Comparative Characterization of New Glutaryl 7-ACA and Cephalosporin C Acylases
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概要
- 論文の詳細を見る
We performed a comparative characterization of three new cephalosporin acylases which were prepared from E. coli recombinant strains and found originally from Pseudomonas sp. A14,Bacillus laterosporus J1 and Pseudomonas diminuta N176. Both A14 and N176 acylases consisted of two non-identical subunits (α, β) whose molecular weights were 28,000 (α), 61,000 (β) and 26,000 (α), 58,000 (β), respectively, whereas J1 acylase consisted of a single peptide with molecular weight of 70,000. The maximum specific activities of A14,J1 and N176 acylases for glutaryl 7-ACA were 7.1,5.3 and 100 units/mg, respectively, and that of N176 acylase for cephalosporin C was 3.1 units/mg. The K_m values of glutaryl 7-ACA for A14,J1 and N176 acylases were 2.1,3.2 and 2.6 mM, respectively, and that of cephalosporn C for N176 acylase was 4.8 mM. A14,J1 and N176 acylases exhibited differential activities for cephalosporins having an aliphatic dicarboxylic acid in the acyl side chain and only N176 acylase showed an activity for cephalosporin C. N176 acylase as well as A14 acylase also showed a weak activity for a cephalosporn derivative having a heterocyclic carboxylic acid in the side chain. A14,J1 and N176 acylases catalyzed the reverse reaction to synthesize glutaryl 7-ACA from 7-ACA and glutaric acid, although the rate of the synthesis was 10 to 10^5 fold slower than that of hydrolysis. The activities of the cephalosporin acylases were considerably inhibited by the reaction products, 7-ACA and glutaric acid. The types of the inhibition by 7-ACA and glutaric acid were both competitive. A14,J1 and N176 acylases were thermostable, their residual activities exceeding more than 90% after treatment at 50℃ for 1 h at their optimal pHs.
- 社団法人日本生物工学会の論文
- 1992-03-25
著者
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Ono Hisayo
Department of Biotechnology, Graduate School of Engineering, Osaka University
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Ono Hisayo
Department Of Biotechnology Graduate School Of Engineering Osaka University
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Ono Hisayo
Department Of Fermentation Technology Osaka University
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Aramori Ichiro
Pharmacological Research Laboratories Fujisawa Pharmaceutical Co. Ltd.
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Aramori Ichiro
Exploratory Research Laboratories Fujisawa Pharmaceutical Co. Ltd.
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Ishii Y
Pharmacological Research Laboratories And Exploratory Research Laboratories
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KOJO Hitoshi
Advanced Technology Platform Research Laboratory Fujisawa Pharmaceutical Co., Ltd.
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FUKAGAWA Masao
Advanced Technology Platform Research Laboratory Fujisawa Pharmaceutical Co., Ltd.
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FUKAGAWA MASAO
Exploratory Research Laboratories
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TSUMURA MANA
Exploratory Research Laboratories
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IWAMI MORITA
Exploratory Research Laboratories
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ONO HIROKI
Manufacturing Technology Laboratories
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ISHITANI YOSUKE
Manufacturing Technology Laboratories
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KOJO HITOSHI
Exploratory Reseach Laboratories
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KOHSAKA MASANOBU
Research and Development Group
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UEDA YOSHIO
Research and Development Group
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IMANAKA HIROSHI
Research and Development Group
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Ishii Y
Fermentation Development Laboratories Fujisawa Pharmaceutical Co. Ltd.
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Kojo H
Advanced Technology Platform Research Laboratory Fujisawa Pharmaceutical Co. Ltd.
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Fukagawa Masao
Advanced Technology Platform Research Laboratory Fujisawa Pharmaceutical Co. Ltd.
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Ono Hisayo
Department Of Biotechnology Faculty Of Engineering Osaka University
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Ono H
Manufacturing Technology Laboratories:fujisawa Pharmaceutical Co. Ltd.
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KOHSAKA MASANOBU
Pharmacological Research Laboratories and Exploratory Research Laboratories
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Ueda Yoshio
New Drug Research Laboratories Fujisawa Pharmaceutical Co. Ltd.
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Iwami Morita
Exploratory Research Laboratories:fujisawa Pharmaceutical Co. Ltd.
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Tsumura Masanori
Exploratory Research Laboratories:fujisawa Pharmaceutical Co. Ltd.
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