MISC - IMAMURA Koreyoshi

パッキングのコントロールにより,難結晶性蛋白質の結晶化を促進する結晶化法

小坂恵, 山田秀徳, 二見淳一郎, 多田宏子, 今村維克, 玉田太郎

日本細胞生物学会大会(Web)   71st   2019

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パッキング相互作用に着目したタンパク質結晶化法

小坂恵, 山田秀徳, 二見淳一郎, 多田宏子, 今村維克, 玉田太郎

日本結晶学会年会講演要旨集   2019   2019

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難結晶性蛋白質のパッキングをコントロールする結晶化法の開発と評価

小坂恵, 山田秀徳, 二見淳一郎, 多田宏子, 今村維克, 玉田太郎

日本結晶学会年会講演要旨集   2018   2018

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タンパク質のパッキングをコントロールする疎水性残基導入

小坂恵, 山田秀徳, 二見淳一郎, 多田宏子, 今村維克, 玉田太郎

日本結晶学会年会講演要旨集   2017   107   2017.11

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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

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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   2016.9

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Surfactant-Free Solid Dispersion of Fat-Soluble Flavor in an Amorphous Sugar Matrix, Based on Over-Dissolution of Sugar in Organic Media

( 28 )   67 - 76   2015

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2P-064 Screening of a novel Pd affinity peptide tag and characterization of tag conjugated protein immobilization

Matoba Haruka, Ishida Naoyuki, Imamura Koreyoshi, Imanaka Hiroyuki

67   190 - 190   2015

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Other Link： http://search.jamas.or.jp/link/ui/2016231956

3P-195 Influences of sequence at the 5'-terminal region and media components on Escherichia coli recombinant expression system

Abo Hirotaka, Hara Hirofumi, Umetsu Mitsuo, Ishida Naoyuki, Imamura Koreyoshi, Imanaka Hiroyuki

67   319 - 319   2015

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Other Link： http://search.jamas.or.jp/link/ui/2016232284

標的遺伝子の5’末端周辺配列が大腸菌発現系に及ぼす影響

安保紘高, 原啓文, 石田尚之, 今村維克, 今中洋行

日本農芸化学会大会講演要旨集(Web)   2015   2015

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2P-020 Influence of codon bias and media components on the Escherichia coli recombinant protein expression system and transcriptional profile of E. coli

Abo Hirotaka, Miki Syunya, Hara Hirofumi, Yamazoe Atsushi, Hosoyama Akira, Tsuchikane Keiko, Imamura Koreyoshi, Imanaka Hiroyuki

66   111 - 111   2014

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Other Link： 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   2013.5

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2P-076 The influence of orientation and density of immobilized ligand molecule on biomolecular interaction

Matsushita Runa, Ishida Naoyuki, Imamura Koreyoshi, Imanaka Hiroyuki

65   123 - 123   2013

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Other Link： http://search.jamas.or.jp/link/ui/2014247513

3P-180 Screening of novel gold affinity peptide and its application on protein immobilization

Shigemori Yojiro, Ishida Naoyuki, Imamura Koreyoshi, Takahashi Yuichiro, Imanaka Hiroyuki

65   233 - 233   2013

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4Cp02 Screening of NFkB(p50) affinity peptides by adopting functional biomoleculer immobilization

Takimoto Takayuki, Miyahara Tetsuya, Imamura Koreyosi, Imanaka Hiroyuki, Kondou Eisaku, Nakanishi Kazuhiro

64   195 - 195   2012

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Other Link： http://search.jamas.or.jp/link/ui/2013125671

4Cp03 Examination of the site specific peptide drug screening method transcription factor, FOXP3, as the target

Kawasaki Kazuki, Matsumoto Kohei, Imanaka Hiroyuki, Imamura Koreyoshi, Kondou Eisaku

64   195 - 195   2012

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Other Link： http://search.jamas.or.jp/link/ui/2013125672

11p04 Investigation of overexpression of ε-Lysine acylase from Streptomyces mobaraensis in Escherichia coli as a host

Imanaka Hiroyuki, Taguchi Yuzo, Uchida Tatsuya, Kumon Kazuki, Kuroki Kentaro, Imamura Koreyoshi, Nakanishi Kazuhiro

63   58 - 58   2011

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2Ep03 Screening of NFkB(p50)inhibitory peptides using affinity tag mediated functional protein immobilization

Takimoto Takayuki, Miyahara Tetsuya, Imanaka Hiroyuki, Imamura Koreyoshi, Kondo Eisaku, Nakanishi Kazuhiro

63   145 - 145   2011

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

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

JOURNAL OF BIOSCIENCE AND BIOENGINEERING   110 ( 3 )   281 - 287   2010.9

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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.

DOI： 10.1016/j.jbiosc.2010.03.010

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Temperature-Scanning Fourier Transform IR Spectroscopic Analysis of Amorphous Sugar Matrices : Changes in Interaction States with Temperature

IMAMURA Koreyoshi

Calorimetry and thermal analysis.   37 ( 4 )   154 - 161   2010.8

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

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

Memoirs of the Faculty of Engineering, Okayama University   44   73 - 83   2010.1

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2P-1103 Influence of amino acid sequence on its translocation to E. coli cell surface by autotransporter

IMANAKA Hiroyuki, UEDA Hisako, IMAMURA Koreyoshi, NAKANISHI Kazuhiro

22   34 - 34   2010

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

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

22   71 - 71   2010

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3P-1085 Identification of a key amino acid residue for the formation of cysteine synthase complex using a highly functional peptide immobilization

SHIMIZU Yuki, IMANAKA Hiroyuki, IMAMURA Koreyoshi, NAKANISHI Kazuhiro

22   79 - 79   2010

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

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

Journal of bioscience and bioengineering   108 ( 1 )   S161   2009.11

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Efficient Ethanol Production from Wheat Bran by Enzymatic Saccharification Using Commercially Available Enzyme Products and Fermentation Using Bakers' Yeast

KOREISHI Mayuko, IMANAKA Hiroyuki, IMAMURA Koreyoshi, KARIYAMA Masahiro, NAKANISHI Kazuhiro

Seibutsu-kougaku Kaishi   87 ( 5 )   216 - 223   2009.5

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We aimed to produce ethanol efficiently by enzymatic saccharification coupled with ethanol fermentation using wheat bran as a biomass. For this purpose, we examined pretreatment conditions of biomass and a combination of commercially available enzyme products for saccharification. When a 3.3% (w/v) wheat bran suspension was pretreated using subcritical water at 140℃ with a holding time of 0h, followed by saccharification at 45℃ using a mixture of commercially available Meiselase (composed of cellulase, cellobiase/β-glucosidase, and xylanase) and Novozyme^[○!R] 188 (composed of cellobiase/β-glucosidase, α-amylase, and glucoamylase) at a ratio of 4 to 1 by weight, glucose was released at 10.6g/l, which corresponded to 90% of the value calculated on the basis of the total amount of glucose-based saccharides contained in the wheat bran. On the other hand, from a pretreated 33% (w/v) wheat bran suspension, 85g/l glucose was produced, which corresponded to only 72% of the theoretical value, owing to the inhibitory effect of glucose on β-glucosidase activity. However, when fermentation was carried out at 30℃ following 24-h saccharification of the pretreated wheat bran using a mixture of Meiselase and Novozyme^[○!R] 188, the inhibitory effect of glucose was considered to be reduced and as a result, after 21-h fermentation, ethanol was produced at 5.2% (w/v), which corresponded to 88% of the theoretical maximum value.

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2Ea05 Investigation of peptide surface display system by autotransporter

UEDA Hisako, YAMASHITA Mai, IMANAKA Hiroyuki, IMAMURA Koreyoshi, NAKANISHI Kazuhiro

21   59 - 59   2009

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2Ip05 Investigation of Peptide-Protein Interaction Detection System by Cushion Protein

IMANAKA Hiroyuki, KUNIKATA Toshinobu, YAMANOUCHI Mai, IMAMURA Koreyoshi, NAKANISHI Kazuhiro

21   126 - 126   2009

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2Da01 Construction of expression systems of a novel aminoacylase from Streptomyces mobaraensis

OOI Manami, NAKATANI Yasuyuki, KOREISHI Mayuko, IMANAKA Hiroyuki, IMAMURA Koreyoshi, NAKANISHI Kazuhiro

21   44 - 44   2009

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3Ga07 Development of rapid and sensitive detection system of human FOXP3-DNA interaction

MAEKAWA Masami, IMANAKA Hiroyuki, IMAMURA Koreyoshi, KONDO Eisaku, NAKANISHI Kazuhiro

20   186 - 186   2008

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Other Link： http://search.jamas.or.jp/link/ui/2009066377

2Aa03 Purification and Characterization of Recombinant Monoacylglycerol Lipase from Rhodanobacter sp. strain LP7315

TERAKAWA Shuji, MORI Masashi, IMANAKA Hiroyuki, IMAMURA Koreyoshi, NAKANISHI Kazuhiro

20   59 - 59   2008

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3Da04 Immobilization of Biomolecules by Cushion Protein on Solid Surfaces

IMANAKA Hiroyuki, YAMAZUMI Daisuke, YANAGITA Keisuke, IMAMURA Koreyoshi, NAKANISHI Kazuhiro

20   126 - 126   2008

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Enzymatic modification of proteolytic fragments of food proteins with fattyacid using a novel acylase

Annual report of study for food under the supporting program by The Skylark Food Science Institute   ( 21 )   109 - 115   2008

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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

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

BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY   71 ( 6 )   1582 - 1586   2007.6

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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.

DOI： 10.1271/bbb.70052

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1C11-2 Enzymatic synthesis of functional compounds by the acyl-transfer reaction catalyzed by penicillin V acylase from Streptomyces mobaraensis

KOREISHI Mayuko, ISE Yuuichi, TANI Kazuha, IMANAKA Hiroyuki, IMAMURA Koreyoshi, NAKANISHI Kazuhiro

19   79 - 79   2007

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2F14-4 Production of protease and gene expression analysis of Aspergillus oryzae cultivated by membrane-surface liquid culture and shaking flask culture

TANAKA Soukichi, FENG Bin, IMANAKA Hiroyuki, IMAMURA Koreyoshi, NAKANISHI Kazuhiro

19   141 - 141   2007

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Other Link： http://search.jamas.or.jp/link/ui/2008290979

1C11-1 Characterization and cloning of long-chain aminoacylase and shortchain aminoacylase from Streptomyces mobaraensis

NAKATANI Yasuyuki, KOREISHI Mayuko, IMANAKA Hiroyuki, IMAMURA Koreyoshi, NAKANISHI Kazuhiro

19   79 - 79   2007

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1C09-2 Influence of signal peptide on the construction of cell surface display system by autotransporter

TATSUMOTO Wataru, IMANAKA Hiroyuki, IMAMURA Koreyoshi, NAKANISHI Kazuhiro

19   77 - 77   2007

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ポリスチレン親和性ペプチド標識タンパク質を用いる迅速免疫測定

中西一弘, 熊田陽一, 今中洋行, 今村維克

日本食品工学会年次大会講演要旨集   8th   2007

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Stabilization of Proteins during Freeze-drying and Storage by "Sugar Glass"

IMAMURA Koreyoshi

81 ( 949 )   908 - 912   2006.11

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

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

JOURNAL OF BIOSCIENCE AND BIOENGINEERING   101 ( 5 )   434 - 439   2006.5

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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.

DOI： 10.1263/jbb.101.434

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ポリスチレン表面に親和性を示すペプチドタグを付加した酵素の設計と特性解析

今石大輔, 熊田陽一, 今中洋行, 今村維克, 崎山高明, 中西一弘

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

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ポリスチレン親和性ペプチド標識タンパク質を用いる迅速免疫測定

熊田陽一, 今中洋行, 今村維克, 加藤滋雄, 中西一弘

化学工学会年会研究発表講演要旨集   71st   2006

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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’05)   2005

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2E11-4 Orientated immobilization of protein by use of affinity peptide specific to polystyrene

KUMADA Yoichi, IMANAKA Hiroyuki, IMAMURA Koreyoshi, KATOH Shigeo, SAKIYAMA Takaaki, NAKANISHI Kazuhiro

17   153 - 153   2005

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Sugar-Protein Interaction and Stabilization of Protein in Amorphous Sugar Matrix(Papers presented at the Seminar, "Amorphous Science for Food and Pharmaceutical System")

IMAMURA Koreyoshi

Cryobiology and cryotechnology   51 ( 1 )   31 - 35   2005

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

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ポリスチレン親和性ペプチド融合タンパク質の吸着特性とその応用

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

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

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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   2004.11

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

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

JOURNAL OF BIOSCIENCE AND BIOENGINEERING   98 ( 3 )   200 - 206   2004.9

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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： 10.1263/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   2004.6

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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   2004.5

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

DOI： 10.1263/jbb.97.322

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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   2004.1

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

DOI： 10.1016/j.colsurfb.2003.08.010

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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

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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

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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

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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

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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   2003.11

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

DOI： 10.1016/S0021-9797(03)00700-8

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

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

JOURNAL OF COLLOID AND INTERFACE SCIENCE   265 ( 1 )   49 - 55   2003.9

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

DOI： 10.1016/S0021-9797(03)00329-1

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放線菌由来新規アシラーゼによるカプサイシン誘導体の合成とその抗酸化性の評価

KOREISHI MAYUKO, CHO TOKUMIN, KAWANAMI KAZUKO, IMAMURA YUKIKATSU, SAKIYAMA TAKAAKI, MATSUMURA YASUO, NAKANISHI KAZUHIRO

日本食品工学会年次大会講演要旨集   4th   117   2003.7

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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   2003.6

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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   2003.2

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

DOI： 10.1002/jps.10305

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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   2003.2

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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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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

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ペプチドのステンレス表面への吸着に対する酸性アミノ酸残基の寄与

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

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

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

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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

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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   2002.10

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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.

DOI： 10.1021/la025893i

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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   2002.10

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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.

DOI： 10.1021/la025893i

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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   2002.10

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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.

DOI： 10.1002/jps.10218

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A Novel Cleaning Method Using Electrolysis of Hydrogen Peroxide

IMAMURA Koreyoshi, NAKANISHI Kazuhiro

77 ( 899 )   800 - 805   2002.9

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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

MIZUTANI YUKIKO, MATSUMURA YASUO, IMAMURA TADAKATSU, NAKANISHI KAZUHIRO, MORI TOMOHIKO

日本食品科学工学会大会講演集   49th   152   2002.8

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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   2002.6

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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(-) --> .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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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   2002.6

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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(-) --> .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   2002.1

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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   2002.1

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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.

DOI： 10.1006/jcis.2001.7967

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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   2002.1

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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.

DOI： 10.1006/jcis.2001.7967

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

Koreyoshi Imamura, Atsushi Fukushima, Keisuke Sakaura, Takuo Sugita, Takaharu Sakiyama, Kazuhiro Nakanishi

Journal of Pharmaceutical Sciences   91 ( 10 )   2175 - 2181   2002

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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, Tg, increased to a considerable extent when the dextran content was equal to or higher than 50%, while the increase in Tg at dextran contents lower than 50% was small. A marked increase in Tg 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. © 2002 Wiley-Liss, Inc.

DOI： 10.1002/jps.10218

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Influences of the Suspension Method of Thermolysin in Organic Solvents on Enzyme Activity

Keizo Tanaka, Koreyoshi Imamura, Takaharu SAkiyama, Kazuhiro Nakanishi

Japan Journal of Food Engineering   3 ( 1 )   15 - 23   2002

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Thermolysin (EC 3. 4. 24. 4) was suspended in organic solvents containing a small amount of water (aqueous buffer solution) by two different methods, and the catalytic activity as well as the conformational changes in the enzyme were compared. In one method (Method 1), dried powder enzyme was directly suspended in organic solvents containing a small amount of water while in the second method (Method 2), the dried enzyme was first dissolved in the buffer solution and then added to the anhydrous organic solvents containing the same final water content as that used in Method 1. In Method 2, the enzymes showed a catalytic activity 2-10 times as high as that in Method 1. The ratio of the activity obtained in Method 2 to that in Method 1 strongly depended on kinds of organic solvents, and its dependency was reasonably explained by the ET(30) values, which represent the extent of polarity of an organic solvent. The Fourier transform infrared spectroscopy (FT-IR) analyses indicated that the enzyme conformation in Method 2 was closer to its native one than that in Method 1. The influences of the suspension method on the enzyme activities of the different forms of the enzyme (as donated, dialyzed, and cross-linked) and the courses of the enzyme activities during incubation in Methods 1 and 2 were also investigated. © 2002, Japan Society for Food Engineering. All rights reserved.

DOI： 10.11301/jsfe2000.3.15

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S219 Prospects for micro-bioreactor systems with respect to reactor and biocatalyst designs

Nakanishi Kazuhiro, Sakiyama Takaharu, Imamura Koreyoshi

14   21 - 21   2002

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308 Analysis and control of adsorption behavior of enzyme onto a solid surface

Ohnishi Kousuke, Imamura Koreyoshi, Sakiyama Takaharu, Nakanishi Kazuhiro

14   28 - 28   2002

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306 Water sorption and glass transition of freeze-dried disaccharidepolymer mixtures

Fukushima Atsushi, Sakaura Keisuke, Sugita Takuo, Imamura Koreyoshi, Sakiyama Takaharu, Nakanishi kazuhiro

14   28 - 28   2002

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310 Removal of protein adsorbed on a metal surface by H_2O_<2^->electrolysis treatment

Imamura Koreyoshi, Watanabe Ippei, Sakiyama Takaharu, Nakanishi Kazuhiro

14   29 - 29   2002

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309 Basic study on the control of enzyme binging onto polystyrene surfaces

Ueno Shunsuke, Sakiyama Takaharu, Imamura Koreyoshi, Nakanishi Kazuhiro

14   29 - 29   2002

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549 Various substrate specificities of a novel acylase from Streptomyces sp.

Koreishi Mayuko, Zhang Demin, Takayanagi Eiko, Hata Kazuki, Imamura Koreyoshi, Sakiyama Takaharu, Nakanishi Kazuhiro

14   85 - 85   2002

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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   2001.12

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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.

DOI： 10.1002/jps.1146

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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   2001.7

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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   2001.4

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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   2001.3

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

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

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

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UV/H₂O₂処理を利用したステンレス表面に付着したタンパク質の洗浄

34 ( 7 )   869 - 877   2001

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Enzymatic modification of β-lactoglobulin with N-fatty-acyl-dipeptides by transglutaminase from Streptomyces mobaraense.

Wada Eiko, Masuda Haruki, Imamura Koreyoshi, Sakiyama Takaharu, Adachi Shuji, Matsuno Ryuichi, Nakanishi Kazuhiro

13   158 - 158   2001

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Purification of capsaicin-hydrolyzing enzyme from Streptomyces sp.

Koreishi Mayuko, Zhang Demin, Imamura Koreyoshi, Sakiyama Takaharu, Adachi Syuji, Matsuno Ryuichi, Nakanishi Kazuhiro

13   159 - 159   2001

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pH-Responsive swelling of chitosan/chondroitin sulfate complex gels and its application.

Sakiyama Takaharu, Ikuta Yuji, Imamura Koreyoshi, Nakanishi Kazuhiro

13   282 - 282   2001

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Cleaning of a stainless steel surface fouled with protein by H_2O_2-electrolysis technique

Imamura Koreyoshi, Tada Yoichiro, Tanaka Hirofumi, Sakiyama Takaharu, Nakanishi Kazuhiro, Tanaka Atsumi, Yamada Yosuke

13   282 - 282   2001

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Cultivation characteristics of rerecombinant Aspergillus oryzae for production of enzyme

Shimamura Hiroko, Morita Masakazu, Yonashiro Ryota, Imammura Koreyoshi, Sakiyama Takaharu, Ninetoki Toshitaka, Nakanishi Kazuhiro

13   199 - 199   2001

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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   2000.11

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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   2000.9

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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.

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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   2000.9

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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.

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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   2000.9

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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.

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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   2000.8

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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.

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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   2000.7

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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   2000.7

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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   2000.4

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A novel advanced oxidation system using a combined UV/H2O2 technique was constructed for application to wastewater treatment. In this system, wastewater containing H2O2 flows along a channel with a flat surface in a thin film, onto which surface UV rays with a wavelength of 253.7 nm are irradiated. As a model wastewater, we used various dye solutions and investigated decoloration rates under various conditions differing in H2O2 concentration, temperature, UV illuminance, and so on. Based on the model, which takes into consideration the formation as well as the disappearance rate of hydroxyl radical (. OH), distribution of UV illuminance along the depth of the flow, and the rate of reaction between dye and . OH, we could simulate the course of dye decomposition. Furthermore, we showed the existence of an optimum H2O2 concentration for decomposition on the basis of this model.

DOI： 10.1252/jcej.33.253

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膜面液体培養法を用いた海洋植物プランクトンOscillatoria属の培養と特性解明

中西 一弘, 崎山 高明, 今村 維克

財団法人ソルト・サイエンス研究財団助成研究報告集   1998 ( 1 )   279 - 286   2000.3

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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

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紫外線/H₂O₂処理を利用した新規促進酸化システムの開発と色素水溶液の分解過程の速度論の解析

33 ( 2 )   253 - 261   2000

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

Mino Koshiki, Imamura Koreyoshi, Sakiyama Takaharu, Eisaki Naoki, Matsuyama Asahi, Nakanishi Kazuhiro, RITE

12   225 - 225   2000

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Synthesis of capsaicin analogues by aminoacylase.

Koreishi Mayuko, Kamakura Kaoru, Imamura Koreyoshi, Sakiyama Takaharu, Adachi Syuji, Matsuno Ryuichi, Nakanishi Kazuhiro

12   218 - 218   2000

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Enzymatic synthesis of N-acylamino acids in cosolvents.

Wada Eiko, Handa Masato, Imamura Koreyoshi, Sakiyama Takaharu, Adachi Shuji, Matsuno Ryuichi, Nakanishi Kazuhiro

12   219 - 219   2000

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Synthesis of nonproteinous amino acid by cysteine synthetase with acetyl-CoA regeneration system.

Ohno Katsuhiro, Mino Koshiki, Imamura Koreyoshi, Sakiyama Takaharu, Eisaki Naoki, Matsuyama Asahi, Nakanishi Kazuhiro

12   218 - 218   2000

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Maintenance of protein structure in dry state by amorphous sugar

Imamura Koreyoshi, Ogawa Tomohiro, Sakiyama Takaharu, Nakanishi Kazuhiro

12   280 - 280   2000

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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   1999.12

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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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Adsorption characteristics of tryptic fragments of bovine beta-lactoglobulin on a stainless steel surface

T Sakiyama, K Tanino, M Urakawa, K Imamura, T Takahashi, T Nagai, K Nakanishi

JOURNAL OF BIOSCIENCE AND BIOENGINEERING   88 ( 5 )   536 - 541   1999.11

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As a strategy for the analysis of the mode of protein adsorption onto stainless steel surfaces, peptides obtained by tryptic digestion of bovine beta-lactoglobulin were subjected to adsorption experiments after identification of their primary structures. In the presence of I 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 mM HNO3 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 beta-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   1999.4

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Various factors affecting the stability of thermolysin immobilized by cross-linking with glutaraldehyde were elucidated, particularly in the water-immiscible organic solvents such as ethyl acetate and tert-amyl alcohol. The main reason for enzyme inactivation in water-immiscible organic solvents was found to be autolysis in the water phase, which may surround the enzyme immobilized inside the support. By contrast, in water-miscible organic solvents thermal denaturation was the predominant cause of enzyme inactivation. Courses of inactivation were expressed by second-order kinetics in the initial stage, after which inactivation proceeded at a slower rate. The extent of autolysis was found to strongly depend on the kind of organic solvent, the water content, and type of support and these dependencies were explained by the difference in the amount and state of water inside the support. Thermolysin was immobilized onto Amberlite XAD-7 as a compact aggregate inside the support which may increase the stability of the enzyme. Finally, it was shown that the stability of the immobilized enzyme could be correlated with the logP value for water-miscible organic solvents and with the solubility of water for water- immiscible organic solvents.

DOI： 10.1016/S1389-1723(99)80095-4

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食品製造・バイオプロセスにおける洗浄とタンパク性汚れの付着(<特集>殺菌・洗浄のバイオテクノロジー)

中西 一弘, 今村 維克, 崎山 高明

生物工学会誌 : seibutsu-kogaku kaishi   77 ( 6 )   241 - 245   1999

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Characteristics and kinetics of cysteine synthetase from Escherichia coli.

Mino Koshiki, Imamura Koreyoshi, Sakiyama Takaharu, Eisaki Naoki, Matsuyama Asahi, Nakanishi Kazuhiro, RITE

11   170 - 170   1999

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Adsorption behaviors of protein, peptide, and amino acid onto stainless steel surface

Imamura Koreyoshi, Sato Masako, Tomura Junji, Okamoto Makoto, Sakiyama Takaharu, Nakanishi Kazuhiro, Takahashi Tokio, Nagai Tadashi

11   333 - 333   1999

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Production of pigment from Monascus pilosus k using membrane surface liquid culture

Tsushima Youko, Mino Akira, Imamura Koreyoshi, Sakiyama Takaharu, Nakanishi Kazuhiro, Suga Sadaharu, Danou Yasuhumi

11   220 - 220   1999

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Purification and characterization of serine acetyltransferases truncated at C-terminal regions.

Hiraoka Kenji, Mino Koshiki, Imamura Koreyoshi, Sakiyama Takaharu, Eisaki Naoki, Matsuyama Asahi, Nakanishi Kazuhiro, RITE

11   167 - 167   1999

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Synthesis of β-pyrazole alanine using cysteine synthetase coupled with acetyl-CoA regeneration system.

Ohno Katsuhiro, Mino Koshiki, Yamanoue Tsuyoshi, Imamura Koreyoshi, Sakiyama Takaharu, Eisaki Naoki, Matsuyama Asahi, Nakanishi Kazuhiro, RITE

11   184 - 184   1999

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Characteristics of enzyme production from recombinant Aspergillus oryzae using membrane surface liquid culture

Shimamura Hiroko, Ishida Natsuko, Sugita Takuo, Imamura Koreyoshi, Sakiyama Takaharu, Nakanishi Kazuhiro, Ozeki Kenji

11   220 - 220   1999

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Characteristics of production of kojic acid from Aspergillus oryzae NRRL 484 using membrane surface liquid culture

Sugita Takuo, Segawa Takayuki, Shimamura Hiroko, Imamura Koreyoshi, Sakiyama Takaharu, Nakanishi Kazuhiro

11   219 - 219   1999

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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   1998.6

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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.

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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   1998.4

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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.

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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

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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

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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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Influence of protein on phase transition of amorphous sugar

A. Koreyoshiimamur, Tetsuo Suzuki, Shinya Kirii, Takayuki Tatsumichi, Morio Okazaki

Journal of Chemical Engineering of Japan   31 ( 3 )   325 - 329   1998

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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 frceze-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. Copyright ©1998 The Society of Chemical Engineers,.

DOI： 10.1252/jcej.31.325

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新規に開発したカビの膜面液体培養法を用いた食品加工用酵素の生産に関する基礎研究

中西 一弘, 崎山 高明, 今村 維克

食に関する助成研究調査報告書   ( 11 )   51 - 58   1998

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Language：Japanese   Publisher：すかいら-くフ-ドサイエンス研究所  

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Adsorption of amino acids and poptides onto the surface of stainless steel particles.

Mimura Tomoya, Tanino Kei, Imamura Koreyoshi, Sakiyama Takaharu, Nakanishi Kazuhiro

9   190 - 190   1997

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Cultivation of mold by membrane-surface liquid culture and its scale-up

Segawa Takayuki, Imamura Koreyoshi, Sakiyama Takaharu, Nakanishi Kazuhiro

9   96 - 96   1997

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