An Approach to High-Level Production of Cephalosporin Acylase from Pseudomonas strain N176 in Escherichia coli
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概要
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The cultivation conditions for N176 cephalosporin acylase production were optimized using Escherichia coli JM109/pCK305S, which encodes a highly expressed mutant, C305S, in which Cys^<305>is altered to Ser, under the control of the trp promoter. Processing of the precursor protein to a two-chain active form in vivo was a key factor for high-level production of the enzyme. The active acylase was produced in a high amount under the following conditions : (i)addition of Trp in the second seed medium, (ii)the use of glycerol as a carbon source, (iii)induction with IAA at 9h, and(iv)a shift down in the cultivation temperature from 25 to 22.5℃ after induction. Under these high-level production conditions, the OD at 600 nm was found to increase in proportion to the number of colony forming units(CFUs), indicating that the CFU is an important index for the expression of the active enzyme.
- 1998-06-25
著者
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Yamashita Mitsuo
Department Of Biotechnology Graduate School Of Engineering Osaka University:department Of Applied Ch
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Yamashita M
Department Of Biotechnology Graduate School Of Engineering Osaka University
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NOGUCHI YUJI
Pharmacological Research Laboratories, Fujisawa Pharmaceutical Co. Ltd.
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YAMADA HISASHI
Pharmacological Research Laboratories, Fujisawa Pharmaceutical Co. Ltd.
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YAMAGUCHI MICHITAKA
Pharmacological Research Laboratories, Fujisawa Pharmaceutical Co. Ltd.
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WATANABE KATSUYUKI
Pharmacological Research Laboratories, Fujisawa Pharmaceutical Co. Ltd.
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FUJIMURA TAKAO
Pharmacological Research Laboratories, Fujisawa Pharmaceutical Co. Ltd.
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ISHII YOSHINORI
Pharmacological Research Laboratories, Fujisawa Pharmaceutical Co. Ltd.
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SAITO YOSHIMASA
Pharmacological Research Laboratories, Fujisawa Pharmaceutical Co. Ltd.
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Ishii Y
Pharmacological Research Laboratories And Exploratory Research Laboratories
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Noguchi Yuji
Fermentation Development Laboratories Fujisawa Pharmaceutical Co. Ltd.
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Fujimura Takao
Pharmacological Research Laboratories Fujisawa Pharmaceutical Co. Ltd.
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Ishii Yoshinori
Pharmacological Research Laboratories Fujisawa Pharmaceutical Co. Ltd.
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Ishii Y
Fermentation Development Laboratories Fujisawa Pharmaceutical Co. Ltd.
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Yamada Hisashi
Fermentation Development Laboratories Fermentation Development Laboratories Fujisawa Pharmaceutical
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Watanabe Katsuyuki
Pharmacological Research Laboratories Fujisawa Pharmaceutical Co. Ltd.
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Watanabe K
Division Of Applied Bioscience Graduate School Of Agriculture Hokkaido University
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Yamada H
Fermentation Development Laboratories Fermentation Development Laboratories Fujisawa Pharmaceutical
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Ishii Y
Ishii Orthopaedic And Rehabilitation Clinic
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Watanabe K
Marine Biotechnol. Inst. Iwate Jpn
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Yamashita Manabu
Research Institute For Bioresources Okayama University
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Fujimoto Tamotsu
Pharmacological Research Laboratories Fujisawa Pharmaceutical Co. Ltd.
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Matsumoto Yukimasa
Department Of Biotechnology Graduate School Of Engineering Osaka University
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