2021/12/27 更新

写真a

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

学位

  • 工学博士 ( 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   Graduate School, Division of Engineering  

    - 1997年

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  • 京都大学    

    - 1997年

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    国名: 日本国

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  • 京都大学   Faculty of Engineering  

    - 1992年

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  • 京都大学   工学部   化学工学科

    - 1992年

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    国名: 日本国

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経歴

  • 岡山大学   大学院自然科学研究科   教授

    2012年6月 - 現在

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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

  • 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

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

    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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    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.2021.127744

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

    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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    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.2021.127174

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

    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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    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.2021.08.002

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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

    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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    掲載種別:研究論文(学術雑誌)   出版者・発行元:Informa UK Limited  

    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: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

    Takumi Deguchi, Takato Nakahara, Koreyoshi Imamura, Naoyuki Ishida

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

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    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

    1 - 11   2020年9月

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    担当区分:最終著者   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1080/07373937.2020.1822863

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

    Shuichiro Kimura, Koreyoshi Imamura, Junichiro Futami

    Biotechnology Progress   36 ( 5 )   2020年9月

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    掲載種別:研究論文(学術雑誌)   出版者・発行元:Wiley  

    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: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: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 招待 査読

    145 ( 17 )   5801 - 5807   2020年

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    掲載種別:研究論文(学術雑誌)  

    DOI: 10.1039/d0an00998a

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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:責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

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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

    108 ( 6 )   2056 - 2062   2019年6月

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

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

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

    Colloids and Surfaces B: Biointerfaces   166   262 - 268   2018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:Elsevier B.V.  

    The effect of the properties of a protein on its adsorption to a metal surface in the presence of external electric potential was investigated. Protein adsorption processes at different surface potentials were measured for fifteen types of proteins using an in-situ ellipsometry. The tested proteins were classified into three groups, based on the amount of protein that was adsorbed as a function of the surface potential: In First group of proteins, an increasing trend for the amount adsorbed with a more positive surface potential was found
    The amount adsorbed of α-chymotrypsinogen A and ribonuclease A (Second group) were roughly constant and independent of the applied surface electric potentials
    In Third group, the amount adsorbed decreased with increasing surface potential. This protein classification was correlated with the isoelectric points of the proteins (First group: ≤9.3
    Second group: 9.3–10
    Third group: &gt
    10). Increasing the pH positively and negatively shifted the surface potentials, allowing ß-lactoglobulin (First group) and lysozyme (Third) to become adsorbed, respectively. The surface potential range for protein adsorption was also markedly shifted depending on the metal substrate type. These findings were interpreted based on the electrostatic interactions among the protein, surface hydroxyl groups, and the applied external electric field.

    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

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

    COLLOIDS AND SURFACES B-BIOINTERFACES   159   118 - 124   2017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  

    Enzymatic cleaning is a potentially useful method for removing proteinaceous fouling from solid surfaces under mild conditions. Herein, the influence of an external electric field on the enzymatic cleaning of a metal surface fouled with a protein was investigated. The model fouling protein (BSA; lysozyme) was prepared on a stainless steel (St) surface, and the resulting surface subjected to enzymatic cleaning with an electric potential being applied to the St plate. Trypsin, alpha-chymotrypsin, and thermolysin were used as model proteases. The amounts of protein remaining on the plate before and during the cleaning process were measured by means of a reflection absorption technique using Fourier transform infrared spectroscopy. In the case for BSA fouling, the cleaning efficiency of the protease tended to increase at more negative applied potentials. Whereas, there was an optimum applied potential for removing the lysozyme fouling. Atomic force microscopy analyses indicated that applying an adequate range of electric potential enhanced the enzymatic removal of protein fouling inside scratches on the St plate surface. These findings suggest the existence of two modes of electrostatic interactions for the external electric field, one with protease molecules and the other with digested fragments of the fouling protein. (C) 2017 Elsevier B.V. All rights reserved.

    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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    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.

    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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    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.

    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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    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.

    DOI: 10.1016/j.colsurfb.2016.07.042

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

    Hiroyuki Imanaka, Koki Dare, Koreyoshi Imamura

    NEW BIOTECHNOLOGY   33   S72 - S72   2016年7月

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    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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    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 >= 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.

    DOI: 10.1016/j.foodchem.2015.11.097

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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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    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

    DOI: 10.1002/btpr.2232

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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.

    DOI: 10.1016/j.apt.2015.10.017

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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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    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.

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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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    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.

    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 <= 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 <= 2.8 x 10(-4) g-lipid (g-water)(-1). The size and the lipid amount continued to increase gradually at the lipid amount >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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    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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  • 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 >= 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.

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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)  

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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 >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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  • 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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  • 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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  • 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(-) -> 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.

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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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  • 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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  • 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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  • 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H<SUB>2</SUB>O<SUB>2</SUB>-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.H<SUB>2</SUB>O<SUB>2</SUB>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, H<SUB>2</SUB>O<SUB>2</SUB>-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, H<SUB>2</SUB>O<SUB>2</SUB>-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, H<SUB>2</SUB>O<SUB>2</SUB>-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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  • 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.

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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.

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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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  • 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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  • 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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  • 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.

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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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  • 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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  • 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 <I>Streptomyces mobaraensis</I> 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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  • 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.

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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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  • 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.

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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.

    DOI: 10.1252/jcej.30.609

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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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MISC

  • 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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    記述言語:日本語  

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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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    記述言語:日本語  

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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-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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    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/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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    その他リンク: http://search.jamas.or.jp/link/ui/2013125672

  • 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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  • 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 )   2009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  

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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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  • 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.

    DOI: 10.1271/bbb.70052

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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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  • 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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  • 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.

    DOI: 10.1016/j.jcis.2004.06.081

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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.

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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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  • 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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  • 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.

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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.

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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.

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  • ステンレス表面に吸着したタンパク質のH<sub>2</sub>O<sub>2</sub>-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

  • 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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  • 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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  • 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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  • 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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  • 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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  • 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.

    DOI: 10.1271/bbb.65.865

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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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  • UV/H<sub>2</sub>O<sub>2</sub>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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  • 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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  • 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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  • 紫外線/H<sub>2</sub>O<sub>2</sub>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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  • 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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  • 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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  • 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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  • 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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  • 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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受賞

  • 化学工学会奨励賞

    2002年  

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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

  • 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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  • 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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  • 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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  • 糖によるタンパク質の安定化

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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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  • 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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