on the surfaces of tantalum, titanium, and zirconium 査読

    T Nagayasu, K Imamura, K Nakanishi

    JOURNAL OF COLLOID AND INTERFACE SCIENCE   286 ( 2 )   462 - 470   2005年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ACADEMIC PRESS INC ELSEVIER SCIENCE  

    Adsorption characteristics of carboxylic acids, amines, an octapeptide composed of four L-alanine and four L-aspartic acid residues (Peptide-A(4)D(4)), and beta-lactoglobulin (beta-Lg) on tantalum (Ta), titanium (Ti), and zirconium (Zr) particles were examined at 30 degrees C and in some case, were compared with their adsorption onto SUS316L stainless steel particles (S6L). The adsorption isotherms on the Ta, Ti, and Zr particles could usually be expressed either by a Langmuir-type equation for reversible adsorption or by a modified Langmuir-type adsorption equation including terms for both reversible and irreversible adsorption. The adsorption equilibrium of benzoic acid, benzylamine, and m-xylylenediamine on all the metal surfaces followed a Langmuir-type equation, while those of phthalic acid, mellitic acid, and Peptide-A(4)D(4) could be fitted to the modified Langmuir-type adsorption equation. The adsorption characteristics of different adsorbates on the different surfaces were discussed particularly with reference to the pH dependencies of the q(irrev), q(rev), and K values and the electrostatic properties of the oxidized surface of the metal particles. Fourier transform infrared spectroscopic analyses using a reflection/absorption technique (RA-IR) indicated that phthalic acid and mellitic acid are adsorbed in similar adsorption states irrespective of the type of metal. beta-Lg was adsorbed onto the surfaces principally in an irreversible manner. The desorption behavior of beta-Lg from Ta, Ti, and S6L surfaces was examined, in order to evaluate the extent of interaction between beta-Lg and the metals. (c) 2005 Elsevier Inc. All rights reserved.

    DOI: 10.1016/j.jcis.2005.01.023

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  • Enzymatic Synthesis of Non-proteinaceous Amino Acids Catalyzed by Cysteine Synthase Coupled with Acetyl-CoA Regeneration

    Zhao Chunhui, Imanaka Hiroyuki, Imamura Koreyoshi, Nakanishi Kazuhiro

    化学工学会 研究発表講演要旨集   2005   271 - 271   2005年

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    記述言語:英語   出版者・発行元:公益社団法人 化学工学会  

    DOI: 10.11491/scej.2005.0.271.0

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  • 金属表面に吸着した各種タンパク質のH2O2-電気分解処理における脱離特性と脱離促進因子

    今村 維克, 尾下 学, 岩井 真澄, 金本 知明, 今中 洋行, 中西 一弘

    化学工学会 研究発表講演要旨集   2005   294 - 294   2005年

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    記述言語:日本語   出版者・発行元:公益社団法人 化学工学会  

    DOI: 10.11491/scej.2005.0.294.0

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  • 放線菌 Streptomyces mobaraensis 由来新規アシラーゼの精製・特性解析および合成反応

    是石 真友子, 川崎 涼子, 今中 洋行, 今村 維克, 中西 一弘, 丹尾 式希

    化学工学会 研究発表講演要旨集   2005   270 - 270   2005年

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    記述言語:日本語   出版者・発行元:公益社団法人 化学工学会  

    DOI: 10.11491/scej.2005.0.270.0

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  • 膜面液体培養法を用いた遺伝子組換え麹カビによるNuclease S1生産

    森田 正和, 安達 昇, 奥村 敦, 今中 洋行, 今村 維克, 中西 一弘

    化学工学会 研究発表講演要旨集   2005   242 - 242   2005年

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    記述言語:日本語   出版者・発行元:公益社団法人 化学工学会  

    DOI: 10.11491/scej.2005.0.242.0

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  • 温度走査フーリエ変換赤外分光分析による糖-高分子複合アモルファスマトリクスにおける分子間相互作用の解析

    大山 健一, 谷 加寿子, 今中 洋行, 今村 維克, 中西 一弘

    化学工学会 研究発表講演要旨集   2005   293 - 293   2005年

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    記述言語:日本語   出版者・発行元:公益社団法人 化学工学会  

    DOI: 10.11491/scej.2005.0.293.0

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  • 異なる金属表面への種々の有機物質の吸着挙動の比較

    長安 武司, 今中 洋行, 今村 維克, 中西 一弘

    化学工学会 研究発表講演要旨集   2005   292 - 292   2005年

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    記述言語:日本語   出版者・発行元:公益社団法人 化学工学会  

    DOI: 10.11491/scej.2005.0.292.0

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  • ポリスチレン親和性ペプチド融合タンパク質の吸着特性とその応用

    徳永 安秀, 熊田 陽一, 今石 大輔, 今中 洋行, 今村 維克, 崎山 高明, 中西 一弘

    化学工学会 研究発表講演要旨集   2005f   311 - 311   2005年

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    記述言語:日本語   出版者・発行元:公益社団法人 化学工学会  

    DOI: 10.11491/scej.2005f.0.311.0

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  • Characteristics of alpha-glucosidase production from recombinant Aspergillus oryzae by membrane-surface liquid culture in comparison with various cultivation methods 査読

    M Morita, H Shimamura, N Ishida, K Imamura, T Sakiyama, K Nakanishi

    JOURNAL OF BIOSCIENCE AND BIOENGINEERING   98 ( 3 )   200 - 206   2004年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:SOC BIOSCIENCE BIOENGINEERING JAPAN  

    alpha-Glucosidase was produced using recombinant Aspergillus oryzae by membrane-surface liquid culture (MSLC), a method previously developed by the authors and the results compared with other methods, including shaking flask culture (SFC), agar-plate culture (APC), culture on urethane sponge supports (USC), and liquid surface culture (LSC) to determine possible reasons for the advantageous features of MSLC. When yeast extract was used as a nitrogen source, the amount of enzyme produced by MSLC was 5 or more times higher than those for SFC and LSC, but similar to that using APC. Enzyme production in USC was slightly lower than in MSLC and APC. Cell growth was similar irrespective of the cultivation method used. When NaNO3, a typical inorganic nitrogen source was used, enzyme production in all the cultures was lower than that using yeast extract. However, even using NaNO3, the amount of the enzyme produced by MSLC was 8 to 20 times higher than those by SFC, APC, USC, and LSC. Although Cell growth using NaNO3 was similar to that for yeast extract in MSLC, it was markedly decreased in SFC, APC, and LSC. The reason for the difference in enzyme productivity for various cultivation methods using yeast extract and NaNO3 as a nitrogen source is discussed, on the basis of the experimental findings. The role of the oxygen transfer effect and gene expression levels in enzyme production were also examined.

    DOI: 10.1016/S1389-1723(04)00266-X

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  • Production of nonproteinaceous amino acids using recombinant Escherichia coli cells expressing cysteine synthase and related enzymes with or without the secretion of O-acetyl-L-serine 査読

    CH Zhao, K Ohno, K Sogoh, K Imamura, T Sakiyama, K Nakanishi

    JOURNAL OF BIOSCIENCE AND BIOENGINEERING   97 ( 5 )   322 - 328   2004年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:SOC BIOSCIENCE BIOENGINEERING JAPAN  

    beta-(Pyrazol-1-yl)-L-alanine (beta-PA), a model nonproteinaceous amino acid, was specifically synthesized by two methods using recombinant Escherichia coli cells that express cysteine synthase, comprising serine acetyltransferase (SAT) and O-acetylserine sulfhydrylase-A (OASS-A) and related enzymes from E. coli. In the first method (method A), recombinant cells that express wildtype SAT, OASS-A, acetate kinase (AK), and phosphotransacetylase (PTA) showed the highest beta-PA production. beta-PA was produced at 140 mM from 200 MM L-serine and 200 mM pyrazole under optimum conditions. Using the cells expressing SATDeltaC20 (truncated SAT), OASS-A, AK, and PTA, beta-PA was produced at a level of only 80 mM, whereas O-acetyl-serine (OAS) was found to be secreted into the broth. Under optimum conditions, OAS accumulated at levels of around 105 mM from 300 mM L-serine. Thus, in the second method (method B), the secreted OAS was used as the substrate for the syntheses of beta-PA and beta-(triazol-1-yl)-L-alanine (beta-TA). The OAS that accumulated in the broth was efficiently converted to beta-PA and beta-TA at levels of around 90 mM from 105 mM OAS using free OASS-A. In both methods A and B, the addition of glucose was essential for the efficient production of beta-PA and OAS, respectively.

    DOI: 10.1016/S1389-1723(04)70213-3

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  • Factors Affecting Performance of Cleaning Technique for Metal Surfaces Based on Electrolysis of Hydrogen Peroxide, H2O2-electrolysis 査読

    Koreyoshi Imamura

    nihon shokuhin kougakukaishi (japan journal of food engineering)   9 ( 4 )   229 - 238   2004年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    We have developed a novel technique for removing organic soilings from metal surfaces using hydroxy radicals (·OH). In the cleaning system, a metal surface fouled with organic soilings is made to contact with an aqueous solution containing hydrogen peroxide and supporting electrolyte. A slightly negative potential is then applied into the metal. The ·OHs, which are generated by the electrolysis of hydrogen peroxide (H2O2+e--ē·OH+OH”) on the metal surface, effectively attack and subsequently remove the adsorbed organic soils. In this study, the removal behavior of model organic soilings (protein) during the H2O2-electrolysis cleaning using different types and concentrations of supporting electrolyte was investigated. The presence of ammonium compounds and potassium phosphate led to marked increase in the removal rate. The influences of the types of metal substrate and adsorbed material on the removal behavior were also investigated, which suggested that the adsorption state of organic soiling on a metal surface strongly affect the removal characteristics. Furthermore, the removal rates in the presence of various organic coexisting materials were investigated. It was found that the removal rate constant was only slightly lowered in many cases. © 2008, Japan Society for Food Engineering. All rights reserved.

    DOI: 10.11301/jsfe2000.9.229

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  • Production of Non-Proteinaceous Amino Acids using Recombinant E. coli Cells

    Nakanishi Kazuhiro, Zhao Chunhui, Imamura Koreyoshi, Ohno Katsuhiro

    アジア・太平洋化学工学会議発表論文要旨集   2004   815 - 815   2004年

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    記述言語:英語   出版者・発行元:公益社団法人 化学工学会  

    β-(Pyrazol-1-yl)-L-alanine (β-pyrazole-L-Ala; β-PA), a model non-proteinaceous amino acid was synthesized using recombinant Escherichia coli cells that express cysteine synthase and enzymes that regenerate acetyl-CoA. In Method A, β-PA was synthesized from L-Ser and pyrazole using recombinant cells that express wild-type serine acetyltransferase (SAT), O-acetylserine sulfhydrylase-A (OASS-A), acetate kinase (AK), and phosphotransacetylase (PTA). Under the optimized condition, β-PA was produced at 140 mM from 200 mM L-Ser and 200 mM pyrazole. On the other hand, in Method B, O-acetyl-L-serine (OAS) was secreted into the broth, using the cells that express SATΔC20 (truncated SAT), OASS-A, AK, and PTA. Under optimized conditions OAS accumulated in the broth at levels of around 100 mM. The secreted OAS was efficiently converted to β-PA at levels of around 90 mM from 105 mM OAS using free OASS-A. In both Methods A and B, the addition of glucose was essential for the efficient production of β-PA and OAS, respectively.

    DOI: 10.11491/apcche.2004.0.815.0

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  • Removal Behavior of Protein Adsorbed on Metal Surface by H2O2-Electrolysis Treatment

    Imamura Koreyoshi, Watanabe Ippei, Aramoto Yumi, Sakiyama Takaharu, Nakanishi Kazuhiro

    アジア・太平洋化学工学会議発表論文要旨集   2004   353 - 353   2004年

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    記述言語:英語   出版者・発行元:公益社団法人 化学工学会  

    A novel method for generating hydroxyl radicals, ·OH, was developed, by applying slightly negative electric potentials (-0.2 ∼ -0.8 V vs Ag/AgCl) to the surface of a metal in contact with a hydrogen peroxide solution containing a supporting electrolyte. Namely, ·OH radicals are generated at the surface by the electrolysis of hydrogen peroxide according to the equation, H2O2 + e- -> ·OH + OH-. In this study, the kinetics of the removal of a model proteinaceous soiling (ß-lactoglobulin, ß-Lg) adsorbed to a stainless steel surface by the H2O2-electrolysis treatment were investigated. The rate of removal of the adsorbed ß-Lg from the stainless steel surface during the treatment was monitored in situ by ellipsometry, and the dependencies of the removal rate on H2O2 concentration, the electric potential applied to the surface, and the concentration and type of supporting electrolyte were examined.

    DOI: 10.11491/apcche.2004.0.353.0

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  • Effects of water activity and lipid addition on secondary structure of zein in powder systems 査読

    Y Mizutani, Y Matsumura, K Imamura, K Nakanishi, T Mori

    JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY   51 ( 1 )   229 - 235   2003年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER CHEMICAL SOC  

    The effects of water activity (A(w)) and lipid addition on the secondary structure of powdery zein were investigated using Fourier transform infrared spectroscopy. Two fatty acid esters, i.e., the linolenic and eicosapentaenoic acid ethyl esters (LAE and EPE), were mixed with the zein powder. The powders were stored in the "dry" state (with silica gel) and the "humid" state (A(w) = 0.9). The powdery zein without the lipids was shown to have a high content of the intermolecular hydrogen-bonded beta-sheet in the "dry" state, indicating the presence of protein aggregates. An increase in AW induced a decrease in this beta-sheet, concomitant with increases in the alpha-helix and beta-turn structures. The addition of LAE caused decreases in the alpha-helix and intermolecular hydrogen-bonded beta-sheet of zein when the powder was stored in the "humid" state, suggesting the strong interaction of LAE and zein molecules. However, LAE did not affect the secondary structure of zein in the "dry" state. The addition of EPE hardly influenced the secondary structure of zein, irrespective of A(w). These results are discussed in relation to the antioxidative activity of zein in the powder system, which had studied previously.

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  • ステンレス表面上の耐熱性アミノペプチダーゼの付着特性と活性発現

    長安 武司, 山越 憲吾, 吉岡 千里, 松本 訓智, 大西 宏輔, 今村 維克, 崎山 高明, 中西 一弘

    化学工学会 研究発表講演要旨集   2003   682 - 682   2003年

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    記述言語:日本語   出版者・発行元:公益社団法人 化学工学会  

    DOI: 10.11491/scej.2003f.0.682.0

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  • 温度走査FTIRによる各種糖類アモルファスマトリクスにおける相互作用の解析

    今村 維克, 坂浦 啓介, 福島 厚志, 大山 健一, 谷 加寿子, 崎山 高明, 中西 一弘

    化学工学会 研究発表講演要旨集   2003   688 - 688   2003年

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    記述言語:日本語   出版者・発行元:公益社団法人 化学工学会  

    DOI: 10.11491/scej.2003f.0.688.0

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  • Regulation of the Excretion of O-Acetyl-L-serine from Recombinant Escherichia coli Cells and Its Application to Synthesis of L-Cysteine and Non-protein Amino Acids

    ZHAO Chunhui, OHNO Katsuhiro, SOGOH Kohji, IMAMURA Koreyoshi, SAKIYAMA Takaharu, NAKANISHI Kazuhiro

    化学工学会 研究発表講演要旨集   2003   685 - 685   2003年

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    記述言語:日本語   出版者・発行元:公益社団法人 化学工学会  

    DOI: 10.11491/scej.2003f.0.685.0

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  • 膜面液体培養法(MSLC)と振盪フラスコ培養(SFC)を用いたα-glucosidase生産性の差についての考察

    森田 正和, 今村 維克, 崎山 高明, 中西 一弘

    化学工学会 研究発表講演要旨集   2003   681 - 681   2003年

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    記述言語:日本語   出版者・発行元:公益社団法人 化学工学会  

    DOI: 10.11491/scej.2003f.0.681.0

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  • 天然由来の高分子電解質を素材とした複合ゲルの調製とその膨潤特性

    崎山 高明, 今村 維克, 中西 一弘

    化学工学会 研究発表講演要旨集   2003   995 - 995   2003年

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    記述言語:日本語   出版者・発行元:公益社団法人 化学工学会  

    DOI: 10.11491/scej.2003f.0.995.0

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  • Isolation of a Novel Acylase from Streptomyces mobaraensis for Synthesis of Useful Amide Compounds

    Zhang Demin, Koreishi Mayuko, Imamura Koreyoshi, Sakiyama Takaharu, NAKANISHI Kazuhiro

    化学工学会 研究発表講演要旨集   2003   136 - 136   2003年

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    記述言語:日本語   出版者・発行元:公益社団法人 化学工学会  

    DOI: 10.11491/scej.2003f.0.136.0

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  • Enzymatic synthesis of N-acyl-L-amino acids in a glycerol-water system using acylase I from pig kidney 査読

    E Wada, M Handa, K Imamura, T Sakiyama, S Adachi, R Matsuno, K Nakanishi

    JOURNAL OF THE AMERICAN OIL CHEMISTS SOCIETY   79 ( 1 )   41 - 46   2002年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER OIL CHEMISTS SOC A O C S PRESS  

    N-Medium- and long-chain acyl-L-amino acids were enzymatically synthesized from the corresponding L-amino acids and fatty acids using a reverse hydrolysis. Enzymes that are suitable for the synthetic reaction of N-acyl-L-amino acids were screened on the basis of hydrolytic activity toward N-lauroyl-L-glutamic acid as an indicator. Acylase I from pig kidney (EC 3.5.1.14) showed the highest N-acyl-L-amino acid hydrolytic activity among 57 commercially available enzymes tested. Acylase I effectively catalyzed the synthesis of N-lauroyl-L-amino acids except for N-lauroyl-L-proline and N-lauroyl-L-tyrosine in a glycerol-water system. Under the optimized reaction conditions, N-lauroyl-L-arginine and N-lauroyl-L-glutamic acid were obtained in conversions of 82 and 44%, respectively. The equilibrium constants calculated from the conversion obtained were 5.6, 15.4, 18.0, and 39.4 for the syntheses of N-lauroyl-L-glutamic acid, N.-lauroyl-L-lysine, N-lauroyl-L-glutamine, and N-lauroyl-L-methionine, respectively. N-Acyl-L-arginines with myristic acid and palmitic acid as the fatty acid were also synthesized using acylase I.

    DOI: 10.1007/s11746-002-0432-7

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  • Enzymatic modification of beta-lactoglobulin with N-fatty-acyl-dipeptide by transglutaminase from Streptomyces mobaraense 査読

    E Wada, H Masuda, K Imamura, T Sakiyama, S Adachi, R Matsuno, K Nakanishi

    BIOTECHNOLOGY LETTERS   23 ( 17 )   1367 - 1372   2001年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:SPRINGER  

    beta -Lactoglobulin was enzymatically acylated with N-lauroyl-L-glutaminyl-glycine and N-lauroyl-L-glutamyl-L-lysine using transglutaminase from Streptomyces mobaraense. The modification of the protein with N-fatty-acyl-dipeptide was confirmed by SDS-PAGE, gel chromatography, HPLC, amino acid analysis, and TOF-MS. The degrees of the protein modification with N-lauroyl-L-glutaminyl-glycine and N-lauroyl-L-glutamyl-L-lysine were estimated to be 2-4 and 1.5 residues per molecule, respectively.

    DOI: 10.1023/A:1011699921628

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  • Role of sucrose-LDH hydrogen bond for thermal stabilizing effect of sucrose on freeze-dried LDH 査読

    T Suzuki, K Imamura, H Fujimoto, M Okazaki

    DRYING TECHNOLOGY   17 ( 7-8 )   1429 - 1439   1999年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:TAYLOR & FRANCIS INC  

    Relationship between thermal stabilizing effect of sugar on freeze-dried protein and sugar-protein hydrogen bond is studied. Sucrose and lactate dehydrogenase (LDH) are used as models for sugar and protein. Samples of freeze-dried LDH involved in sucrose of different crystallinity were prepared, and measurement of X-ray diffractometry (XRD) and Fourier-Transform Infrared (FT-IR) spectroscopy was done. It is found that when sucrose is amorphous, the degree of hydrogen bond formation is high and LDH is stabilized; when sucrose is crystalline, hydrogen bond is less formed and LDH is inactivated. These results indicate that the stabilizing effect of sugar is closely related to sugar-protein hydrogen bond. It is also found that there is an optimum sucrose content for the thermal stabilizing effect. This is because amorphous structure of sucrose is stabilized and protected from crystallization by LDH. Thus we can deduce that sugars and proteins work together to keep the activities of proteins.

    DOI: 10.1080/07373939908917625

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  • Relation between thermal stabilizing effect of sucrose on LDH and sucrose-LDH hydrogen bond 査読

    T Suzuki, K Imamura, H Fujimoto, M Okazaki

    JOURNAL OF CHEMICAL ENGINEERING OF JAPAN   31 ( 4 )   565 - 570   1998年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:SOC CHEMICAL ENG JAPAN  

    The thermal stabilizing effect of sugar on freeze-dried protein is studied, in particular on the relationship between the thermal stabilizing effect and the degree of sugar-protein hydrogen bond formation. Sucrose and lactate dehydrogenase (LDH) are used as models for sugar and protein. Several freeze-dried samples of LDH involved in sucrose were prepared; they differed in the degree of crystallinity of sucrose, By X-ray diffractometry (XRD) and Fourier transform infrared (FT-IR) spectroscopy, it is found that when sucrose is amorphous in the samples, the degree of hydrogen bond formation is high and LDH is stabilized remarkably. In contrast, when sucrose is crystalline, the degree of hydrogen bond formation is low and LDH is inactivated. These results indicate that the stabilizing effect of sugar is closely related to the sugar-protein hydrogen bond. The influence of sucrose content on the thermal stabilizing effect is also investigated. It is found that there is an optimum sucrose content for the thermal stabilizing effect. This is because, by the presence of LDH, the amorphous structure of sucrose is stabilized and protected from crystallization that causes loss of sucrose-LDH hydrogen bonds. Thus, we can deduce that sugars and proteins work together to maintain protein activities.

    DOI: 10.1252/jcej.31.565

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  • Thermal stabilization of freeze-dried enzymes by sugars 査読

    T Suzuki, K Imamura, K Yamamoto, T Satoh, M Okazaki

    JOURNAL OF CHEMICAL ENGINEERING OF JAPAN   30 ( 4 )   609 - 613   1997年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:SOC CHEMICAL ENG JAPAN  

    The thermal stabilization effect of sugars on freeze-dried proteins is studied, particularly the relation between stabilizing effect and the degree of sugar crystallinity. Three kinds of enzymes, alcohol dehydrogenase (ADH), lactate dehydrogenase (LDH), and malate dehydrogenase (MDH), were used as model proteins. Aqueous solutions of enzymes with sugars were freeze-dried and stored in dry air at 65 degrees C. The stabilities of freeze-dried enzymes improve remarkably by addition of trehalose or raffinose. By measurement of X-ray diffractometry, the sugars are found to form fully amorphous matrix in freeze-dried samples. Furthermore, sucrose stabilizes enzymes to a great extent when it is amorphous in samples, though it shows Little stabilizing effect when it is crystalline. These results indicate that the stabilizing effect of sugars closely relates to the amorphous matrix formed by the sugars.

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  • Stabilizing effect on freeze-dried proteins by amorphous matrices of sugars 査読

    T Suzuki, K Imamura, K Yamamoto, M Okazaki

    DRYING '96, VOL B   1261 - 1266   1996年

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    記述言語:英語   掲載種別:研究論文(国際会議プロシーディングス)   出版者・発行元:DRUKARNIA PAPAJ  

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  • A New Method for Measurement of Powder Characteristics Based on Reentrainment Phenomena 査読

    Hiroaki Masuda, Shuji Matsusaka, Koreyoshi Imamura

    KONA Powder and Particle Journal   12   133 - 143   1994年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    A new method has been developed to measure and evaluate the operational powder characteristics for fine particle processing based on the reentrainment phenomena. Experiments on particle reentrainment were carried out using an accelerated air flow for 21 different test powders. Both the reentrainment fluxes, which were measured by an electrostatic method, and the average air velocities were automatically sampled by a computer. The sampled data were processed to obtain the reentrainment profiles as a function of the average air velocity. The cumulative reentrainment efficiencies were also obtained and represented as a function of the wall shear stress. It was found that fine particles having tendencies to form large aggregates were reentrained mainly in the early stage of the measurement when the flow velocity was as low as 5 m·s−1. Also, the mass ratio of the reentrained large aggregates to the total particles was used to obtain information on the particle-particle (cohesion) or particle-wall interactions (adhesion). On the other hand, the cumulative reentrainment efficiency-curves must be applied to determine the operational conditions controlling the amount of adhered particles in various aerosol processes. © 1994, Hosokawa Powder Technology Foundation. All rights reserved.

    DOI: 10.14356/kona.1994021

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  • 飛散現象を利用した粉体特性の測定と評価

    増田 弘昭, 松坂 修二, 今村 維克

    粉体工学会誌   29 ( 12 )   897 - 905   1992年

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    記述言語:日本語   出版者・発行元:The Society of Powder Technology, Japan  

    A new method has been developed to measure and evaluate the operational powder characteristics for fine particle processing based on the reentrainment phenomena. Experiments on particle reentrainment were carried out by use of an accelerated air flow for 21 different test powders. Both the reentrainment fluxes, which were measured by an electrostatic method, and the average air velocities were automatically sampled in a computer. The sampled data were processed to give the reentrainment profiles as a function of the average air velocity. The cumulative reentrainment efficiencies were also obtained and represented by a function of the wall shear stress.<br>It was found that fine particles having tendencies to form large aggregates were reentrained mainly in the early stage of the measurement where the flow velocity was as low as 5m·s-1. Also, the mass ratio of the reentrained large aggregates to the total particles was well applied to obtain the information on the particle-particle or particle-wall interactions. On the other hand, the cumulative reentrainment efficiency-curves would be applied to determine the operational conditions controlling the amount of adhered particles in various aerosol processes.

    DOI: 10.4164/sptj.29.897

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MISC

  • パッキング相互作用に着目したタンパク質結晶化法

    小坂恵, 山田秀徳, 二見淳一郎, 多田宏子, 今村維克, 玉田太郎

    日本結晶学会年会講演要旨集   2019   2019年

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  • パッキングのコントロールにより,難結晶性蛋白質の結晶化を促進する結晶化法

    小坂恵, 山田秀徳, 二見淳一郎, 多田宏子, 今村維克, 玉田太郎

    日本細胞生物学会大会(Web)   71st   2019年

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  • 難結晶性蛋白質のパッキングをコントロールする結晶化法の開発と評価

    小坂恵, 山田秀徳, 二見淳一郎, 多田宏子, 今村維克, 玉田太郎

    日本結晶学会年会講演要旨集   2018   2018年

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  • タンパク質のパッキングをコントロールする疎水性残基導入

    小坂恵, 山田秀徳, 二見淳一郎, 多田宏子, 今村維克, 玉田太郎

    日本結晶学会年会講演要旨集   2017   107   2017年11月

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  • 疎水性残基導入が難結晶性タンパク質の結晶化に与える影響

    小坂恵, 山田秀徳, 二見淳一郎, 多田宏子, 今村維克, 玉田太郎

    日本生化学会大会(Web)   90th   ROMBUNNO.3P‐0100 (WEB ONLY)   2017年

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  • Sugar surfactantの応用 : 油滴包括およびタンパク質安定化

    今村 維克, 石田 尚之, 今中 洋行

    日本食品工学会誌 = Japan journal of food engineering   17 ( 3 )   A - 7-11   2016年9月

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    記述言語:日本語   出版者・発行元:日本食品工学会  

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  • 糖の有機溶媒に対する過溶解現象を利用した香気成分の糖類アモルファス固体中への界面活性物質free固体分散化技術の開発

    今村 維克

    食に関する助成研究調査報告書   ( 28 )   67 - 76   2015年

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    記述言語:日本語   出版者・発行元:すかいらーくフードサイエンス研究所  

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  • 2P-064 新規Pd 親和性ペプチドの探索およびタグ連結タンパク質の固定化特性解析(タンパク質工学,一般講演)

    的場 晴香, 石田 尚之, 今村 維克, 今中 洋行

    日本生物工学会大会講演要旨集   67   190 - 190   2015年

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    その他リンク: http://search.jamas.or.jp/link/ui/2016231956

  • 3P-195 5'末端周辺配列および培地組成が大腸菌発現系に及ぼす影響(バイオプロセス,一般講演)

    安保 紘高, 原 啓文, 梅津 光央, 石田 啓文, 今村 維克, 今中 洋行

    日本生物工学会大会講演要旨集   67   319 - 319   2015年

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    記述言語:日本語   出版者・発行元:日本生物工学会  

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    その他リンク: http://search.jamas.or.jp/link/ui/2016232284

  • 標的遺伝子の5’末端周辺配列が大腸菌発現系に及ぼす影響

    安保紘高, 原啓文, 石田尚之, 今村維克, 今中洋行

    日本農芸化学会大会講演要旨集(Web)   2015   2015年

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  • 2P-020 大腸菌発現系において培地および遺伝子配列が発現量および転写プロファイルに及ぼす影響(遺伝子工学,一般講演)

    安保 紘高, 三木 駿也, 原 啓文, 山副 敦司, 細山 哲, 土金 恵子, 今村 維克, 今中 洋行

    日本生物工学会大会講演要旨集   66   111 - 111   2014年

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    その他リンク: http://search.jamas.or.jp/link/ui/2016045901

  • Utilization of Hydrophilic Polystyrene Affinity Peptide for Functional Biomolecular Immobilization and Effective Site Specific Peptide Binder Screening

    H. Imanaka, R. Matsushita, T. Takimoto, T. Kunikata, K. Imamura, K. Nakanishi

    BIOPOLYMERS   100 ( 3 )   270 - 270   2013年5月

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    記述言語:英語   掲載種別:研究発表ペーパー・要旨(国際会議)   出版者・発行元:WILEY-BLACKWELL  

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  • 3P-180 新規金表面親和性ペプチドの同定及びタンパク質固定化への応用(生物化学工学,一般講演)

    重森 陽士郎, 石田 尚之, 今村 維克, 高橋 裕一郎, 今中 洋行

    日本生物工学会大会講演要旨集   65   233 - 233   2013年

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  • 2P-076 リガンド分子の固定化配向および密度がバイオ分子間相互作用に及ぼす影響(パンパク質工学,一般講演)

    松下 瑠奈, 石田 尚之, 今村 維克, 今中 洋行

    日本生物工学会大会講演要旨集   65   123 - 123   2013年

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    その他リンク: http://search.jamas.or.jp/link/ui/2014247513

  • 4Cp02 機能的バイオ分子固定化技術を利用したNFkB(p50)親和性ペプチドの探索(タンパク質工学/核酸工学,一般講演)

    瀧本 貴之, 宮原 徹也, 今村 維克, 今中 洋行, 近藤 英作, 中西 一弘

    日本生物工学会大会講演要旨集   64   195 - 195   2012年

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    その他リンク: http://search.jamas.or.jp/link/ui/2013125671

  • 4Cp03 転写因子FOXP3を標的とした部位特異的ペプチド薬剤スクリーニング技術の検討(タンパク質工学/核酸工学,一般講演)

    川崎 一起, 松本 亘平, 今中 洋行, 今村 維克, 近藤 英作

    日本生物工学会大会講演要旨集   64   195 - 195   2012年

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  • 11p04 大腸菌を宿主としたStreptomyces mobaraensis由来ε-lysine acylase大量発現系の検討(生物化学工学,一般講演)

    今中 洋行, 田口 友造, 内田 達也, 公文 一輝, 黒木 健太郎, 今村 維克, 中西 一弘

    日本生物工学会大会講演要旨集   63   58 - 58   2011年

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  • 2Ep03 機能的タンパク質固定化技術を利用したNFkB(p50)阻害性ペプチドの探索(タンパク質工学/核酸工学/ペプチド工学,一般講演)

    瀧本 貴之, 宮原 徹也, 今中 洋行, 今村 維克, 近藤 英作, 中西 一弘

    日本生物工学会大会講演要旨集   63   145 - 145   2011年

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  • Development of a highly efficient indigo dyeing method using indican with an immobilized β-glucosidase from Aspergillus niger

    Song Jingyuan, Imanaka Hiroyuki, Imamura Koreyoshi, KAJITANI Kouichi, NAKANISHI Kazuhiro

    Journal of bioscience and bioengineering   110 ( 3 )   281 - 287   2010年9月

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    記述言語:英語   出版者・発行元:公益社団法人日本生物工学会  

    A highly efficient method for dyeing textiles with indigo is described. In this method, the substrate, indican is first hydrolyzed at an acidic pH of 3 using an immobilized β-glucosidase to produce indoxyl, under which conditions indigo formation is substantially repressed. The textile sample is then dipped in the prepared indoxyl solution and the textile is finally exposed to ammonia vapor for a short time, resulting in rapid indigo dyeing. As an enzyme, we selected a β-glucosidase from Aspergillus niger, which shows a high hydrolytic activity towards indican and was thermally stable at temperatures up to 50-60℃, in an acidic pH region. The A. niger β-glucosidase, when immobilized on Chitopearl BCW-3001 by treatment with glutaraldehyde, showed an optimum reaction pH similar to that of the free enzyme with a slightly higher thermal stability. The kinetics for the hydrolysis of indican at pH 3, using the purified free and immobilized enzymes was found to follow Michaelis-Menten type kinetics with weak competitive inhibition by glucose. Using the immobilized enzyme, we successfully carried out repeated-batch and continuous hydrolyses of indican at pH 3 when nitrogen gas was continuously supplied to the substrate solution. Various types of model textiles were dyed using the proposed method although the color yield varied, depending on the type of textile used.

    DOI: 10.1016/j.jbiosc.2010.03.010

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  • 糖類アモルファスマトリクスの温度走査フーリエ変換赤外分光分析 : 相互作用状態の温度依存性

    今村 維克

    熱測定 : netsu = Calorimetry and thermal analysis   37 ( 4 )   154 - 161   2010年8月

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    記述言語:日本語   出版者・発行元:日本熱測定学会  

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  • Effects of the Cultivation Method on the Characteristics or Gene Expression Profiles of Aspergillus oryzae Using mCD or DPY Media

    Imanaka Hiroyuki, Tanaka Soukichi, Feng Bin, Imamura Koreyoshi, Nakanishi Kazuhiro

    Memoirs of the Faculty of Engineering, Okayama University   44   73 - 83   2010年1月

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    記述言語:英語   出版者・発行元:Faculty of Engineering, Okayama University  

    We used modified Czapek-Dox (mCD) or dextrin-peptone-yeast extract (DPY) media to cultivate a filamentous fungus, Aspergillus oryzae IAM 2706 by three different cultivation methods, i.e., shaking-flask culture (SFC), agar-plate culture (APC), and membrane-surface liquid culture (MSLC), to identify the differences in cultivation behaviors and gene transcriptional profiles. The fungi cultivated by APC or MSLC secreted a greater number of different proteins/enzymes in larger quantities compared with fungi cultivated by SFC, particularly when DPY medium was used. In particular, the amounts of protease secreted by fungi cultivated via MSLC or APC were much greater compared with SFC. When mCD medium was used, α-amylase activity was barely detectable in all cultures while the activity was detected in MSLC and APC in a quantity that was several times higher than that in SFC using DPY medium. SDS-PAGE analysis and N-terminal amino acid sequences confirmed 6 proteins in the culture supernatants when DPY medium was used. Among these proteins oryzin (an alkaline protease) and α-amylase were detected at much higher levels in APC and MSLC compared with SFC, which was consistent withthe measured activity of the secreted enzymes. However, when mCD medium was used, only oryzin was detected in significant amounts in MSLC and APC. Microarray analyses of the fungi cultivated by SFC, APC or MSLC using either mCD or DPY media indicated that the gene transcriptional profile of the MSLC sample was similar to that of the APC sample but different from that of the SFC sample. When mCD medium was used, most of the genes that were up-regulated 10-folds or greater in the MSLC sample relative to the SFC sample were unknown or predicted proteins. Transcription of the oryzin gene was only slightly up-regulated in the MSLC sample while transcription of the α-amylase gene was slightly down-regulated. On the other hand, when DPY medium was used, many known genes including the oryzin gene were up-regulated in the MSLC sample versus the SFC sample.

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  • 2P-1103 オートトランスポーターを用いた大腸菌細胞表層提示系におけるアミノ酸配列の影響(2bタンパク質工学,一般講演,酵素学,タンパク質工学および酵素工学,伝統の技と先端科学技術の融合)

    今中 洋行, 植田 久子, 今村 維克, 中西 一弘

    日本生物工学会大会講演要旨集   22   34 - 34   2010年

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  • 3P-1055 Multiformity of β-Galactosidase Produced from Bacillus circulans :

    SONG Jingyuan, IMANAKA Hiroyuki, IMAMURA Koreyoshi, MINODA Masashi, YAMAGUCHI Shoutaro, NAKANISHI Kazuhiro

    日本生物工学会大会講演要旨集   22   71 - 71   2010年

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  • 3P-1085 機能的ペプチド固定化法を利用したCysteine Synthase複合体形成における機能性残基の同定(2bタンパク質工学,一般講演,酵素学,タンパク質工学および酵素工学,伝統の技と先端科学技術の融合)

    清水 友樹, 今中 洋行, 今村 維克, 中西 一弘

    日本生物工学会大会講演要旨集   22   79 - 79   2010年

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  • MN-P23 Development of peptide immobilization method using PS-tagged cushion protein(Section X Micro/Nano Technology for Analysis and Cell Manipulation)

    Imanaka Hiroyuki, Yamazumi Daisuke, Kunikata Toshinobu, Imamura Koreyoshi, Nakanishi Kazuhiro

    Journal of bioscience and bioengineering   108 ( 1 )   S161   2009年11月

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  • 酵素糖化と発酵を併用した小麦フスマからの効率的エタノール生産

    是石 真友子, 今中 洋行, 今村 維克, 狩山 昌弘, 中西 一弘

    生物工学会誌 : seibutsu-kogaku kaishi   87 ( 5 )   216 - 223   2009年5月

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    記述言語:日本語   出版者・発行元:日本生物工学会  

    小麦フスマを原料として用い,市販酵素製剤による糖化反応と酵母によるエタノール発酵を併用した効率的エタノール生産方法の構築のために必要なバイオマスの前処理条件および糖化のための酵素の組み合わせなどの検討を行った.糖化反応条件としては,小麦フスマを140℃で保持時間0hの条件下で水熱処理を行い,反応温度45℃でMeiselaseおよびNovozyme^[○!R]188の2種類の酵素製剤を用いて糖化することが,グルコース生成に対して最も効果的であることがわかった.3.3%(w/v)の水熱処理小麦フスマから理論値の90%に相当するグルコースが生成した.しかし,33%(w/v)の水熱処理小麦フスマを用いた場合には,理論値の72%のグルコースしか生成しなかった.この理由として,反応生成物であるグルコースのβ-グルコシダーゼに対する阻害の影響が考えられた.しかし,酵素糖化を45℃で24時間行った後に,酵母を添加し30℃でエタノール発酵を行うと,糖化反応とグルコース消費が併行して起こるために阻害効果が緩和され,その結果,理論値の88%に相当する約5.2%(w/v)のエタノールが生成した.

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  • 2Ea05 オートトランスポーターを用いたペプチド細胞表層提示系の検討(タンパク質工学,一般講演)

    植田 久子, 山下 麻衣, 今中 洋行, 今村 維克, 中西 一弘

    日本生物工学会大会講演要旨集   21   59 - 59   2009年

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  • 2Ip05 クッションタンパク質を利用したペプチドータンパク質間相互作用検出系の検討(バイオセンシング・分析化学,一般講演)

    今中 洋行, 國方 俊暢, 山之内 麻衣, 今村 維克, 中西 一弘

    日本生物工学会大会講演要旨集   21   126 - 126   2009年

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  • 2Da01 放線菌Streptomyces mobaraensis由来新規アミノアシラーゼの発現系の構築および特性解析(酵素学・酵素工学,一般講演)

    多井 愛美, 中谷 泰之, 是石 真友子, 今中 洋行, 今村 維克, 中西 一弘

    日本生物工学会大会講演要旨集   21   44 - 44   2009年

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  • 3Ga07 ヒトFOXP3-DNA相互作用の迅速検出システムの構築(酵素学,酵素工学,タンパク質工学,糖鎖工学,一般講演)

    前川 真光, 今中 洋行, 今村 維克, 近藤 英作, 中西 一弘

    日本生物工学会大会講演要旨集   20   186 - 186   2008年

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    その他リンク: http://search.jamas.or.jp/link/ui/2009066377

  • 2Aa03 組換えモノグリセリドリパーゼの精製及び生化学的特性解析(プロセス工学,一般講演)

    寺川 秀嗣, 森 真基, 今中 洋行, 今村 維克, 中西 一弘

    日本生物工学会大会講演要旨集   20   59 - 59   2008年

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  • 3Da04 クッションタンパク質を用いたバイオ分子固定化技術(分析化学・物理化学,一般講演)

    今中 洋行, 山隅 大輔, 柳田 圭介, 今村 維克, 中西 一弘

    日本生物工学会大会講演要旨集   20   126 - 126   2008年

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  • 新規アシラーゼを用いた食品タンパク質のプロテアーゼ消化ペプチド断片への脂肪酸付加と機能性の調査

    中西 一弘, 今村 維克, 今中 洋行

    食に関する助成研究調査報告書   ( 21 )   109 - 115   2008年

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    記述言語:日本語   出版者・発行元:すかいら-くフ-ドサイエンス研究所  

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  • Enzymatic Synthesis of β-Lactam Antibiotics and N-Fatty-Acylated Amino Compounds by the Acyl-Transfer Reaction Catalyzed by Penicillin V Acylase from Streptomyces mobaraensis

    Koreishi Mayuko, Tani Kazuha, Ise Yuuichi, IMANAKA Hiroyuki, IMAMURA Koreyoshi, NAKANISHI Kazuhiro

    Bioscience, biotechnology, and biochemistry   71 ( 6 )   1582 - 1586   2007年6月

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    記述言語:英語   出版者・発行元:公益社団法人 日本農芸化学会  

    Penicillin V acylase from &lt;I&gt;Streptomyces mobaraensis&lt;/I&gt; (&lt;I&gt;Sm&lt;/I&gt;-PVA) showed high acyl-transfer activity in reactions using methyl esters of carboxylic acid (acyl donor) and amino compounds (nucleophile), to produce the corresponding amides. Moreover, &lt;I&gt;Sm&lt;/I&gt;-PVA had broad substrate specificity, as indicated by the fact that it catalyzed the efficient synthesis of &amp;beta;-lactam antibiotics, capsaicin derivatives, and &lt;I&gt;N&lt;/I&gt;-fatty-acyl-amino acid/&lt;I&gt;N&lt;/I&gt;-fatty-acyl-peptide derivatives.

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  • 2F14-4 膜面液体培養法と振盪フラスコ培養法を用いた麹菌のプロテアーゼ生産及び遺伝子発現解析(培養工学,センサー・計測工学・ロボット工学,プロセス工学,一般講演)

    田中 創吉, 馮 斌, 今中 洋行, 今村 維克, 中西 一弘

    日本生物工学会大会講演要旨集   19   141 - 141   2007年

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    その他リンク: http://search.jamas.or.jp/link/ui/2008290979

  • 1C11-1 放線菌Streptomyces mobaraensis由来長鎖および短鎖アミノアシラーゼの特性解析およびクローニング(酵素学・酵素工学・タンパク質工学,一般講演)

    中谷 泰之, 是石 真友子, 今中 洋行, 今村 維克, 中西 一弘

    日本生物工学会大会講演要旨集   19   79 - 79   2007年

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  • 1C09-2 オートトランスポーターを用いた細胞表層提示系の構築におけるシグナルペプチドの影響(酵素学・酵素工学・タンパク質工学,一般講演)

    辰本 渉, 今中 洋行, 今村 維克, 中西 一弘

    日本生物工学会大会講演要旨集   19   77 - 77   2007年

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  • 1C11-2 放線菌Streptomyces mobaraensis由来ペニシリン V アシラーゼのアシル基転移反応を利用した機能性化合物の酵素合成(酵素学・酵素工学・タンパク質工学,一般講演)

    是石 真友子, 伊勢 雄一, 谷 和葉, 今中 洋行, 今村 維克, 中西 一弘

    日本生物工学会大会講演要旨集   19   79 - 79   2007年

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  • ポリスチレン親和性ペプチド標識タンパク質を用いる迅速免疫測定

    中西一弘, 熊田陽一, 今中洋行, 今村維克

    日本食品工学会年次大会講演要旨集   8th   2007年

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  • "ガラス" によるタンパク質保存技術の可能性

    今村 維克

    冷凍 = Refrigeration   81 ( 949 )   908 - 912   2006年11月

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    記述言語:日本語   出版者・発行元:日本冷凍空調学会  

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  • Protease Susceptibility of β-Lactoglobulin Adsorbed on Stainless Steel Surface as Evidence of Contribution of Its Specific Segment to Adsorption

    Sakiyama Takaharu, Aya Atsuko, Embutsu Mari, IMAMURA Koreyoshi, NAKANISHI Kazuhiro

    Journal of bioscience and bioengineering   101 ( 5 )   434 - 439   2006年5月

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    記述言語:英語   出版者・発行元:公益社団法人日本生物工学会  

    β-Lactoglobulin (β-Lg) is a major constituent of fouling deposits in the dairy industry. To determine the interaction between β-Lg and stainless steel surfaces, β-Lg irreversibly adsorbed on stainless steel particles was subjected to lysyl endopeptidase treatment and the course of fragmentation was compared with that observed for β-Lg in solution. The results showed a distinct difference between the courses of fragmentation: a fragment (residues 102-135) was liberated readily from β-Lg in solution but scarcely from β-Lg irreversibly adsorbed on stainless steel particles. This result strongly suggests that residues 102-135 include a segment primarily responsible for the interaction of β-Lg with stainless steel surfaces. This supports our previous results [Sakiyama et al., J. Biosci. Bioeng., 88, 536-541 (1999)] that showed that residues 125-135 of β-Lg have a strong affinity toward stainless steel surfaces and probably a major contribution to the adsorption of β-Lg.

    DOI: 10.1263/jbb.101.434

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  • ポリスチレン表面に親和性を示すペプチドタグを付加した酵素の設計と特性解析

    今石大輔, 熊田陽一, 今中洋行, 今村維克, 崎山高明, 中西一弘

    日本農芸化学会中四国支部講演会講演要旨集   14th (Web)   2006年

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  • ポリスチレン親和性ペプチド標識タンパク質を用いる迅速免疫測定

    熊田陽一, 今中洋行, 今村維克, 加藤滋雄, 中西一弘

    化学工学会年会研究発表講演要旨集   71st   2006年

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  • Isolation and Characterization of a Thermostable Monoacylglycerol Lipase from Soil Bacterium

    Hiroyuki Imanaka, Masashi Mori, Koreyoshi Imamura, Takaharu Sakiyama, Kazuhiro Nakanishi

    Proceedings for Asia-Pacific Biochemical Engineering Conference (APBioChEC’04)   2005年

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  • 2E11-4 ポリスチレン親和性ペプチドを用いるタンパク質吸着の配向制御(生物化学工学,一般講演)

    熊田 陽一, 今中 洋行, 今村 維克, 加藤 滋雄, 崎山 高明, 中西 一弘

    日本生物工学会大会講演要旨集   17   153 - 153   2005年

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  • 糖類アモルファスマトリクスにおける糖-タンパク質間相互作用とタンパク質の安定化

    今村 維克

    低温生物工学会誌   51 ( 1 )   31 - 35   2005年

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    記述言語:日本語   出版者・発行元:低温生物工学会  

    Preserving the native conformation of protein during freeze-drying would lead to a greater stability against denaturation during storage. A number of investigators have reported that freeze-drying in the presence of sugar aids in preserving the native conformation of a protein. When a protein solution is freeze-dried in the presence of sugars, amorphous matrices of sugars are formed, which could embed protein molecules. The embedding of protein molecules in the amorphous sugar matrices is generally thought to prevent conformational changes in the protein during dehydration and storage due to interactions between protein and sugar molecules. In this review we describe the methods which are used to study the sugar-protein hydrogen bond. The effects of various sugars on the structural stabilization of protein in the dried state are also reviewed and discussed.

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  • ポリスチレン親和性ペプチド融合タンパク質の吸着特性とその応用

    徳永安秀, 熊田陽一, 今石大輔, 今中洋行, 今村維克, 崎山高明, 中西一弘

    化学工学会秋季大会研究発表講演要旨集(CD-ROM)   37th   2005年

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  • Effects of carboxyl groups on the adsorption behavior of low-molecular-weight substances on a stainless steel surface

    T Nagayasu, C Yoshioka, K Imamura, K Nakanishi

    JOURNAL OF COLLOID AND INTERFACE SCIENCE   279 ( 2 )   296 - 306   2004年11月

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    記述言語:英語   出版者・発行元:ACADEMIC PRESS INC ELSEVIER SCIENCE  

    The adsorption isotherms of various carboxylic acids and several amines on a stainless steel surface were taken as a function of pH and the ionic strength of the solution at 30 degreesC. In particular, the effect of the number of carboxyl groups on the adsorption behavior was investigated. Monocarboxylic acids such as benzoic, acid and n-butyric acid were reversibly adsorbed on the stainless steel particles and showed a Langmuir-type adsorption isotherm, i.e., Q = Kq(m)C/(1 + KC), where Q and C are, respectively, the amount of adsorbate adsorbed and the equilibrium concentration in the bulk solution, q(m), the maximum adsorbed amount, and K is the adsorption equilibrium constant. Carboxylic acids having plural carboxyl groups had much higher affinity to the surface and were adsorbed in both reversible and irreversible modes. The adsorption isotherms for the carboxylic acids having plural carboxyl groups could be expressed by a modified Langmuir-type adsorption isotherm, i.e., Q = q(irrev) + K q(rev)C/(1 + KC), where q(irrev) and q(rev) are, respectively, the maximum amounts adsorbed irreversibly and reversibly. The K and q(irrev) values increased with an increase in the number of carboxyl groups except for isophthalic acid and terephthalic acid. On the basis of the pH dependencies of K, q(m), q(irrev), and q(rev) as well as the surface properties of the stainless steel, both reversible and irreversible adsorptions were considered to occur through the electrostatic interaction between negatively charged carboxyl groups and the positively charged sites on the surface. The dependency of the qirrev value on ionic strength was discussed on the basis of the differences in their adsorbed state with the interaction forces to the surface and repulsive forces among the adsorbed molecules. The adsorption of amine components was quite weak. The RA-IR and molecular dynamics calculation were done to investigate the adsorption states of phthalic acid, trimellitic acid, and mellitic acid. (C) 2004 Elsevier Inc. All rights reserved.

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  • Characteristics of α-Glucosidase Production from Recombinant Aspergillus oryzae by Membrane-Surface Liquid Culture in Comparison with Various Cultivation Methods

    Morita Masakazu, Shimamura Hiroko, Ishida Natsuko, IMAMURA KOREYOSHI, SAKIYAMA TAKAHARU, NAKANISHI KAZUHIRO

    Journal of bioscience and bioengineering   98 ( 3 )   200 - 206   2004年9月

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    記述言語:英語   出版者・発行元:公益社団法人日本生物工学会  

    α-Glucosidase was produced using recombinant Aspergillus oryzae by membrane-surface liquid culture (MSLC), a method previously developed by the authors and the results compared with other methods, including shaking flask culture (SFC), agar-plate culture (APC), culture on urethane sponge supports (USC), and liquid surface culture (LSC) to determine possible reasons for the advantageous features of MSLC. When yeast extract was used as a nitrogen source, the amount of enzyme produced by MSLC was 5 or more times higher than those for SFC and LSC, but similar to that using APC. Enzyme production in USC was slightly lower than in MSLC and APC. Cell growth was similar irrespective of the cultivation method used. When NaNO_3, a typical inorganic nitrogen source was used, enzyme production in all the cultures was lower than that using yeast extract. However, even using NaNO_3, the amount of the enzyme produced by MSLC was 8 to 20 times higher than those by SFC, APC, USC, and LSC. Although cell growth using NaNO_3 was similar to that for yeast extract in MSLC, it was markedly decreased in SFC, APC, and LSC. The reason for the difference in enzyme productivity for various cultivation methods using yeast extract and NaNO_3 as a nitrogen source is discussed, on the basis of the experimental findings. The role of the oxygen transfer effect and gene expression levels in enzyme production were also examined.

    DOI: 10.1263/jbb.98.200

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  • Use of a novel affinity tag selected with a bacterial random peptide library for improving activity retention of glutathione S-transferase adsorbed on a polystyrene surface

    T Sakiyama, S Ueno, K Imamura, K Nakanishi

    JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC   28 ( 4-6 )   207 - 214   2004年6月

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    記述言語:英語   出版者・発行元:ELSEVIER SCIENCE BV  

    Aiming at developing a novel affinity tag for site-specific immobilization of functional proteins onto polystyrene (PS) surfaces, Escherichia coli random peptide display library was screened for dodecapeptides exhibiting a high affinity toward PS plate,. The selected peptides were commonly rich in hydrophobic amino acid residues and had two or three basic amino acid residues. Adsorption and desorption experiments for one of the selected peptide named PSI (KGLRGWREMISL) showed that it was well and irreversibly adsorbed onto PS latex particles. To study its performance as an affinity tag, PSI was genetically fused to a model enzyme, glutathione S-transferase (GST), in several manners, and the fusion enzymes were compared to the original GST in terms of the adsorption behavior onto the PS latex particles as well as the specific activities before and after the adsorption. The fusion GSTs in solution showed lower specific activities than the original one, and their adsorption behaviors were also altered. In particular, the fusion of PS 1 to the N-terminal region of GST resulted in severe losses both in the specific activity and in the adsorptive ability. However, two types of GSTs fused with PSI at the C-terminal region were well adsorbed onto the PS latex particles, and their specific activities after the adsorption were significantly higher than the original GST adsorbed on the PS latex particles. The fusion of PSI to the C-terminal region of GST was thus shown to reduce the activity loss upon the adsorption onto the PS latex particles. (C) 2004 Elsevier B.V. All rights reserved.

    DOI: 10.1016/j.molcatb.2003.12.019

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  • Production of nonproteinaceous amino acids using recombinant Escherichia coli cells expressing cysteine synthase and related enzymes with or without the secretion of O-acetyl-L-serine

    CH Zhao, K Ohno, K Sogoh, K Imamura, T Sakiyama, K Nakanishi

    JOURNAL OF BIOSCIENCE AND BIOENGINEERING   97 ( 5 )   322 - 328   2004年5月

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    記述言語:英語   出版者・発行元:SOC BIOSCIENCE BIOENGINEERING JAPAN  

    beta-(Pyrazol-1-yl)-L-alanine (beta-PA), a model nonproteinaceous amino acid, was specifically synthesized by two methods using recombinant Escherichia coli cells that express cysteine synthase, comprising serine acetyltransferase (SAT) and O-acetylserine sulfhydrylase-A (OASS-A) and related enzymes from E. coli. In the first method (method A), recombinant cells that express wildtype SAT, OASS-A, acetate kinase (AK), and phosphotransacetylase (PTA) showed the highest beta-PA production. beta-PA was produced at 140 mM from 200 MM L-serine and 200 mM pyrazole under optimum conditions. Using the cells expressing SATDeltaC20 (truncated SAT), OASS-A, AK, and PTA, beta-PA was produced at a level of only 80 mM, whereas O-acetyl-serine (OAS) was found to be secreted into the broth. Under optimum conditions, OAS accumulated at levels of around 105 mM from 300 mM L-serine. Thus, in the second method (method B), the secreted OAS was used as the substrate for the syntheses of beta-PA and beta-(triazol-1-yl)-L-alanine (beta-TA). The OAS that accumulated in the broth was efficiently converted to beta-PA and beta-TA at levels of around 90 mM from 105 mM OAS using free OASS-A. In both methods A and B, the addition of glucose was essential for the efficient production of beta-PA and OAS, respectively.

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  • Adsorption characteristics of bovine serum albumin and its peptide fragments on a stainless steel surface

    T Sakiyama, J Tomura, K Imamura, K Nakanishi

    COLLOIDS AND SURFACES B-BIOINTERFACES   33 ( 2 )   77 - 84   2004年1月

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    記述言語:英語   出版者・発行元:ELSEVIER SCIENCE BV  

    Adsorption characteristics of peptide fragments prepared by lysyl endopeptidase treatment of bovine serum albumin (BSA) were studied in comparison with those of BSA itself using stainless steel particles (type 316L) as a substrate surface. BSA was adsorbed onto the stainless steel surface remarkably at acidic pHs but scarcely at alkaline pHs. The peptide fragments were also scarcely adsorbed at alkaline pHs. At acidic pHs, however, the affinity toward the stainless steel surface depended on the type of peptide fragment; several types of the peptide fragments, relatively rich in acidic amino acid residues, such as DLGEEHFK (residues 13-20) and DAIPEDLPPLTADFAEDK (residues 295-312), were considerably adsorbed onto the stainless steel surface at acidic pHs. Similarly to BSA, the adsorption isotherms of those two types of peptide fragments showed very high affinities toward the stainless steel surface at pH 3 resulting in irreversible adsorption. Adsorption experiments for their synthetic analogues showed that the acidic amino acid residues were essential for the adsorption at pH 3. Furthermore FT-IR analysis suggested that the carboxyl groups of these acidic amino acid residues on the stainless steel surface were dissociated and hence had electrostatic interaction with the stainless steel surface. Thus the importance of carboxyl groups in the adsorption of peptide fragments on the stainless steel surface at acidic pHs was indicated. Close similarity of adsorption behaviors at acidic pHs found between BSA and those peptide fragments suggests that the acidic amino acid residues of the corresponding segments of a BSA molecule make a major contribution to the adsorption at acidic pHs. However, when those adsorbed at pH 3 were treated with 0.02 N NaOH, complete removal was observed for the peptide fragments but not for BSA. (C) 2003 Elsevier B.V. All rights reserved.

    DOI: 10.1016/j.colsurfb.2003.08.010

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  • ステンレス表面に対する低分子物質の吸着におけるカルボキシル基の寄与

    Journal of Colloid and Interface Science   279(2), pp. 289-295   2004年

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  • ランダムペプチドライブラリーから選別した新規アフィニティータグによるポリスチレン表面に吸着したグルタチオンSトランスフェラーゼの活性保持の改善

    J. Mol. Catal. B: Enzymatic   28 (4), pp. 207-214   2004年

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  • ステンレス表面に対する牛血清アルブミンとそのペプチド断片の吸着特性

    Colloids Surf. B: Biointerfaces   vol. 33(2), pp. 77-84   2004年

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  • システインシンセターゼおよび関連酵素を発現し,O-アセチルセリンを分泌能が異なる組み替え大腸菌を用いた非タンパク性アミノ酸の生産

    J. Biosci. Bioeng.   97(5), pp. 322-328   2004年

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  • Contribution of acidic amino residues to the adsorption of peptides onto a stainless steel surface

    K Imamura, Y Kawasaki, T Awadzu, T Sakiyama, K Nakanishi

    JOURNAL OF COLLOID AND INTERFACE SCIENCE   267 ( 2 )   294 - 301   2003年11月

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    記述言語:英語   出版者・発行元:ACADEMIC PRESS INC ELSEVIER SCIENCE  

    The role of the acidic amino acid residues in the adsorption of peptides/proteins onto stainless steel particles was investigated using a peptide fragment from bovine beta-lactoglobulin, Thr-Pro-Glu-Val-Asp-Asp-Glu-Ala-Leu-Glu-Lys (T5 peptide), which has a high affinity to a stainless steel surface at acidic pHs, and its mutant peptides substituted with different numbers of acidic amino acid residues. The adsorption behavior of the mutant peptides as well as the T5 peptide were studied at pH 3 with respect to concentration and ionic strength dependencies and the reversibility of the adsorption process. The behavior of the peptides was generally characterized as two distinct irreversible adsorption modes, Mode I and Mode II. In Mode I, the amounts adsorbed lay on the ordinate at zero equilibrium concentration in the solution, while in Mode II, the amount adsorbed increased with increased equilibrium concentration. The area occupied by the peptides was predicted by molecular mechanics and molecular dynamics. The state of the peptides, when adsorbed, was investigated using FT-IR analysis. The FT-IR analyses revealed that the side carboxylic groups of the peptides adsorbed on the stainless steel surface were ionized, while they were unionized in the solution at pH 3. Thus, the interactions between the carboxylic groups of the peptide and the stainless steel surface can be considered to be largely electrostatic. The peptide having four acidic amino acid residues took a maximum adsorbed amount, the reason for which is discussed. (C) 2003 Elsevier Inc. All rights reserved.

    DOI: 10.1016/S0021-9797(03)00700-8

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  • The kinetics of removal of proteins adsorbed on a stainless steel surface by H2O2-electrolysis and factors affecting its performance

    K Imamura, Watanabe, I, M Imada, T Sakiyama, K Nakanishi

    JOURNAL OF COLLOID AND INTERFACE SCIENCE   265 ( 1 )   49 - 55   2003年9月

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    記述言語:英語   出版者・発行元:ACADEMIC PRESS INC ELSEVIER SCIENCE  

    The kinetics of the removal of beta-lactoglobulin (beta-Lg) adsorbed on a stainless steel surface by H2O2-electrolysis treatment, in which hydroxyl radicals (OHs) generated by the electrolysis of hydrogen peroxide decompose the substances adhering on the surface, was investigated. The rate of removal of the adsorbed beta-Lg from the stainless steel surface during the treatment was monitored in situ by ellipsometry. The dependencies of the removal rate on the H2O2 concentration, the electric potential applied to the surface, and the supporting electrolyte concentration were examined and the results were compared with those obtained for the treatment of a titanium surface. Differences in the removal rates of the protein from the stainless steel and titanium surfaces are discussed with respect to differences in the nature of the interaction between the protein and the surface. The atomic compositions of the stainless steel surface before and after treatment were analyzed by Auger electron spectroscopy, and the stainless steel surface was found not to be affected by the H2O2-electrolysis treatment. The influences of various coexisting materials on removal characteristics during the H2O2-electrolysis treatment were also investigated. The difference between the effect of coexisting substances on the decomposition rate for the radical reaction in the H2O2-electrolysis treatment and that for the well-known UV-H2O2 treatment in bulk solution is discussed. (C) 2003 Elsevier Inc. All rights reserved.

    DOI: 10.1016/S0021-9797(03)00329-1

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  • 放線菌由来新規アシラーゼによるカプサイシン誘導体の合成とその抗酸化性の評価

    是石真友子, 張徳民, 川波和子, 今村維克, 崎山高明, 松村康生, 中西一弘

    日本食品工学会年次大会講演要旨集   4th   117   2003年7月

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  • 食品製造プロセスにおける新規洗浄方法の原理とその特性 (特集 食品工場の洗浄・除菌技術)

    今村 維克, 崎山 高明, 中西 一弘

    ジャパンフ-ドサイエンス   42 ( 6 )   27 - 33   2003年6月

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    記述言語:日本語   出版者・発行元:日本食品出版  

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  • 乾燥状態のタンパク質の安定性に及ぼす各種糖の効果

    Journal of Pharmacentical Sciences   92 ( 2 )   266 - 274   2003年2月

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  • Effects of types of sugar on the stabilization of protein in the dried state

    K Imamura, T Ogawa, T Sakiyama, K Nakanishi

    JOURNAL OF PHARMACEUTICAL SCIENCES   92 ( 2 )   266 - 274   2003年2月

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    記述言語:英語   出版者・発行元:JOHN WILEY & SONS INC  

    The effects of various sugars on the structural stabilization of protein during freeze-drying were investigated. The degree of native structure of protein that was freeze-dried and rehumidified at constant relative humidities (RHs) was evaluated by measurement of the alpha-helix content by Fourier-transform infrared spectroscopy. Bovine serum albumin (BSA) and several types of sugars, including sucrose, trehalose, and dextrans, were used as a model protein and sugars, respectively. The glass transition temperature, T-g for the amorphous sugar samples was measured by differential scanning calorimetry (DSC) to characterize the structural stability of sugars. The dependence of the alpha-helix content (Calpha-helix) of BSA on the sugar content (c(sugar)) could, in most cases, be represented by a Langmuir-type equation: Calpha-helix = K x (C-alpha-helix(max)-C-alpha-jelix(0)) x c(sugar)/(1 + K x c(sugar)) + C-alpha-helix(0), where K is a constant, indicating the ability of amorphous sugar matrix to embed protein, and C-alpha-helix(0) and C-alpha-helix(max) indicate the alpha-helix content in the absence of sugar and saturating levels of sugar, respectively. The preservation effects of the sugars could be characterized by K and C-alpha-helix(max). Both K and C-alpha-helix(max) values tended to be higher with decreasing T-g values for the amorphous sugar, probably because an amorphous sugar matrix with lower T-g values is structurally more flexible. The rehumidification of protein that was freeze-dried in the presence of sugar induced the refolding of protein structure, whereas the protein dried alone did not show any recovery of its native structure. (C) 2003 Wiley-Liss, Inc.

    DOI: 10.1002/jps.10305

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  • ペプチドのステンレス表面への吸着に対する酸性アミノ酸残基の寄与

    Journal of Colloid and Interface Science   276 ( 2 )   294 - 301   2003年

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  • ステンレス表面に吸着したタンパク質のH2O2-電気分解による脱離の速度論と処理効果に及ぼす諸因子の影響

    Journal of Colloid and Interface Science   265 ( 1 )   49 - 55   2003年

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  • Ionization characteristics of polyelectrolyte complex gels: analysis based on their swelling behaviors

    T. Sakiyama, T. Tsutsui, E. Masuda, K. Imamura, K. Nakanishi

    Macromolecules   36 ( 13 )   5039 - 5042   2003年

  • 酵素法による脂質付加食品素材物質の大量合成法の開発と機能性の解明

    中西 一弘, 今村 維克, 安達 修二

    年報   2003   271 - 277   2003年

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    記述言語:日本語   出版者・発行元:飯島記念食品科学振興財団  

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  • In situ analysis of the removal behavior of protein adsorbed on a titanium surface by H2O2-electrolysis treatment

    K Imamura, T Sakiyama, K Nakanishi

    LANGMUIR   18 ( 21 )   8033 - 8039   2002年10月

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    記述言語:英語   出版者・発行元:AMER CHEMICAL SOC  

    The kinetics of the removal of beta-lactoglobulin (beta-Lg) adsorbed to a titanium surface using a method involving the electrolysis of hydrogen peroxide (H2O2)-electrolysis treatment) was examined. The rate of decrease in the thickness of the layer of beta-Lg adsorbed to the titanium surface during the H2O2-electrolysis treatment was measured in situ by ellipsometry, The rate for this process followed first-order reaction kinetics. Relationships between the rate constant for removal and such factors as the H2O2 concentration, the potential applied to the titanium surface, the supporting electrolyte concentration, and temperature were experimentally studied. The rate of removal of the adsorbed beta-Lg from a stainless steel surface was found to be much slower than that from a titanium surface, indicating that the type of metal also affects the removal characteristics during the H2O2-electrolysis treatment.

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  • チタン表面に付着したタンパク質の過酸化水素-電気分解処理にする除去過程の直接解析

    Langmuir   18 ( 21 )   8033 - 8039   2002年10月

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  • Water sorption and glass transition behaviors of freeze-dried sucrose-dextran mixtures

    K Imamura, A Fukushima, K Sakaura, T Sugita, T Sakiyama, K Nakanishi

    JOURNAL OF PHARMACEUTICAL SCIENCES   91 ( 10 )   2175 - 2181   2002年10月

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    記述言語:英語   出版者・発行元:JOHN WILEY & SONS INC  

    The water sorption and glass transition behaviors of freeze-dried disaccharide-polysaccharide mixtures at various contents were investigated at relative humidities (RHs) of 0, 11, 23, and 33%. Sucrose and three types of dextrans, which differ in molecular weight, were used as model di- and polysaccharides, respectively. The relationship between the dextran and water contents of the sucrose-dextran mixture at different constant RHs indicated that a mixture of sucrose and dextran was lower than that calculated by the Lang and Steinberg mass balance equation. In the RH range of 0-23%, the glass transition temperature, T-g, increased to a considerable extent when the dextran content was equal to or higher than 50%, while the increase in T-g, at dextran contents lower than 50% was small. A marked increase in T-g, was observed at RH 33% for dextran contents of 0-25% as well as in the range above 50%. This suggests that the physical stability of the highly hydrated amorphous disaccharide is effectively strengthened by the addition of a small amount of polysaccharide. These tendencies were similar for the three dextrans of different molecular weights. Furthermore, it was demonstrated that the addition of a small amount of dextran is quite effective in preventing the collapse of amorphous sugar during freeze drying. (C) 2002 Wiley-Liss, Inc.

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  • 2.最近の洗浄・殺菌技術の動向 : 2.3 過酸化水素の電気分解を利用した新規洗浄方法

    今村 維克, 中西 一弘

    冷凍   77 ( 899 )   800 - 805   2002年9月

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    記述言語:日本語   出版者・発行元:日本冷凍空調学会  

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  • 粉末系におけるゼインの二次構造に及ぼす水分活性と脂質添加の影響

    水谷由記子, 松村康生, 今村維克, 中西一弘, 森友彦

    日本食品科学工学会大会講演集   49th   152   2002年8月

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  • 過酸化水素の電気分解によって発生する水酸化ラジカルを用いたタンパク質汚れの除去

    Journal of collord and Interface Sciences   250 ( 2 )   409 - 414   2002年6月

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  • Removal of proteinaceous soils using hydroxyl radicals generated by the electrolysis of hydrogen peroxide

    K Imamura, Y Tada, H Tanaka, T Sakiyama, K Nakanishi

    JOURNAL OF COLLOID AND INTERFACE SCIENCE   250 ( 2 )   409 - 414   2002年6月

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    記述言語:英語   出版者・発行元:ACADEMIC PRESS INC ELSEVIER SCIENCE  

    We have developed here for the first time a novel method to generate hydroxyl radicals, .OH, by applying slightly negative electric potentials (-0.2 - -0.8 V vs Ag/AgCl) to the surface of a metal (or metal oxide) that is in contact with hydrogen peroxide solution containing a supporting electrolyte. Namely, .OH radicals were generated at the surface by the electrolysis of hydrogen peroxide according to the equation, H2O2 + e(-) --&gt; .OH + OH-. This method was used to clean a stainless steel fouled with a model protein, beta-lactoglobulin. The .OHs generated at the surface were effective in removing beta-lactoglobulin that had been irreversibly adsorbed, by several minutes of treatment at room temperature (22 +/- 2degreesC). The removal rates measured for various concentrations of H2O2, and supporting electrolyte and different potentials were determined exclusively by the electric current. (C) 2002 Elsevier Science (USA).

    DOI: 10.1006/jcis.2002.8368

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  • 食品製造プロセスで発生する汚れの新規洗浄方法 (特集:食品製造装置の洗浄殺菌)

    中西 一弘, 崎山 高明, 今村 維克

    食品工業   45 ( 2 )   18 - 24   2002年1月

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    記述言語:日本語   出版者・発行元:光琳  

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  • メチレンブルーとその類似物質のステンレス表面への付着特性

    Journal of Colloid and Interface Science   245 ( 1 )   50 - 57   2002年1月

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  • Adsorption behavior of methylene blue and its congeners on a stainless steel surface

    K Imamura, E Ikeda, T Nagayasu, T Sakiyama, K Nakanishi

    JOURNAL OF COLLOID AND INTERFACE SCIENCE   245 ( 1 )   50 - 57   2002年1月

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    記述言語:英語   出版者・発行元:ACADEMIC PRESS INC ELSEVIER SCIENCE  

    Methylene blue and its congeners as model dyes were adsorbed onto stainless steel particles at different ionic strengths, pH values, and ethanol contents, and the adsorption mechanism was investigated. A Fourier transform infrared spectroscopy (FTIR) analysis of the dyes adsorbed on the stainless steel plate was carried out to determine the orientations of the adsorbed dyes on stainless steel surface. The adsorption isotherms for all the dyes tested were approximated by a Langmuir equation (Q = K q(m) C/(1 + KC)) in most cases except under strongly basic conditions. From the ionic strength and ethanol content dependencies of the K value in the Langmuir equations, both the electrostatic and hydrophobic interactions were indicated to contribute to the adsorption of the dyes at neutral pH. By comparing the K and q(m) values for the methylene blue congeners and with the aid of the FTIR analyses, it was found that the kind of substituent groups at most positions of the polyheterocycles of methylene blue strongly affects the adsorption behavior, particularly the area occupied by an adsorbed dye molecule, the affinity for the stainless steel surface, and the orientation of the adsorbed dye molecule on the stainless steel surface. (C) 2002 Elsevier Science.

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  • スクロースーデキストラン混合凍結乾燥材料の水分収着およびガラス転移特性

    Journal of pharmaceutical Sciences   91 ( 10 )   2175 - 2181   2002年

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  • サーモライシンの有機溶媒への懸濁方法が酵素活性に及ぼす影響

    田中 恵三, 今村 維克, 崎山 高明

    日本食品工学会誌   3 ( 1 )   15 - 23   2002年

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    記述言語:英語   出版者・発行元:日本食品工学会  

    DOI: 10.11301/jsfe2000.3.15

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  • S219 マイクロバイオリアクターとマイクロ生体反応素子の展望と課題(コンビナトリアル・バイオエンジニアリングの進展,シンポジウム)

    中西 一弘, 崎山 高明, 今村 維克

    日本生物工学会大会講演要旨集   14   21 - 21   2002年

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  • 308 酵素の固体表面に対する付着特性の解析と付着の制御(培養工学,一般講演)

    大西 宏輔, 今村 維克, 崎山 高明, 中西 一弘

    日本生物工学会大会講演要旨集   14   28 - 28   2002年

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  • 306 二糖-高分子複合アモルファスマトリクスの諸物性の解析(培養工学,一般講演)

    福島 厚志, 坂浦 啓介, 杉田 拓男, 今村 維克, 崎山 高明, 中西 一弘

    日本生物工学会大会講演要旨集   14   28 - 28   2002年

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  • 310 H_2O_<2^->電気分解処理における金属表面からのタンパク質汚れの除去(培養工学,一般講演)

    今村 維克, 渡辺 一平, 崎山 高明, 中西 一弘

    日本生物工学会大会講演要旨集   14   29 - 29   2002年

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  • 309 機能性タンパク質の固体表面への結合制御に関する基礎的研究(培養工学,一般講演)

    上野 俊輔, 崎山 高明, 今村 維克, 中西 一弘

    日本生物工学会大会講演要旨集   14   29 - 29   2002年

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  • 549 Streptomyces sp.由来新規アシラーゼの多様な基質特異性(酵素・酵素工学・タンパク質工学,一般講演)

    是石 真友子, 張 徳民, 高柳 栄子, 畑 和希, 今村 維克, 崎山 高明, 中西 一弘

    日本生物工学会大会講演要旨集   14   85 - 85   2002年

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  • Evaluation of hydration states of protein in freeze-dried amorphous sugar matrix

    K Imamura, M Iwai, T Ogawa, T Sakiyama, K Nakanishi

    JOURNAL OF PHARMACEUTICAL SCIENCES   90 ( 12 )   1955 - 1963   2001年12月

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    記述言語:英語   出版者・発行元:JOHN WILEY & SONS INC  

    A model to analyze the hydration state of protein in freeze-dried amorphous sugar matrix was proposed, based on the assumptions that there is a limit to the amount of protein that a given amount of amorphous sugar could embed and that the freeze-dried sugar-protein mixture is composed of the four components, i.e., sugars with and without hydrogen bonding to proteins and proteins with and without hydrogen bonding to sugar. Bovine serum albumin and three kinds of disaccharides, i.e., sucrose, maltose, and trehalose, were used as samples. Using the analytical equations derived from the model and experimental sugar content dependencies of the water sorption at various relative humidities, the amount of hydration water for bovine serum albumin, and the minimum amount of sugar to embed the protein were determined. On the basis of these results, the degree of interaction between the three sugars and protein was discussed, with respect to their stabilizing effect on the protein. (C) 2001 Wiley-Liss, Inc. and the American Pharmaceutical Association.

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  • Cleaning of a stainless steel surface fouled with protein using a UV-H2O2 technique

    K Imamura, Y Tada, H Tanaka, T Sakiyama, A Tanaka, Y Yamada, K Nakanishi

    JOURNAL OF CHEMICAL ENGINEERING OF JAPAN   34 ( 7 )   869 - 877   2001年7月

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    記述言語:英語   出版者・発行元:SOC CHEMICAL ENG JAPAN  

    The UV-H2O2 technique, which is commonly known as a method of decomposing organic substances from wastewater, was applied into cleaning of a stainless steel surface fouled with protein. On a stainless steel surface fouled with bovine beta -lactoglobulin (beta -Lg), H2O2 Solution was made to now, and UV rays were irradiated over the flowing liquid. The amounts of beta -Lg adsorbed before and during the UV-H2O2 cleaning were measured by a reflection absorption technique using Fourier transform infrared spectroscopy (RA-IR). The adsorbed amount approximately decreased linearly with time during the UV-H2O2 cleaning. There was an optimum H2O2 concentration for the removal rate. The H2O2 concentration dependency of the removal rate could be explained by considering the formation and disappearance rates of hydroxyl radicals (. OH), the decay of UV illuminance along the depth of the flow, and the rate of reaction between adsorbed beta -Lg and -OH.

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  • Increase in the stability of serine acetyltransferase from Escherichia coli against cold inactivation and proteolysis by forming a bienzyme complex

    K Mino, K Imamura, T Sakiyama, N Eisaki, A Matsuyama, K Nakanishi

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY   65 ( 4 )   865 - 874   2001年4月

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    記述言語:英語   出版者・発行元:TAYLOR & FRANCIS LTD  

    Cysteine synthetase from Escherichia coli is a bienzyme complex composed of serine acetyltransferase (SAT) and O-acetylserine sulfhydrylase-A, (OASS), The effects of the complex formation on the stability of SAT against cold inactivation and proteolysis were investigated. SAT was reversibly inactivated on cooling to 0 degreesC. Ultracentrifugal analysis showed that SET (a hexamer) was dissociated mostly into two trimers on cooling to 0 degreesC in the absence of OASS, while in the presence of BASS one trimer of the SAT subunits formed a complex with one dimer of BASS subunits. In the presence of BASS, trot only the cold inactivation rate was reduced but also the reactivation rate was increased. Furthermore, SAT became stable against proteolytic attack bg alpha -chymotrypsin and V8 protease by forming the complex with OASS, On the other hand, SAT was degraded by trypsin in the same manner both in the presence and in the absence of OASS. The different tendency in the stability against proteolysis with the different proteases was discussed with respect to the substrate specificity of the proteases and amino acid sequence of the C-terminal region of SAT that interacts with OASS.

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  • On the adsorption of proteins on solid surfaces, a common but very complicated phenomenon

    K Nakanishi, T Sakiyama, K Imamura

    JOURNAL OF BIOSCIENCE AND BIOENGINEERING   91 ( 3 )   233 - 244   2001年3月

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    記述言語:英語   掲載種別:書評論文,書評,文献紹介等   出版者・発行元:SOC BIOSCIENCE BIOENGINEERING JAPAN  

    Adsorption of proteins on solid surfaces and their interaction are major concerns in a number of fields such as biology, medicine, biotechnology and food processing, and play an important role from various points of view. Based on practical viewpoints, information on the conformation of the adsorbed protein as well as adsorption characteristics is essential for a system's performance. Although there are still many problems to be solved, extensive studies in recent years, owing to the development in instrumentation and instrumental techniques, reveal the adsorption behavior of proteins in detail. Here, we stress the importance and interesting aspect of protein adsorption on solid surfaces by reviewing findings that have been obtained in recent years.

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  • 凍結乾燥によって得られた糖類アモリファスマトリクスにおけるタンパク質の水和状態の評価

    Journal of Pharmacentical Sciences   91(12), 1955-1964   2001年

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  • UV/H2O2処理を利用したステンレス表面に付着したタンパク質の洗浄

    34 ( 7 )   869 - 877   2001年

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  • 413 膜面液体培養法を用いた遺伝子組換えAspergillus oryzaeによる酵素生産

    島村 寛子, 森田 正和, 与那城 亮太, 今村 維克, 崎山 高明, 峰時 俊貴, 中西 一弘

    日本生物工学会大会講演要旨集   13   199 - 199   2001年

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  • 246 トランスグルタミナーゼによるβ-ラクトグロブリンの脂肪酸修飾

    和田 栄子, 増田 治樹, 今村 維克, 崎山 高明, 安達 修二, 松野 隆一, 中西 一弘

    日本生物工学会大会講演要旨集   13   158 - 158   2001年

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  • 247 Streptomyces sp.由来カプサイシン加水分解酵素の精製

    是石 真友子, 張 徳民, 今村 維克, 崎山 高明, 安達 修二, 松野 隆一, 中西 一弘

    日本生物工学会大会講演要旨集   13   159 - 159   2001年

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  • 655 キトサン/コンドロイチン硫酸複合ゲルのpH応答型膨潤特性とその応用

    崎山 高明, 生田 祐嗣, 今村 維克, 中西 一弘

    日本生物工学会大会講演要旨集   13   282 - 282   2001年

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  • 656 H_2O_2-電気分解処理によるステンレス表面に付着したタンパク質汚れの除去

    今村 維克, 多田 洋一郎, 田中 啓文, 崎山 高明, 中西 一弘, 田中 淳已, 山田 要輔

    日本生物工学会大会講演要旨集   13   282 - 282   2001年

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  • バイオ・食品製造プロセスにおける汚れの付着と洗浄 (特集 最近の生物工学の展望)

    中西 一弘, 崎山 高明, 今村 維克

    ケミカルエンジニヤリング   45 ( 11 )   854 - 861   2000年11月

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    記述言語:日本語   出版者・発行元:化学工業社  

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  • ステンレス表面への各種アミノ酸の吸着特性

    Journal of collorid and Interface Science   229 ( 1 )   237 - 246   2000年9月

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  • Characteristics of serine acetyltransferase from Escherichia coli deleting different lengths of amino acid residues from the C-terminus

    K Mino, K Hiraoka, K Imamura, T Sakiyama, N Eisaki, A Matsuyama, K Nakanishi

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY   64 ( 9 )   1874 - 1880   2000年9月

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    記述言語:英語   出版者・発行元:TAYLOR & FRANCIS LTD  

    Some properties of serine acetyltransferases (SATs) from Escherichia coli, deleting 10-25 amino acid residues from the C-terminus (SAT Delta C10-Delta C25) were investigated. The specific activity depended only slightly on the length of the C-terminal region deleted. Although the sensitivity of SAT Delta C10 to inhibition by L-cysteine was similar to that for the wild-type SAT, it became less with further increases in the length of the amino acid residues deleted. SAT Delta C10 was inactivated on cooling to 0 degrees C and dissociated into dimers or trimers in the same manner as the wild-type SAT, but Met-256-Ile mutant SAT as web as SAT Delta C14, SAT Delta C20, and SAT Delta C25 were stable. Since SAT Delta C10, SAT Delta C14, and SAT Delta C25 did not form a complex with O-acetyl-serine sulfhydrylase-A (OASS-A) in a way similar to SAT Delta C20. it was indicated that 10 amino acid residues or fewer from the C-terminus of the wild-type SAT are responsible for the complex formation with OASS-A. The C-terminal peptide of the 10 amino acid residues interacted competitively with OASS-A with respect to OAS although its affinity was much lower than that for the wild-type SAT.

    DOI: 10.1271/bbb.64.1874

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  • Adsorption behavior of amino acids on a stainless steel surface

    K Imamura, T Mimura, M Okamoto, T Sakiyama, K Nakanishi

    JOURNAL OF COLLOID AND INTERFACE SCIENCE   229 ( 1 )   237 - 246   2000年9月

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    記述言語:英語   出版者・発行元:ACADEMIC PRESS INC  

    The adsorption behavior of various amino acids on a stainless steel surface was investigated at 30 degrees C and over a pH range of 3-10. Acidic and basic amino acids except histidine adsorbed remarkably at pH 3-4 and 7-10, respectively, and showed Langmuir-type adsorption isotherms. The effects of pH and ionic strength on the adsorption isotherms were investigated to analyze the interactions between amino acids and adsorption sites on the stainless steel. Hydrophobic amino acids and glycine showed only small adsorbed amounts at all pHs tested. For the acidic and basic amino acids, reversibility of the absorption and the influence of the ionic strength on the adsorption behavior were examined. The adsorption isotherms of the derivatives of aspartic acid were also measured in order to examine the contribution of the carboxylic groups of acidic amino acids to the adsorption. Furthermore, a Fourier-transform infrared spectroscopic analysis and semiempirical molecular orbital calculation were carried out to analyze the ionization states and the configuration of the amino acids adsorbed on a stainless steel surface. These investigations suggest that the acidic and basic amino acids adsorb through two electrostatic interactions of two ionized groups in the amino acid with a stainless steel surface, (C) 2000 Academic Press.

    DOI: 10.1006/jcis.2000.7016

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  • Water sorption and glass transition of amorphous sugars containing BSA

    K Imamura, T Suzuki, T Tatsumichi, S Kirii, M Okazaki

    JOURNAL OF CHEMICAL ENGINEERING OF JAPAN   33 ( 4 )   657 - 660   2000年8月

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    記述言語:英語   出版者・発行元:SOC CHEMICAL ENG JAPAN  

    Water sorption and glass transition of four amorphous sugars (lactose, maltose, sucrose, and trehalose) containing bovine serum albumin (BSA) are investigated. Freeze-dried sugar-BSA samples equilibrated at several water activities ranging from 0 to 0.43 were prepared. Moisture content and glass transition temperature (T-g) were measured. For the all sugars, it is found that BSA lowers T-g at low water activity, and raises it at high water activity. It Is also found that the difference between T-g of the sugar-BSA samples and that of the corresponding amorphous sugar samples (T-go) depends mainly on T-go.

    DOI: 10.1252/jcej.33.657

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  • Kinetics and equilibrium for thermolysin-catalyzed syntheses of dipeptide precursors in aqueous/organic biphasic systems

    M Miyanaga, M Ohmori, K Imamura, T Sakiyama, K Nakanishi

    JOURNAL OF BIOSCIENCE AND BIOENGINEERING   90 ( 1 )   43 - 51   2000年7月

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    記述言語:英語   出版者・発行元:SOC BIOSCIENCE BIOENGINEERING JAPAN  

    The initial kinetics for the syntheses of N-(benzyloxycarbonyl)-L-alanyl-L-phenylanine methyl ester (ZAPM) and N-(benzyloxycarbonyl)-L-aspartyl-L-phenylalanine methyl ester (ZDPM) in an aqueous/organic biphasic system, using free thermolysin were elucidated, both experimentally and theoretically. As model organic solvents, ethyl acetate and tert-amyl alcohol were used. The substrate concentration dependencies of the initial rate of syntheses for ZAPM and ZI)PM observed in the biphasic system were well simulated using the overall partition coefficients of the substrates and product taking into consideration the effect of the formation of ion-pair complexes between the acid and amine components of the substrate, the initial rate equations determined in an aqueous buffer saturated with the organic solvent, and the pH dependence of the rate constant. The equilibrium yield for the synthesis of ZDPM was also in good agreement with the calculated result using the overall partition coefficients and equilibrium constant measured in the aqueous buffer.

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  • Kinetic analysis for synthesis of a dipeptide precursor using an immobilized enzyme in water-immiscible organic solvents

    M Miyanaga, K Imamura, T Sakiyama, K Nakanishi

    JOURNAL OF BIOSCIENCE AND BIOENGINEERING   90 ( 1 )   112 - 114   2000年7月

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    記述言語:英語   出版者・発行元:SOC BIOSCIENCE BIOENGINEERING JAPAN  

    N-(Benzyloxycarbonyl)-L-aspartyl-L-phenylalanine methyl ester (Z-AspPheOMe), a precursor of the synthetic sweetener aspartame, was synthesized, using thermolysin immobilized onto Amberlite XAD-7, both in ethyl acetate and in tert-amyl alcohol. The initial rates for synthesis of Z-AspPheOMe in the organic solvents were predicted on the basis of a model proposed for an aqueous/organic biphasic reaction and compared with the experimentally observed substrate concentration dependencies. The experimental synthetic rates using the enzyme immobilized at a high enzyme concentration were lower than the calculated ones over a wide range of the substrate concentration. It was suggested as a reason for this discrepancy that the enzyme molecules form compact aggregates and those existing inside the aggregates cannot be utilized for reaction. The experimental results with the enzyme immobilized at a low concentration in ethyl acetate coincided well with the calculated ones. On the other hand, when tert-amyl alcohol was used, the experimental results were different in tendency irrespective of the amount of enzyme loaded, probably due to the fact that a distinct water phase does not exist around the enzyme aggregates inside the support.

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  • Development of novel advanced oxidation system using combined UV/H2O2 technique and kinetic analysis for decomposition of dye solutions

    K Imamura, A Hiramatsu, M Imada, T Sakiyama, A Tanaka, Y Yamada, K Nakanishi

    JOURNAL OF CHEMICAL ENGINEERING OF JAPAN   33 ( 2 )   253 - 261   2000年4月

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    記述言語:英語   出版者・発行元:SOC CHEMICAL ENG JAPAN  

    A novel advanced oxidation system using a combined UV/H2O2 technique was constructed for application to wastewater treatment. In this system, wastewater containing H2O2 flows along a channel with a flat surface in a thin film, onto which surface UV rays with a wavelength of 253.7 nm are irradiated. As a model wastewater, we used various dye solutions and investigated decoloration rates under various conditions differing in H2O2 concentration, temperature, UV illuminance, and so on. Based on the model, which takes into consideration the formation as well as the disappearance rate of hydroxyl radical (. OH), distribution of UV illuminance along the depth of the flow, and the rate of reaction between dye and . OH, we could simulate the course of dye decomposition. Furthermore, we showed the existence of an optimum H2O2 concentration for decomposition on the basis of this model.

    DOI: 10.1252/jcej.33.253

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  • 膜面液体培養法を用いた海洋植物プランクトンOscillatoria属の培養と特性解明

    中西 一弘, 崎山 高明, 今村 維克

    財団法人ソルト・サイエンス研究財団助成研究報告集   1998 ( 1 )   279 - 286   2000年3月

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    記述言語:日本語   出版者・発行元:ソルト・サイエンス研究財団  

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  • BSAを含有した糖アモルファスの水分収着およびガス転移

    Journal of Chemical Engineering of Japan   33 ( 4 )   657 - 660   2000年

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  • 紫外線/H2O2処理を利用した新規促進酸化システムの開発と色素水溶液の分解過程の速度論の解析

    33 ( 2 )   253 - 261   2000年

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  • 942 複合体形成によるEscherichia coli由来セリンアセチルトランスフェラーゼの低温失活及びプロテアーゼに対する安定性の増加

    三野 光識, 今村 維克, 崎山 高明, 永崎 直樹, 松山 旭, 中西 一弘, RITE

    日本生物工学会大会講演要旨集   12   225 - 225   2000年

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  • 927 アミノアシラーゼを用いたカプサイシン誘導体の合成

    是石 真友子, 鎌倉 薫, 今村 維克, 崎山 高明, 安達 修二, 松野 隆一, 中西 一弘

    日本生物工学会大会講演要旨集   12   218 - 218   2000年

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  • 929 水可溶性溶媒中でのN-アシルアミノ酸の酵素合成

    和田 栄子, 半田 将人, 今村 維克, 崎山 高明, 安達 修二, 松野 隆一, 中西 一弘

    日本生物工学会大会講演要旨集   12   219 - 219   2000年

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  • 928 アセチルCoA再生系を組み込んだシステイン合成酵素を用いる非天然アミノ酸合成

    大野 克博, 三野 光識, 今村 維克, 崎山 高明, 永崎 直樹, 松山 旭, 中西 一弘

    日本生物工学会大会講演要旨集   12   218 - 218   2000年

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  • 1137 糖類アモルファスによる乾燥タンパク質の構造保持作用

    今村 維克, 小川 智広, 崎山 高明, 中西 一弘

    日本生物工学会大会講演要旨集   12   280 - 280   2000年

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  • Analysis for partition equilibrium of amino acid derivatives in aqueous/organic biphasic systems

    M Miyanaga, K Imamura, K Tanaka, T Sakiyama, K Nakanishi

    JOURNAL OF BIOSCIENCE AND BIOENGINEERING   88 ( 6 )   651 - 658   1999年12月

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    記述言語:英語   出版者・発行元:SOC BIOSCIENCE BIOENGINEERING JAPAN  

    The overall partition coefficients of the acid and amine components of amino acid derivatives, the substrates for proteinase-catalyzed synthesis of oligopeptide precursors, in an aqueous/organic biphasic system were studied both experimentally and theoretically. In a single-component system containing either an acid or amine component, the overall partition coefficient was well expressed by a model using the acid dissociation constant and partition coefficients for the non-ionized and ionized forms of the component. The overall partition coefficient in a binary-component system containing both acid and amine components was well simulated by the model when the formation of ion-pair complexes in the organic solvent phase was taken into consideration in addition to the partitioning of the non-ionized and ionized forms of the components. The formation of ion-pair complexes in the organic solvent phase was indicated by an analysis using Fourier transform infrared ray spectroscopy (FT-IR). In addition to the partition equilibrium, the aqueous-phase pH change after partitioning could be predicted by the model.

    DOI: 10.1016/S1389-1723(00)87095-4

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  • Adsoption Characteristics of Tryptic Fragments of Bovine β-Lactoglobuilin on a Stainless Steel Surface

    Sakiyama Takaharu, Tanino Kei, Urakawa Masako, IMAMURA Koreyoshi, TAKAHASHI Tokio, NAGAI Takashi, NAKANISHI Kazuhiro

    Journal of bioscience and bioengineering   88 ( 5 )   536 - 541   1999年11月

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    記述言語:英語   出版者・発行元:公益社団法人日本生物工学会  

    As a strategy for the analysis of the mode of protein adsorption onto stainless steel surfaces, peptides obtained by tryptic digestion of bovine β-lactoglobulin were subjected to adsorption experiments after identification of their primary structures. In the presence of 1 mM KOH, the peptides were scarcely adsorbed onto the surfaces of stainless steel particles from the peptide mixture. The adsorption experiments on isolated peptides showed that the affinities of the peptides for stainless steel surfaces in the presence of 1 mH HNO_3 were significantly different from each other. Peptides without any acidic amino acid residues were scarcely adsorbed onto the surface, whereas some peptides with acidic amino acid residues were found to be irreversibly adsorbed onto the surfaces in the acidic pH region. As for the latter peptides, the amount adsorbed on the surface increased with increasing ionic strength. These results indicated that the carboxyl groups on the side chains of the peptides play an important role in the adsorption. Furthermore, the adsorption behavior of β-lactoglobulin itself was found to be very similar to that of one of the latter peptides.

    DOI: 10.1016/S1389-1723(00)87672-0

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  • Stability of immobilized thermolysin in organic solvents

    M Miyanaga, M Ohmori, K Imamura, T Sakiyama, K Nakanishi

    JOURNAL OF BIOSCIENCE AND BIOENGINEERING   87 ( 4 )   463 - 472   1999年4月

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    記述言語:英語   出版者・発行元:SOC BIOSCIENCE BIOENGINEERING JAPAN  

    Various factors affecting the stability of thermolysin immobilized by cross-linking with glutaraldehyde were elucidated, particularly in the water-immiscible organic solvents such as ethyl acetate and tert-amyl alcohol. The main reason for enzyme inactivation in water-immiscible organic solvents was found to be autolysis in the water phase, which may surround the enzyme immobilized inside the support. By contrast, in water-miscible organic solvents thermal denaturation was the predominant cause of enzyme inactivation. Courses of inactivation were expressed by second-order kinetics in the initial stage, after which inactivation proceeded at a slower rate. The extent of autolysis was found to strongly depend on the kind of organic solvent, the water content, and type of support and these dependencies were explained by the difference in the amount and state of water inside the support. Thermolysin was immobilized onto Amberlite XAD-7 as a compact aggregate inside the support which may increase the stability of the enzyme. Finally, it was shown that the stability of the immobilized enzyme could be correlated with the logP value for water-miscible organic solvents and with the solubility of water for water- immiscible organic solvents.

    DOI: 10.1016/S1389-1723(99)80095-4

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  • 食品製造・バイオプロセスにおける洗浄とタンパク性汚れの付着(<特集>殺菌・洗浄のバイオテクノロジー)

    中西 一弘, 今村 維克, 崎山 高明

    生物工学会誌 : seibutsu-kogaku kaishi   77 ( 6 )   241 - 245   1999年

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    その他リンク: http://dl.ndl.go.jp/info:ndljp/pid/10513378

  • 1018 膜面液体培養法(MSLC)を用いたAspergillue oryzae NRRL 484によるコウジ酸生産と特性解明

    杉田 拓男, 瀬川 貴之, 島村 寛子, 今村 維克, 崎山 高明, 中西 一弘

    日本生物工学会大会講演要旨集   11   219 - 219   1999年

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  • 822 大腸菌由来システイン合成酵素の特性と速度論的解析

    三野 光識, 今村 維克, 崎山 高明, 永崎 直樹, 松山 旭, 中西 一弘, RITE

    日本生物工学会大会講演要旨集   11   170 - 170   1999年

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  • 1421 タンパク質・ペプチド・アミノ酸のステンレス表面への付着特性の解析

    今村 維克, 佐藤 昌子, 戸村 淳嗣, 岡本 真琴, 崎山 高明, 中西 一弘, 高橋 時夫, 長井 直士

    日本生物工学会大会講演要旨集   11   333 - 333   1999年

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  • 1019 膜面液体培養(MSLC)を用いたMonascus pilosus kによる色素生産

    津嶋 容子, 味野 晃, 今村 維克, 崎山 高明, 中西 一弘, 菅 貞治, 段王 保文

    日本生物工学会大会講演要旨集   11   220 - 220   1999年

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  • 817 C-末端欠失セリンアセチルトランスフェラーゼの精製と諸性質

    平岡 憲二, 三野 光識, 今村 維克, 崎山 高明, 永崎 直樹, 松山 旭, 中西 一弘, RITE

    日本生物工学会大会講演要旨集   11   167 - 167   1999年

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  • 850 アセチルCoA再生系を組込んだシステインシンテターゼを用いたβ-ピラゾールアラニンの合成

    大野 克博, 三野 光識, 山野上 豪, 今村 維克, 崎山 高明, 永崎 直樹, 松山 旭, 中西 一弘, RITE

    日本生物工学会大会講演要旨集   11   184 - 184   1999年

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  • 1020 膜面液体培養(MSLC)を用いた遺伝子組換えAspergillus oryzaeによる酵素生産と特性解明

    島村 寛子, 石田 奈津子, 杉田 拓男, 今村 維克, 崎山 高明, 中西 一弘, 尾関 健二

    日本生物工学会大会講演要旨集   11   220 - 220   1999年

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  • Influence of protein on phase transition of amorphous sugar

    K Imamura, T Suzuki, S Kirii, T Tatsumichi, M Okazaki

    JOURNAL OF CHEMICAL ENGINEERING OF JAPAN   31 ( 3 )   325 - 329   1998年6月

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    記述言語:英語   出版者・発行元:SOC CHEMICAL ENG JAPAN  

    In order to investigate the influence of protein on the phase transitions of amorphous structure of sugar, we carried out differential scanning calorimetry (DSC) analysis of freeze-dried sucrose containing various contents of water and bovine serum albumin (BSA). Glass transition temperature Tg, crystallization temperature Tcr, and melting temperature Tm were measured. Tg, Tcr, and Tm decreased with increasing water activity. Addition of BSA raised Tg except the case that water activity was low (up to about 0.1). Tcr increased linearly with BSA content, and Tm did not depend on BSA content. The increase in Tg and Tcr with BSA suggests that protein contributes to stabilizing amorphous structure of sugar.

    DOI: 10.1252/jcej.31.325

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  • Fouling and cleaning of stainless steel surface: Adsorption and desorption behavior of bovine serum albumin and gelatin

    T Sakiyama, T Toyomasu, A Nagata, K Imamura, K Nakanishi, T Takahashi, T Nagai

    JOURNAL OF CHEMICAL ENGINEERING OF JAPAN   31 ( 2 )   208 - 213   1998年4月

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    記述言語:英語   出版者・発行元:SOC CHEMICAL ENG JAPAN  

    Adsorption and desorption behavior of bovine serum albumin (BSA) and gelatin on the surface of stainless steel particles are studied. The amount of BSA adsorbed increases significantly with temperature above 60 degrees C whereas that of gelatin decreases slightly with increasing temperature, Results of adsorption experiments with S-carboxymethylated BSA show that the thermal aggregation of BSA molecules at the surface through intermolecular thiol-disulfide interchange reactions plays an important role in the adsorption of BSA at elevated temperatures. Furthermore, the initial desorption rate constants and residual amounts of the two proteins during caustic and enzymatic cleanings of the fouled particles are compared under various conditions. As a result, a large difference is found in the temperature dependence of the initial desorption rate constant in caustic cleaning, suggesting different modes of adsorption of the proteins.

    DOI: 10.1252/jcej.31.208

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  • Performance of protease as a cleaning agent for stainless steel surfaces fouled with protein

    T Sakiyama, T Toyomasu, A Nagata, K Imamura, T Takahashi, T Nagai, K Nakanishi

    JOURNAL OF FERMENTATION AND BIOENGINEERING   85 ( 3 )   297 - 301   1998年

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    記述言語:英語   出版者・発行元:SOC FERMENTATION BIOENGINEERING, JAPAN  

    Cleaning experiments were performed by feeding protease solutions as cleaning agents into a packed column of stainless steel particles fouled with beta-lactoglobulin or gelatin. By evaluating the initial desorption rate constants (Brst-order rate constants) and residual amounts of protein, the cleaning efficiencies of the proteases were compared. The initial desorption rate constant depended on the kind of protease used and also on the type of protein to be removed. It increased with protease concentration and reached a constant value at a limit concentration. Kinetic analysis of the proteolytic reactions catalyzed by the proteases revealed that protease with a large Vm(ax)/EKm value for the proteolytic reaction resulted in a large value of the initial desorption rate constant in the enzymatic cleaning at a low protease concentration. The K-m value affected the limit concentration of protease yielding a saturation of initial desorption rate constant.

    DOI: 10.1016/S0922-338X(97)85678-4

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  • Development of a cylindrical apparatus for membrane-surface liquid culture and production of kojic acid using Aspergillus oryzae NRRL484

    Y Wakisaka, T Segawa, K Imamura, T Sakiyama, K Nakanishi

    JOURNAL OF FERMENTATION AND BIOENGINEERING   85 ( 5 )   488 - 494   1998年

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    記述言語:英語   出版者・発行元:SOC FERMENTATION BIOENGINEERING, JAPAN  

    To apply membrane-surface liquid culture (MSLC), reported in previous papers (J. Ferment. Bioeng., 80: 35-40, 41-46, 1995) to large-scale production, a novel bioreactor was constructed in which molds were grown on the outer surface of a cylindrical porous membrane facing the air with liquid medium running down along its inner surface. We cultivated Aspergillus oryzae NRRL484 to produce kojic acid as a model secondary metabolite. An SE20 membrane composed of polysulfone with a nominal pore diameter of 0.2 mu m (Fuji Photo Film Co.) was found to be most suitable based on the facts that the mass transfer through the membrane had no appreciable effect on the kojic acid production and the mycelia did not pass through the membrane pores. Kojic acid was produced at higher levels than in shaking flask culture. The highest kojic acid concentration in the batch MSLCs using the cylindrical membrane module with an active surface area of 220 cm(2) was 14, 45, and 60 mg/ml for glucose concentrations in the medium of 5, 10, and 20%, respectively. In the shaking flask cultures using 10 and 20% glucose, the maximum kojic acid concentrations were 24 and 22 mg/ml, respectively. In the MS;LC, the liquid medium was quite clear without any contamination of cells. Furthermore, the mold was highly stable against autolysis, and continuous cultivation with holding tells on the membrane surface could be conducted for over 70 d. When the average feed rate of the medium containing 10% glucose and 0.1% yeast extract was 30 ml/d, the steady-state kojic acid concentration at the outlet of the MSLC apparatus was in the range of 45-50 mg/ml.

    DOI: 10.1016/S0922-338X(98)80067-6

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  • 糖類アモルファス構造の相転移に及ぼすタンパク質の影響

    Journal of Chemical Engineering of Japan   31 ( 3 )   325 - 329   1998年

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    記述言語:英語  

    DOI: 10.1252/jcej.31.325

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  • 新規に開発したカビの膜面液体培養法を用いた食品加工用酵素の生産に関する基礎研究

    中西 一弘, 崎山 高明, 今村 維克

    食に関する助成研究調査報告書   ( 11 )   51 - 58   1998年

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    記述言語:日本語   出版者・発行元:すかいら-くフ-ドサイエンス研究所  

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  • 324 膜面液体培養法によるカビの培養とそのスケールアップ

    瀬川 貴之, 今村 維克, 崎山 高明, 中西 一弘

    日本生物工学会大会講演要旨集   9   96 - 96   1997年

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    記述言語:日本語   出版者・発行元:日本生物工学会  

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  • 540 アミノ酸およびペプチド類のステンレス粒子への吸脱着挙動

    三村 朋也, 谷野 兄, 今村 維克, 崎山 高明, 中西 一弘

    日本生物工学会大会講演要旨集   9   190 - 190   1997年

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    記述言語:日本語   出版者・発行元:日本生物工学会  

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講演・口頭発表等

  • 噴霧乾燥による粒子状〜ナノ繊維状アモルファス固体分散の調製と特性評価

    佐藤 春奈, 本田 奈央, 長谷川 高飛, 荒賀 智佳, 今中 洋行, 石田 尚之, 今村 維克

    化学工学会第55回秋季大会  2024年9月13日 

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    開催年月日: 2024年9月11日 - 2024年9月13日

    記述言語:日本語   会議種別:ポスター発表  

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  • 温度走査フーリエ変換赤外分光分析による温度応答性ポリマーの水-アルコール混合溶媒中での構造変化の解析

    武澤 歩, 石田 尚之, 今中 洋行, 今村 維克

    化学工学会第55回秋季大会  2024年9月12日 

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    開催年月日: 2024年9月11日 - 2024年9月13日

    記述言語:日本語   会議種別:ポスター発表  

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  • 湿式および乾式紡糸による多糖ファイバーの調製と物性評価

    今村 維克, 福嶋 海人, 佐藤 春奈, 荒賀 智佳, 今中 洋行, 石田尚之

    2024年9月12日 

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    開催年月日: 2024年9月11日 - 2024年9月13日

    記述言語:日本語   会議種別:口頭発表(一般)  

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  • タンパク質性リガンドの保存安定性:固定化後の相互作用検出感度評価

    渡邉 滉大, 今村 維克, 今中 洋行

    化学工学会第55回秋季大会  2024年9月11日 

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    開催年月日: 2024年9月11日 - 2024年9月13日

    記述言語:日本語   会議種別:ポスター発表  

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  • 分子構造が異なる香料のアモルファス固体分散による被包括特性の比較

    長谷川高飛, 本田奈央, 佐藤春奈, 今中洋行, 石田尚之, 今村維克

    日本食品工学 会 第 2 5 回(2024年度)年次大会  2024年8月20日 

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    開催年月日: 2024年8月19日 - 2024年8月20日

    記述言語:日本語   会議種別:口頭発表(一般)  

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  • 二流体ノズル式噴霧乾燥による薬剤キャリア物質の ナノファイバー形成条件の探索

    本田奈央, 荒賀智佳, 長谷川高飛, 佐藤春奈, 今中洋行, 今村維克

    日本薬剤学会第39年会  2024年5月24日 

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    開催年月日: 2024年5月23日 - 2024年5月25日

    記述言語:日本語   会議種別:ポスター発表  

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  • 噴霧乾燥による両親媒性高分子からなるサブミクロン繊維の 調製と難水溶性物質キャリアとしての応用

    佐藤春奈, 本田奈央, 長谷川高飛, 荒賀智佳, 今中洋行, 今村維克

    日本薬剤学会第39年会  2024年5月24日 

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    開催年月日: 2024年5月23日 - 2024年5月25日

    記述言語:日本語   会議種別:口頭発表(一般)  

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  • 金属表面の高分子コーティングによるタンパク質の三次元固定化

    田坂 隆貴, 西川 葵, 今中 洋行, 石田 尚之, 今村 維克

    化学工学会岡山大会2022  2022年12月19日 

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    開催年月日: 2022年12月19日

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受賞

  • 研究賞

    2019年8月   日本食品工学会   食品タンパク質をはじめとする生体分子の包括安定化技術に関する工学研究

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  • 奨励賞

    2007年8月   日本食品工学会   過酸化水素の電気分解を利用した金属表面の新規洗浄技術の開発とその高度化に関する研究

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  • 奨励賞

    2002年   化学工学会   水酸化ラジカルを利用したバイオプロセスの高度洗浄に関する基礎的研究

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    受賞国:日本国

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共同研究・競争的資金等の研究

  • Surfactant-free固体分散の物理的安定化と難水溶性薬剤の水溶性改善

    研究課題/領域番号:19H02499  2019年04月 - 2023年03月

    日本学術振興会  科学研究費助成事業  基盤研究(B)

    今村 維克

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    配分額:16120000円 ( 直接経費:12400000円 、 間接経費:3720000円 )

    難水溶性薬剤の生体内における溶解性を改善する手法として,水可溶性のキャリアマトリクスに非晶質化した薬剤を分散する技術がある.申請者は,本来は“水と油”の関係にある糖と疎水性薬剤を,両親媒性物質を一切用いることなく,分子レベルで均一に混合できる原理的に新規な固体分散技術(Sole Amorphous Sugar based Amorphous Solid Dispersioin, SAS-SD, “surfactant-free固体分散”から呼称変更)を開発した.このSAS-SD試料において糖は水中で瞬時に溶解するため,疎水性薬剤の対水溶解性を既存の固体分散技術と同等以上に改善することができる.このSAS-SD技術は難水溶性成分の水溶性を高度に改善するが, ガラス転移温度 (Tg) が低く保存安定性に乏しいことが分かっていた.また他の固体分散技術と同様に, 難水溶性成分の溶解時間にも改善の余地が大きかった.そこでいくつかのモデル薬剤と糖を用いてSAS-SDを作成し,そのSAS-SD試料を難水溶性成分の融点以上の温度で熱処理(アニーリング)すると難水溶性成分の対水溶解性, Tgともに改善することが分かった.この難水溶性物質の溶解挙動の変化機構について基礎的な知見を得るためDSC等により固体分散試料の熱特性, 相互作用状態を評価した.その結果,アニーリングすることで糖分子間の相互作用密度が緩和され, 糖分子がより安定な構造をとっているためであることが示唆された.さらに,同じ骨格構造は持つ複数の誘導体(ibuprofen等)をモデル薬剤としてSAS-SD試料を作成し,アニーリング条件と薬剤の対水溶解性,およびモデル薬剤の物理化学的特性の関係について検討した結果,SAS-SDおよびそのアニーリングによる薬剤の対水溶解性の改善度は薬剤の融点を用いて相関できることを見出した.さらにSAS-SDの調製手法・条件の検討を行った.具体的にはSAS-SD試料を真空foam乾燥と噴霧乾燥の二種類の方法で調製し,その物性を比較・検討した.

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  • ニードル刺激によるfoam dryingの乾燥効率の高度化および乾燥履歴の画一化

    研究課題/領域番号:15K14206  2015年04月 - 2017年03月

    日本学術振興会  科学研究費助成事業  挑戦的萌芽研究

    今村 維克

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    配分額:3770000円 ( 直接経費:2900000円 、 間接経費:870000円 )

    減圧下で溶液の発泡(foaming)させるfoam dryingは熱安定性の低い成分を含む溶液に対して有効であるが,発泡が生じる乾燥段階の制御が困難なことが多く,一定の乾燥履歴を得ることは難しい.これに対し,液体試料をある程度脱溶媒した上でneedleを刺し抜きすると減圧により直ちにfoamingを生じることを見出した.本研究では,このneedle刺激による溶液の乾燥履歴を画一化する技術を確立することをめざし,needle刺激条件や溶質-溶媒の組み合わせが,真空乾燥における発泡確率に及ぼす影響について調査した.また,needle刺激がfoamingを誘発するメカニズムについても検討した.

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  • 糖類アモルファスマトリクスに収着した水分の状態および機能の非単一性

    研究課題/領域番号:23560908  2011年 - 2013年

    日本学術振興会  科学研究費助成事業  基盤研究(C)

    今村 維克

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    配分額:5070000円 ( 直接経費:3900000円 、 間接経費:1170000円 )

    糖によって形成された非晶質固体(アモルファスマトリクス)は食品・医薬品において不可欠な素材である.本研究では,種々の糖類アモルファス試料について,個々の状態にある収着水の量を解析し,各試料の物理化学的特性と突き合わせることで,個々の収着状態にある水の物理化学的特性に対する寄与(機能)を明らかにした.約20種類の糖類を用いてアモルファス試料を作成し,それらの水分収着特性(水分収着等温線)とガラス転移温度(Tg),および収着状態を解析した.

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  • 酵素の精密制御固定化技術の確立と次世代高機能化バイオリアクターの構築

    研究課題/領域番号:22560771  2010年 - 2012年

    日本学術振興会  科学研究費助成事業  基盤研究(C)

    中西 一弘, 今村 維克, 今中 洋行

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    配分額:4550000円 ( 直接経費:3500000円 、 間接経費:1050000円 )

    本研究では、酵素・タンパク質を多孔性担体に固定化する際に問題となる固体表面との接触あるいは化学結合に際して生じる酵素・タンパク質の変性・失活と活性発現に不利な配向を防ぐ方法の開発を目的として研究を行った。具体的には、酵素をシリカやポリスチレンなどの固体表面に特異的親和性を有するペプチドタグを連結することにより、酵素を固体表面にほぼ不可逆的に固定化できるだけではなく、付着状態の構造と配向の制御が可能であることを示唆する結果を得た。

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  • 糖類アモルファスマトリクスのゼロモビリティー温度に基づく不安定物質の超高度安定化

    研究課題/領域番号:20560702  2008年 - 2010年

    日本学術振興会  科学研究費助成事業  基盤研究(C)

    今村 維克

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    配分額:4420000円 ( 直接経費:3400000円 、 間接経費:1020000円 )

    糖によって形成されたアモルファスマトリクスは,不安定物質を包括安定化する働きがある.一方,申請者は糖分子の赤外線吸収(IR)スペクトルの温度依存性からTgより数十℃低いある温度(T*)に糖分子間水素結合が高度に維持される温度域が存在することを見出した.このT*以下の温度領域では,糖分子のモビリティーが極端に制限され,マトリクス内に包括した酵素の失活がT*を境として顕著に大きくなることがわかった.さらに,種々の糖類からなるアモルファスマトリクスのT*を測定するとともにT*を上昇させる物質の探索を行った

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  • 親水性ポリスチレン高親和性ペプチドを用いた革新的蛋白質相互作用解析システムの創製

    研究課題/領域番号:19656221  2007年 - 2008年

    日本学術振興会  科学研究費助成事業  萌芽研究

    中西 一弘, 今村 維克, 今中 洋行

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    配分額:3300000円 ( 直接経費:3300000円 )

    本年度は、親水性ポリスチレン親和性ペプチドを用いた、1)コアストレプトアビジン(Core-SA)-DNA相互作用解析と2)相互作用に関与する機能性ペプチド部位を連結したクッションタンパク質を用いた相互作用解析を計画したが、両課題共に、初期の予定を達成できた。1)の課題においては、4量体のCore-SAに連結する親和性ペプチド(PSペプチド)の個数を変えて、その固定化特性とオリゴDNAとの相互作用をELISA法で検出した。結果として、4つのPSペプチドを連結したCore-SAは共存する高濃度牛血清アルブミンとの競合に影響されることなく、最も強固に固定化できることを示した。さらに、ビオチン標識オリゴDNAを連結したペプチドタグ連結コアストレプトアビジンは、市販のSA-coated plateよりも高い検出感度を示すことがわかった。一方、2)の課題に関しては,システイン合成酵素を構成するO-acetylserine sulfhydryrase-A(OASS-A)とそのC末端で相互作用することが知られているserine acetyltransferase(SAT)との間の相互作用に着目した。SATのC末端20残基からなるオリゴペプチドを、そのN末端にPSペプチドを連結したクッションタンパク質(RNaseHII)のC末端に連結し、オリゴペプチドのC末端アミノ酸を変化させてOASS-Aとの相互作用をELISA法で検出した結果、C末端アミノ酸がイソロイシンの野生型ペプチドを提示させた場合にのみ、相互作用することがわかった。同様に、Strep-tagIIをクッションタンパク質に提示したが、そのN末端をクッションタンパク質に向けて結合した場合にのみstreo-tactinと強い相互作用を示した。これらの結果から、本研究で検討した、親水性PS特異的親和性ペプチを用いる相互作用解析方法は、極めて高い検出感度と特異性を有することが明らかにされた。

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  • 糖類アモルファスマトリクスのゼロモビリティー温度と不安定物質の半永久的保存

    研究課題/領域番号:18760594  2006年 - 2007年

    日本学術振興会  科学研究費助成事業  若手研究(B)

    今村 維克

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    配分額:3500000円 ( 直接経費:3500000円 )

    糖によって形されたアモルファスマトリクスは,タンパク質などの不安定物質を包括すると,その劣化を抑制する働きがある.この糖類アモルファスマトリクスによる包括安定化作用は,マトリクスを構成する糖分子の状態と密接な関係があると考えられる.従って,室温以下の低温域においては,極めて高度な保存性が期待できる.本研究では,昇温過程における糖分子の赤外線吸収(IR)スペクトルを連続的に測定できる温度操作フーリエ変換赤外分光分析(TS-FTIR)により,-20℃からTg前後の温度域における糖分子間相互作用(水素結合)の形成状態を解析した.すなわち,糖水酸基の伸縮振動に起因するIR吸収帯のピーク波数を温度に対してプロットした.その結果,ピーク波数は温度上昇に伴い高波数側にシフトしていくが,Tgより数十℃低いある温度(T*)においてもピーク波数の増加勾配が顕著に小さくなることが分かった.これより,糖分子間水素結合が極めて安定に存在する温度域(<T*)が存在することが示唆された.また,その温度域の上限(T*)はsucroseで顕著に低く,糖の分子量とともに高くなる傾向が見られた.
    一方,糖類アモルファスマトリクスの構造エンタルピーの緩和過程を示差走査熱量分析(DSC)により測定し,異なる温度における糖分子のモビリティーを評価・比較した.その結果,上述のT*以下の温度領域では,各糖試料の構造エンタルピーの緩和が認められなかった.さらに,酵素を用いて糖類アモルファスマトリクスで包括したときの失活速度の温度依存性を調べたところ,T*以下では酵素の失活がほとんど生じないことが分かった.以上のことから,T*は糖分子のモビリティーが極端に制限される温度領域の上限値を表しており,この温度以下では極めて高度な包括安定化作用が発現することが明らかとなった.

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  • 製造プロセスで発生する付着および脱離の分子機構と洗浄操作の高効率化

    研究課題/領域番号:16206076  2004年 - 2007年

    日本学術振興会  科学研究費助成事業  基盤研究(A)

    中西 一弘, 今村 維克, 今中 洋行

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    配分額:46800000円 ( 直接経費:36000000円 、 間接経費:10800000円 )

    各種製造プロセスにおいて、製品製造に使用された各種装置や配管などの洗浄は、製品の安全性及び装置性能の維持に必要不可欠な重要な単位操作である。特に、食品製造プロセスやバイオプロセスにおいては、タンパク質など多様な成分からなる"汚れ"が装置壁面に強く相互作用するので、汚れを除去する洗浄操作には、多量のエネルギーと洗剤及び時間が費やされている。本研究では,汚れのモデル成分としてタンパク質、ペプチドや様々な低分子化合物を取り上げ、その金属酸化物表面への付着機構に着目して実験的、理論的解析を行った。その結果、1)低分子化合物、ペプチド及びタンパク質の表面への可逆吸着と不可逆的吸着の分子機構、2)種々の物質の付着形態及び配向の解析、3)金属酸化物の等電点及び溶液pHの付着に及ぼす影響とその機構、4)タンパク質の付着に伴う構造変化などについて明らかにした。さらに、金属酸化物だけではなく、プラスチック表面やガラス表面へのタンパク質の付着形態及び付着に関与するペプチドのアミノ酸配列についても検討した。プラスチック及びガラス表面上でもタンパク質は構造変化を起こすこと、逆に構造変化を抑制し、その配向を制御するためには、適切な表面親和性ペプチドを用いる必要があることを示した。一方、金属酸化物表面からの汚れ成分の脱離に関しては、申請者らが開発した過酸化水素電気分解処理に着目して、その脱離速度と脱離機構及び共存塩類の影響について詳細に検討した。本研究の結果、タンパク質の付着機構を分子レベルで解明することにより、各種製造プロセスで発生するタンパク性汚れの軽減と洗浄効率の改善のみならず、様々なバイオテクノロジー分野における先端技術の開発が可能であることが明らかにされた。

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  • 高度なタンパク質安定化作用と物理的安定性を兼備した新規糖類アモルファス構造の創製

    研究課題/領域番号:15760558  2003年 - 2004年

    日本学術振興会  科学研究費助成事業  若手研究(B)

    今村 維克

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    配分額:3500000円 ( 直接経費:3500000円 )

    目的
    高度なタンパク質安定化作用と物理的安定性を併せ持つ新規な糖類アモルファスマトリクスの構築を最終目標として,高度なタンパク質安定化作用を有する少糖と高い物理的安定性を有する多糖を組み合わせた少糖-多糖複合アモルファスの物理特性およびマトリクス内における分子間相互作用について検討した.
    実験方法
    スクロ-スおよびデキストランを様々な割合(0〜100%)で含む水溶液を調製した.一部の試料溶液には塩を0.1〜100mMになるように添加した.試料溶液を液体窒素で急速凍結して室温で1日凍結乾燥した.その後,さらにP_2O_5を入れた真空デシケータ内(37℃)で3日間絶乾処理することにより,少糖-多糖複合アモルファス試料を得た.少糖-多糖複合アモルファス内の分子間相互作用を解析するため,試料を一定相対湿度(RH 11,23,33%)下で平衡化後,含水率,ガラス転移温度(Tg)を測定した.試料の含水率は重量法およびKarl-Fischer法により測定し,ガラス転移温度は示差走査熱分析により測定した.
    結果
    スクロースーデキストラン複合アモルファス試料の水分収着特性を解析した結果,複合試料の平衡含水率はスクロースおよびデキストランそれぞれ単体のときの値から算出される理論値よりも低くなることが分かった.これはスクロースとデキストランが共存することでスクロース-デキストラン分子間に相互作用が形成し,それぞれ単体で存在するときよりも水和サイトが減少したことを意味している.一方,スクロース-デキストラン複合アモルファス試料のガラス転移温度に及ぼす塩の影響を調べた結果,リン酸塩の共存によるガラス転移温度の変化は認められなかったが,塩化ナトリウムや塩化アンモニウムはガラス転移温度を低下させることが分かった.

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  • 食品製造プロセスで発生する多成分系汚れの構造解析と脱離機構の解明

    研究課題/領域番号:13450319  2001年 - 2003年

    日本学術振興会  科学研究費助成事業  基盤研究(B)

    中西 一弘, 今村 維克, 崎山 高明

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    配分額:15100000円 ( 直接経費:15100000円 )

    本研究では食品製造プロセスで発生する汚れの付着挙動と構造解析,脱離機構の解明及び固体表面との相互作用の解析を目的として,主たる汚れ成分であるタンパク質およびその構成成分であるペプチドの金属表面への付着機構および複合汚れとしてのタンパク質/タンニン(酸)複合汚れの構造及び付着機構の解明を行った。さらに、本研究中に考案した過酸化水素(H_2O_2)から水酸化ラジカルを発生させる新規な洗浄方法による汚れの脱離機構および洗浄速度に関して詳細な検討を行った。主な成果の概要を下記に記す。
    1.タンパク質と金属固体表面との間の相互作用を解析するために、1)金属表面に付着しているタンパク質をプロテアーゼで分解して生成するペプチドの組成から解析する方法、2)タンパク質分子内の付着に関与するペプチドの変異ペプチドを調製して、その付着挙動から解析する方法の2つのアプローチを行った。その結果、複数の酸性アミノ酸(Asp、Glu)が隣接するペプチド部位がタンパク質分子内の直接の付着部位であること、ペプチドについては、酸性アミノ酸の数が多いほど、付着力及び付着量共に増加することが明らかになった。ペプチドの吸着等温線は、不可逆的な付着量と可逆的な付着(Langmuir型)の2つのモードの和として表すことができた。
    2.金属平板を希薄な過酸化水素溶液に接触させ、-0.2〜-0.8V程度の負電位を印加すると、金属平板上で水酸化(OH)ラジカルが発生することを見い出した。本方法を、タンパク質で汚染された金属表面の洗浄に適用した結果、室温でも数分以内にほぼ完全に脱離することを見い出すとともに、脱離速度に及ぼす諸因子を解析した。
    3.複合汚れのモデルとして、タンニン-タンパク質(β-Lactoglobulin:β-Lg)およびリン酸-タンパク質複合汚れを取り上げ、これらの汚れのステンレス表面における形成機構について詳細に解析した。さらに、この複合汚れのアルカリ洗浄とH_2O_2-電気分解洗浄における脱離挙動を種々の条件下で調べた。高感度FT-IR(RAS-IR)により、残存している残存物質の種類及び量を解析した。PH、処理温度の違いによる、金属表面と直接相互作用している付着物質の種類及び脱離速度に及ぼす影響を詳細に調べた。

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  • 高度なタンパク質安定化機能を有する糖類アモルファスマトリクスの創製

    研究課題/領域番号:12750664  2000年 - 2001年

    日本学術振興会  科学研究費助成事業  奨励研究(A)

    今村 維克

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    配分額:2200000円 ( 直接経費:2200000円 )

    目的 不安定なタンパク質を安定化する手法の一つとして、タンパク質を糖などの安定化剤とともに凍結乾燥することでタンパク質を糖のアモルファスマトリクスに包埋し、保存中の変性、失活を低減する手法がある。このタンパク質安定化剤としてはスクロースなどの低分子量の糖は高い安定化作用を有するものの、吸湿や熱により容易にコラプスするという欠点がある.一方,デキストランなどの多糖類は安定化作用は極めて低いものの,物理的安定性に優れている。従って,高い安定化作用を有する少糖と高い物理的安定性を有する多糖を組み合わせることで,高度な安定化作用と物理的安定性を併せ持つ新規な糖類アモルファスマトリクスを構築出来る可能性がある.この新規な安定化剤である少糖-多糖複合アモルファスの物理的安定性(ガラス転移温度,Tg)およびタンパク質安定化作用について検討した。
    方法 少糖としてsucrose, erythritol,多糖として分子量の異なる2種類のdextran(MW1500, 6000),タンパク質としてlactate dehydrogenase (LDH)を用いた.これらの少糖・多糖を様々な割合(0〜100%)で含む凍結乾燥試料を作成し,凍結乾燥直後のLDHの相対残存活性,一定相対湿度下で平衡化したときのTgを測定した.
    結果 sucrose-dextran, erythritol-dextran試料のTgはいずれの相対湿度においてもdextran含有率が40%以上で顕著に上昇することが分かった。一方,凍結乾燥直後の残存酵素活性は,sucrose crythritol単体の場合はいずれも50%であったのに対し,sucrose-dextran試料では最大70%, erythrito1-dextran試料では最大95%となった.これらのことより少糖-多糖複合アモルファスマトリクスの極めて高い有効性が明らかとなった.

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  • 固体表面に対する配向の精密制御されたタンパク質の創製

    研究課題/領域番号:12019251  2000年

    日本学術振興会  科学研究費助成事業  特定領域研究(A)

    中西 一弘, 今村 維克, 崎山 高明

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    配分額:2000000円 ( 直接経費:2000000円 )

    タンパク質(酵素)の固体表面に対する付着は、分析、反応、生体材料設計など生物生体工学の多岐の分野においてみられれ現象であり、それぞれの応用を考える際には付着に対する親和性と配向の制御が重要な鍵となる。そこで、本研究では、タンパク質の固体表面に対する相互作用と直接付着に関与する部位の解明を行うと共に固体表面に対する親和性と配向の制御されたタンパク質の創製を最終目的としている。タンパク質として分子量が小さく、性質や立体構造が明らかにされているウシ膵臓ribonucleaseA(RNaseA)に着目して、固体表面に直接相互作用するタンパク質分子内ペプチド部位の解明と固体表面に対する親和性及び配向の制御された酵素の創製を行うために、変異RNaseAの調製法を確立すると共にその付着挙動を調べた。変異RNaseAとしては、ステンレス表面に対する付着部位と推定されている、^<47>His-Glu-Ser-Leu-Ala-Asp-Val^<57>中のAlaをAspに置換したAla55Asp変異RNaseAとGluとAspのそれぞれあるいは両方をAlaに置換した、Glu55Ala変異RNaseAとAsp55Ala変異RNaseA、及びPS Latexに対し強い付着を示す^<117>Pro-Val-His-Phe-Asp-Ala-Ser-Val^<124>中のPheをAlaに置換したPhe120Ala変異RNaseAを調製した。いずれの場合も変異RNaseA遺伝子をプラスミドpBXRに組み込んだ発現プラスミドを作成し、大腸菌(BL21)内で発現させた。培養大腸菌を回収後に超音波破砕を行い、封入体を回収した。回収した封入体を変性・再生後にイオン交換クロマトグラフィーとゲル濾過により精製した。精製酵素はSDS-PAGE上で均一であることを確認した、精製酵素の収率は10%前後であった。精製酵素のステンレス粒子及びPS Latexに対する付着特性を調べた。予備的検討の結果、予想した通りにAla55Asp変異RNaseAはステンレス粒子に対して野生型RNaseAよりも強い付着力を示したが、Phe120Ala変異RnaseAでは変化がみられなかった。

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  • 固体表面に対する親和性と配向の精密制御されたタンパク質の創製

    研究課題/領域番号:11132250  1999年

    日本学術振興会  科学研究費助成事業  特定領域研究(A)

    中西 一弘, 今村 維克

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    配分額:2000000円 ( 直接経費:2000000円 )

    本研究では、タンパク質と固体表面の直接の相互作用に着目して、固体表面に対する親和性と配向の制御されたタンパク質の創製を最終目的として、1)固体表面に直接相互作用するタンパク質分子内ペプチド部位を特定する方法の確立と、2)ペプチド部位の固体表面に対する親和性の強さとそのアミノ酸組成・長さ・二次構造等との関係を明らかにすることである。本年度はモデルタンパク質として、主として分子量が小さく三次元構造が明らかにされている、ウシ膵臓由来リボヌクレアーゼA(RNaseA)を、固体表面としてステンレス粒子とポリスチレンラテックス(Pslatex)を用いて、固体表面に直接相互作用する部位の同定と相互作用に関する知見を得ることを目的とした。RNaseAをカルボキシメチル化した後、トリプシン処理し、得られたペプチド混合液あるいは合成したペプチドを試験液に溶解し、吸着等温線を測定した。主な成果は次の3点に要約される。
    1)RNaseAの各pHにおけるステンレスに対する吸着等温線はいずれもLangmuir型を示したが、高pH領域では、殆ど吸着がみられなかった。Pslatexの場合では吸着等温線はpHに殆ど依存せず、不可逆吸着を示した。
    2)トリプシン処理によって得られたRNaseA由来ペプチドを用いて吸着実験を行ったところ、ステンレスに対しては酸性アミノ酸残基を含むペプチド10が、Pslatexに対しては、疎水性アミノ酸残基を多く含むペプチド9が顕著に吸着していた。3)ペプチド9-1の疎水性アミノ酸残基であるIleをGlyに、ペプチド9-3のPheをGlyに、10-1の酸性アミノ酸残基GluとAspをGlyに置換したペプチドを合成し、吸着特性の変化を調べたところ、いずれのペプチドも吸着力は激減した。これらの結果から、ステンレスに対する吸着には酸性アミノ酸残基が、Pslatexに対する吸着には、疎水性アミノ酸残基による疎水相互作用とベンゼン環同士のπ-π相互作用が深く関与していることが示唆された。

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  • 食品製造プロセスにおいて発生する汚れの装置表面への付着と脱離機構の解明

    研究課題/領域番号:10450287  1998年 - 2000年

    日本学術振興会  科学研究費助成事業  基盤研究(B)

    中西 一弘, 今村 維克, 崎山 高明

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    配分額:14100000円 ( 直接経費:14100000円 )

    本研究では、1)タンパク性汚れ、ペプチド、アミノ酸の付着特性の解明、2)タンパク質の固体表面に対する直接の付着部位の決定、及び3)酵素及び過酸化水素/紫外線により発生する水酸化ラジカルを利用した汚れの洗浄機構の解明の3点に焦点を絞り検討した。固体表面としてはステンレス表面を取り上げた。タンパク質としてβ-ラクトグロブリン(β-Lg)、牛血清アルブミン(BSA)、ゼラチン(Gel)を取り上げ、ステンレス表面に対する付着特性を種々の条件下で明らかにした。β-LgとBSAについては、中性より酸性側では、不可逆的な吸着挙動を示した。付着温度が上昇するに従い、β-LgとBSAの付着量は増加したが、これはシステイン残基間の結合反応やS-S交換反応による界面凝集反応により多分子層吸着が形成されるためであることが示唆された。一方、Gelに関しては温度上昇に伴い、付着量は低下した。さらに、ペプチド類とアミノ酸の付着挙動を調べたところ、酸性条件下では、酸性アミノ酸(GluとAsp)が付着に関与することが明かとなった。β-Lgのステンレス表面に対する直接の付着部位を同定するために、プロテアーゼ消化により調製されたペプチド断片の付着特性を明らかにした。その結果、アミノ酸配列の125番目から136番目のPeptide T5(Thr-Pro-Glu-Val-Asp-Asp-Glu-Ala-Leu-Glu-Lys)領域が有力な付着部位の一つであることが示唆された。このペプチドとステンレス平板間の相互作用をFT-IRを用いて調べたところ、酸性アミノ酸の側鎖カルボキシル基が解離した状態で付着していることが明らかにされた。さらに、タンパク質で汚染された表面の種々の条件下で酵素洗浄と水酸化ラジカル洗浄を行い、その挙動をアルカリ洗浄の結果と比較した。

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  • 糸状菌の潜在機能を活用できる新規な培養システムの開発とその特性解明

    研究課題/領域番号:10555291  1998年 - 2000年

    日本学術振興会  科学研究費助成事業  基盤研究(B)

    中西 一弘, 長澤 透, 今村 維克, 崎山 高明

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    配分額:13200000円 ( 直接経費:13200000円 )

    本研究では申請者らが開発した、原理的にカビの培養に適した膜面液体培養法(Membranae-Surface Liquid Culture;MSLC)を、1)Aspergillus oryzae NRRL484を用いるコージ酸の生産、2)遺伝子組換えAspergillus oryzae AGL142-72を用いるα-グルコシダーゼの生産、及び3)Monascus pilosus kによる紅色色素の生産に適用し、培養特性を振盪フラスコ培養法や寒天プレート培養法の結果と比較検討した。1)については、いずれの培養条件においても、MSLCでは振盪フラスコ培養法に比較して、生成物コージ酸の生成速度、最大コージ酸収率のみならず、菌体当たりの生成量も高い値を示した。2)については、検討した全ての培養条件下においてMSLCでは振盪フラスコ培養法に比較して、酵素生成量と菌体量当たりの酵素生成量共に高い値を示した。窒素源として無機窒素(硝酸ナトリウム)を用いた場合、MSLCで得られる菌体量は振盪フラスコ培養の結果よりも著しく高いという興味深い現象を見い出した。振盪フラスコ培養法に比較してMSLCによる高い酵素生産性の理由を、酸素供給、酵素の分泌性、及びmRNAレベルの3つの観点から考察した。酸素供給については振盪フラスコ培養において、酸素容量係数を種々変化させて検討を行ったが、菌体量と酵素生成量共に殆ど変化しなかった。MSLCで培養された菌体のmRNA発現量と酵素分泌性は振盪フラスコ培養法の結果に比較して著しく高いことがわかった。Monascus pilosus kによる紅色色素の生産においても、色素生成量はMSLCの方が高い値を示した。MSLCで高い物質生産性が得られた理由として、いずれの場合も酸素供給の影響は殆どなく、菌糸の足場が存在する表面上で静置培養することが主な理由であることが示された。

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  • 糖-タンパク質間水素結合のタンパク質の安定化に対する寄与とその評価

    研究課題/領域番号:10750548  1998年 - 1999年

    日本学術振興会  科学研究費助成事業  奨励研究(A)

    今村 維克

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    配分額:2000000円 ( 直接経費:2000000円 )

    タンパク質水溶液に糖を添加し凍結乾燥すると,タンパク質は糖によって形成されたアモルファス構造に包埋され,その結果,タンパク質の安定性が顕著に向上する.この糖によるタンパク質安定化機構についてはこれまで多くの研究がなされているが,依然未解明な点が多く,実際の安定化操作の改善に繋がる知見は乏しい.そこで本研究では糖としてスクロース,マルトース,トレハロース,マルトトリオース,および平均分子量1500,6000のデキストラン,タンパク質として牛血清アルブミン(BSA)を様々な割合で含有する糖-タンパク質凍結乾燥資料を作成し,試料中におけるタンパク質の構造保持の度合いを比較した.タンパク質の構造保持度はフーリエ変換赤外分光分析計(FT-IR)を用いて二次構造を解析することにより評価し,各種糖アモルファスの物理的特性は示差走査熱量計(DSC)を用いて分析した.また,タンパク質安定化作用との密接な関係が注目されている糖-タンパク質間水素結合を水分収着特性から定量し,タンパク質の構造保持度との関係について検討を加えた.その結果,タンパク質の構造保持度をαへリックスの含有率を指標として評価した場合,構造保持の度合いはスクロース>マルトース>トレハロース>マルトトリオース>デキストラン1500>デキストラン6000の順となり,スクロースを用いた場合,糖添加量が1g/g-BSAのとき水溶液中の値とほぼ一致した.各種糖アモルファスのガラス転移温度(Tg)を測定した結果,Tgが低い糖ほどタンパク質の構造保持度が高いことが分かった.これはTgが低い糖アモルファスほど糖分子の構造柔軟性が高く,タンパク質との相互作用が形成され易いためと考えられる.また,糖-タンパク質間水素結合の形成度を評価した結果,糖-タンパク質間水素結合の形成度は構造保持の度合いと大略傾向は一致することが明らかになった.

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  • 固体表面に対する親和性と配向の精密制御された抗体の創製

    研究課題/領域番号:10145237  1998年

    日本学術振興会  科学研究費助成事業  特定領域研究(A)

    中西 一弘, 今村 維克

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    配分額:2000000円 ( 直接経費:2000000円 )

    平成10年度は、比較的分子量の小さいRNAseやβ-ラクトグロブリン(β-Lg)をモデルタンパク質として用いて、固体表面に直接接触しているぺプチド部位の特定と相互作用の強さに関して検討を試みた。直接付着に関与するぺプチド部位を特定する方法としては、タンパク質のプロテアーゼ消化により得られるぺプチド類の付着挙動から推定する間接的な方法を用いた。タンパク濃度一定条件下では、等電点近傍のpH5近傍で最大の吸着量を示した。吸着等温線(30℃)は見掛けLangmuir型を示した。酸性領域(pH3〜4)では、イオン強度が高いほど、吸着平衡定数は大きい結果となった。β-Lgをトリプシンで処理することにより14種類のペプチドを単離した。各ペプチドの組成と配列を一次構造とアミノ酸分析により同定した。ペプチド混合溶液あるいは各ペプチド単独の溶液を用いて種々の条件下(PH、温度、イオン強度)で吸着挙動を調べた。β-Lgが高い吸着を示した酸性pH領域では酸性アミノ酸を複数個含むペプチドがステンレス粒子に対する親和性が高いことが判明した。これは、ペプチド分子中の酸性アミノ酸残基がマイナスに誘起されて、ステンレス表面のプラスの電荷と多点結合するためであることが示唆された。同様にトリプシン処理によって得られたカルポキシメチル化RNaseA由来ペプチドを単離(10種類)し、そのステンレス粒子及びラテックスに対する吸着挙動をペプチド混合溶液を用いて検討した。いずれのペプチドもステンレスに対する吸着は、殆どみられなかった。一方、ラテックスに対する吸着は疎水性アミノ酸残基が多く含むペプチドにおいて強いことから、ラテックスに対する吸着には疎水性相互作用が寄与していることが示唆されたが、ペプチドとRNaseAの吸着挙動には、β-Lgのような明確な相関関係はみられなかった。

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  • バイオおよび食品製造プロセスにおける洗浄操作の高効率化によるエミッションの削減

    研究課題/領域番号:10141228  1998年

    日本学術振興会  科学研究費助成事業  特定領域研究(A)

    中西 一弘, 今村 維克

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    配分額:2000000円 ( 直接経費:2000000円 )

    本研究の目的は,バイオプロセスにおいて膨大なエミッションを生み出す洗浄操作の合理化ある.平成10年度はOHラジカルの高い酸化力に着目し,(1)固体表面汚れの効率的除去と(2)洗浄排水の低減の可能性を検討した.
    (1) 付着汚れの除去への応用は次のような原理に基づいている.H_2O_2水溶液を汚れの付着した固体表面に薄い液膜として流し,その上部から紫外線を照射すると,H_2O_2の紫外線分解によりOHラジカルが発生し,付着状態および脱着・遊離した汚れ物質を酸化分解する.さらに,光照射により汚れ物質の脱着が促進される.モデルは汚れ物質として脂肪酸を付着させたステンレス表面を用いて,付着汚れのラジカル酸化除去効果および紫外線による脱離促進効果を検証した.脂肪酸の付着・残留量はRAS-FT-IRにより定量した.その詰果,水流だけでは脂肪酸はほとんど脱離しないが,紫外線の照射により顕著に脂肪酸が脱離することが分かった.
    (2) 洗浄排水処理へのラジカル酸化分解の応用については,モデル有機物質として色素を含む着色水溶液のUV-H_2O_2処理における脱色特性を明らかにした.いづれの色素についても脱色反応は一次反応に従い,見かけの一次反応速度定(K)のH_2O_2濃度依存性を求めたところ,H_2O_2濃度に最大値が存在した.これはH_2O_2濃度の増加伴い,OHラジカルの発生速度とともにUVの減衰が顕著になるためであると考えられる.そこで,紫外線照射下のH_2O_2-色素水溶液におけるラジカル反応(H_2O_2の紫外線分解反応,OHラジカルとH_2O_2との反広,OHラジカルと色素との反応)とH_2O_2および色素による紫外線の減衰を考慮したモデルを設定し,kの理論値を計算した.その結果,実験値と良好に一致し,本モデルにより有機物質のラジカル酸化速度に対する条件の最適化が可能である.

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  • ナノポーラス担体への固定化による酵素の有機溶媒中での安定性と反応特性の改変

    研究課題/領域番号:09555256  1997年 - 1998年

    日本学術振興会  科学研究費助成事業  基盤研究(B)

    今村 維克, 崎山 高明, 宇多川 隆, 中西 一弘, 今村 維克

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    配分額:1100000円 ( 直接経費:1100000円 )

    本研究では、水溶液中では困難な反応を可能にせしめる有機溶媒中での酵素反応を実用化するために問題となる基礎を確立することを目的とした。サーモライシン(EC3.4.24.4)により触媒されるアスパルテーム前駆体Z-L-Asp-L-PheOMe(Z-;N-(Benzyloxycarbonyl),-OMe;Methyl ester)合成反応(縮合反応)を取り上げ、酵素をサブミクロンオーダーの細孔径を有する種々の多孔性担体に固定化した。固定化酵素の有機溶媒中での安定性に及ぼす諸因子の把握とその機構の解明を行った。固定化酵素の安定性に関しては、特に水難溶性有機溶媒中での失活速度について検討を行い、1)固定化酵素の失活は主として担体内部での自己消化によるものであること、2)自己消化の程度は有機溶媒の種類、水分濃度、担体の種類に影響されるが、これは固定化されている酵素分子周囲の水相の状態が異なるためであることを明らかにした。一方、合成速度を定量的に推定するために、水/有機溶媒二相系での分配挙動及び有機溶媒飽和の水溶液中での合成初速度に関して解析を行った。基質の酸成分と塩基成分間のイオンペア形成を考慮した分配モデルと有機溶媒飽和緩衝液中での合成初速度から、固定化酵素の有機溶媒中でのジペプチド合成初速度の推定を行ったが、酢酸エチル中では酵素の有効利用度の低下を考慮することにより精度良く推定できることを示した。

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  • Adsorption & desorption of protein, peptide, & amino acids

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    資金種別:競争的資金

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  • タンパク質・ペプチド・アミノ酸の金属壁面への付着・脱離

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    資金種別:競争的資金

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  • ラジカル酸化反応の排水処理,固体表面の洗浄への応用

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    資金種別:競争的資金

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  • Application of radical oxidation to waste water treatment & cleaning of Solid Surfaces

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    資金種別:競争的資金

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  • Stabilization of protein by sugars

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    資金種別:競争的資金

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  • Adsorption & desorption of protein, peptide, & aminoocids

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    資金種別:競争的資金

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  • Application of radical oxidation to waste water treatment & cleaning of Sclid Surfaces

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    資金種別:競争的資金

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  • Stabilization of protein by sugar

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    資金種別:競争的資金

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  • 糖によるタンパク質の安定化

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    資金種別:競争的資金

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  • 実践応用化学 (2022年度) 前期  - その他

  • 応用化学ゼミナール1 (2022年度) 前期  - その他

  • 応用化学ゼミナール2 (2022年度) 後期  - その他

  • 応用化学基礎 (2022年度) 後期  - 月3~4,火3~4

  • 応用化学特別研究 (2022年度) 通年  - その他

  • 材料プロセス各論3 (2022年度) 夏季集中  - その他

  • 材料機能化学特論7 (2022年度) 夏季集中  - その他1~8

  • 混合と反応 (2022年度) 第1学期  - 月4,木2

  • 熱力学Ⅲ (2022年度) 第1学期  - 月3,木1

  • 物理化学及び演習2 (2022年度) 第1学期  - 月3~4,木1~2

  • 物理化学2 (2022年度) 第1学期  - 月3~4,木1~2

  • 物理化学2 (2022年度) 第1学期  - 月3~4,木1~2

  • 生物化学工学 (2022年度) 前期  - 木1,木2

  • バイオプロセス工学演習 (2021年度) 通年  - その他

  • バイオ分子間相互作用解析学 (2021年度) 前期  - その他

  • 化学・生命系入門 (2021年度) 第1学期  - 金1~2

  • 化学工学1A (2021年度) 第3学期  - 水1,水2

  • 化学工学1B (2021年度) 第4学期  - 水1,水2

  • 化学工学2A (2021年度) 第1学期  - 火1,火2

  • 化学工学2B (2021年度) 第2学期  - 火1,火2

  • 化学工学1 (2021年度) 3・4学期  - 水1,水2

  • 化学工学2 (2021年度) 1・2学期  - 火1,火2

  • 実践応用化学 (2021年度) 前期  - その他

  • 応用化学ゼミナール1 (2021年度) 前期  - その他

  • 応用化学ゼミナール2 (2021年度) 後期  - その他

  • 応用化学基礎 (2021年度) 後期  - 月3~4,火3~4

  • 応用化学特別研究 (2021年度) 通年  - その他

  • 材料プロセス各論3 (2021年度) 夏季集中  - その他

  • 材料機能化学特論3 (2021年度) 集中  - その他

  • 物理化学及び演習2 (2021年度) 第3学期  - 月3,月4,木3,木4

  • 物理化学2 (2021年度) 第3学期  - 月3,月4,木3,木4

  • 生物化学工学 (2021年度) 前期  - 木1,木2

  • バイオプロセス工学演習 (2020年度) 通年  - その他

  • バイオ分子間相互作用解析学 (2020年度) 後期  - その他

  • 化学工学1A (2020年度) 第3学期  - 水1,水2

  • 化学工学1B (2020年度) 第4学期  - 水1,水2

  • 化学工学2A (2020年度) 第1学期  - 火1,火2

  • 化学工学2B (2020年度) 第2学期  - 火1,火2

  • 化学工学1 (2020年度) 3・4学期  - 水1,水2

  • 化学工学2 (2020年度) 1・2学期  - 火1,火2

  • 実践応用化学 (2020年度) 前期  - その他

  • 応用化学ゼミナール1 (2020年度) 前期  - その他

  • 応用化学ゼミナール2 (2020年度) 後期  - その他

  • 応用化学基礎 (2020年度) 後期  - 月3~4,火3~4

  • 応用化学特別研究 (2020年度) 通年  - その他

  • 材料プロセス各論3 (2020年度) 集中  - その他

  • 材料プロセス各論2 (2020年度) 夏季集中  - その他

  • 材料機能化学特論7 (2020年度) 冬季集中  - その他

  • 物理化学及び演習2 (2020年度) 第3学期  - 月3,月4,木3,木4

  • 物理化学2 (2020年度) 第3学期  - 月3,月4,木3,木4

  • 生物化学工学 (2020年度) 前期  - 木1,木2

▼全件表示