Cloning, Sequence Analysis, and Expression in Escherichia coli of the Gene Encoding Monovalent Cation-activated Levodione Reductase from Corynebacterium aquaticum M-13
スポンサーリンク
概要
- 論文の詳細を見る
The gene encoding (6R)-2,2,6-trimethyl-1,4-cyclohexanedione (levodione) reductase was cloned from the genomic DNA of the soil isolate bacterium Corynebacterium aquaticum M-13. The gene contained an open reading frame consisting of 801 nucleotides corresponding to 267 amino acid residues. The deduced amino acid sequence showed approximately 35% identity with other short chain alcohol dehydrogenase/reductase (SDR) superfamlly enzymes. The probable NADH-binding site and three catalytic residues (Ser-Tyr-Lys) were conserved. The enzyme was sufficiently produced in recombinant Escherichia coli cells using an expression vector pKK223-3,and purified to homogeneity by two-column chromatography steps. The enzyme purified from E.coli catalyzed stereo-and regio-selective reduction of levodione, and was strongly activated by monovalent cations, such as K^+, Na^+, and NH_4^+, as was the case of that from C.aquaticum M-13. To our knowledge, this is the first sequencing report of a monovalent cation-activated SDR enzyme.
- 社団法人日本農芸化学会の論文
- 2001-04-23
著者
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Shimizu Sakayu
Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University
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和田 政裕
Faculty Of Pharmaceutical Sciences Josai University
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TAKAGI Hiroshi
Department of Surgery II, University School of Medicine
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HIRAI TAKANORI
Komaki Municipal Hospital
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Shimazu S
Div. Appl. Life Sci. Grad. Sch. Agric. Kyoto Univ.
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Shimizu Sakayu
Division Of Applied Life Science Graduate School Of Agriculture Kyoto University
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WADA Masaru
Department of Bioscience, Fukui Prefectural University
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NAKAMORI Shigeru
Department of Bioscience, Fukui Prefectural University
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YOSHISUMI Ayumi
Department of Bioscience, Fukui Prefectural University
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Wada M
Osaka Univ. Osaka
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Wada Masaru
Department Of Biology College Of Liberal Arts And Sciences Tokyo Medical And Dental University
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Wada M
Tokyo Medical And Dental Univ. Chiba Jpn
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Takagi H
Department Of Food Science And Technology Faculty Of Agriculture Nagoya University:(present Address)
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Takagi Hiroshi
Second Department Of Surgery Nagoya University
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Yoshisumi Ayumi
Department Of Bioscience Fukui Prefectural University
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Nakamori Shigeru
Dept. Biosci. Fukui Pref. Univ.
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Nakamori Shigeru
Department Of Bioscience Faculty Of Bioscience Fukui Prefectural University
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Wada Masaru
Department Of Bioscience Fukui Prefectural University
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Takagi H
Department Of Bioscience Fukui Prefectural University
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Takagi Hiroshi
Department Of Medicine And Molecular Science Gunma University Graduate School Of Medicine
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Nakamori S
Department Of Bioscience Fukui Prefectural University
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Takagi H
Graduate School Of Biological Sciences Nara Inst. Of Sci. And Technol.
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Wada M
Division Of Applied Bioscience Research Faculty Of Agriculture Hokkaido University
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Takagi Hiroshi
Department Of Bioscience Fukui Prefectural University
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Wada Masaru
Department Of Biology College Of Liberal Arts And Sciences Tokyo Medical And Dental Univ.
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Wada Masaru
Department Of Biology And Chemistry College Of Liberal Arts And Sciences Tokyo Medical Dental Univer
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Takagi Hiroshi
Department Of Bioscience Faculty Of Bioscience Fukui Prefectural University
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