Cloning and Expression in Escherichia coli of the Glutamate Recemase Gene from Pediococcus pentosaceus(Biological Chemistry)
スポンサーリンク
概要
- 論文の詳細を見る
The glutamate racemase (EC 5.1.1.3) gene of a lactic acid bacterium, Pediococcus pentosaceus, was cloned into Escherichia coli C600 with a vector plasmid, pBR322. The requirement of L-glutamate for the growth of E. coli in the minimum medium containing D-glutamate and the formation of a red pigment in a coupled enzyme reaction mixture were used to select clones expressing glutamate racemase activity. Glutamate racemase overproduced as 0.3〜2.0% of the total soluble proteins in a clone carrying the plasmid pICR221, 10.3 kb of DNA, was purified from cell extracts about 130-fold to homogeneity. The purified enzyme has a molecular weight of about 40,000 and is a single polypeptide chain. Glutamate is the sole substrate for the enzyme. Unlike many other amino acid racemases, glutamate racemase is devoid of cofactors: there is no evidence for pyridoxal 5'-phosphate or FAD in the ultraviolet spectrum of the purified enzyme, and the enzyme is not inactivated by carbonyl reagents such as hydroxylamine and sodium borohydride.
- 社団法人日本農芸化学会の論文
- 1986-11-23
著者
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TANAKA Hidehiko
Laboratory of Microbiology, Institute for Chemical Research, Kyoto University
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SODA Kenji
Laboratory of Microbiology, Institute for Chemical Research, Kyoto University
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Soda Kenji
Laboratory Of Microbial Biochemistry Institute For Chemical Research Kyoto University
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Soda Kenji
Institute For Chemical Resarch Kyoto University
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Nakajima Nobuyoshi
Laboratory Of Microbial Biochemistry Institute For Chemical Research Kyoto University:(present Offic
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TANIZAWA Katsuyuki
Laboratory of Microbial Biochemistry, Institute for Chemical Research, Kyoto University
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谷澤 克行
大阪大学
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Soda K
Department Of Biotechnology Kansai University
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Tanaka Hidehiko
Laboratory Of Biochemistry Kyoto College Of Pharmacy
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Tanizawa Katsuyuki
Laboratory Of Microbial Biochemistry Institute For Chemical Research Kyoto University
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