Cloning, Expression and Nucleotide Sequence of the L-2, 3-Butanediol Dehydrogenase Gene from Brevibacterium saccharolyticum C-1012

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概要

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• A 3-kbp DNA fragment including the L-2, 3-butanediol dehydrogenase(L-BDH)gene(budC)from the chromosomal DNA of Brevibacterium saccharolyticum C-1012 was cloned in Escherichia coli JM109 after its insertion into pBluescript II SK^+, and the resulting plasmid was named pLBD-SK. The budC had an open reading frame consisting of 774 bp and encoded 258 amino acids. It was not included in a 2, 3-butanediol operon such as is seen in the case of the meso-BDH gene(budC)of Klebsiella pneumoniae. For the expression of the budC, the deletion plasmid pLBD2-119 was prepared from pLBD-SK. E.coli JM109/pLBD2-119 had higher L-BDH activity than that of Br.saccharolyticum C-1012. The L-BDH appeared as two bands on disc-PAGE. Isopropyl-β-D-thiogalacto-pyranoside(IPTG) influenced the quantity ratio of the electrophoretic isoenzymes of L-BDH from E.coli JM109/pLBD2-119; that is, a higher relative mobility band with weak substrate specificity was abundantly produced by IPTG. The BDH was considered to belong to the short-chain dehydrogenase/reductase GXXXGXG and YXXXK, and it consisted of about 250 amino acids. As a result of a phylogenetic analysis of SDR family enzymes, the BDHs were considered to comprise a clucter independent from the other SDR enzymes.

• 社団法人日本生物工学会の論文
• 1998-09-25

著者

• Ui Sadaharu

  Division of Biotechnology Yamanashi Univ.

• Ohkuma Moriya

  Environmental Molecular Biology Laboratory Riken

• Ohkuma M

  Enviromental Molecular Biology Lab. Riken

• Ohkuma Moriya

  Department Of Agricultural Chemistry The University Of Tokyo

• Ui Sadaharu

  Department Of Applied Chemistry And Biotechnology Faculty Of Engineering Yamanashi University

• Ui Sadaharu

  Division Of Biotechnology Interdisciplinary Graduate School Of Medicine And Engineering University O

• Kudo Toshiaki

  Environmental Molecular Biology Lab. :graduate School Of Integrated Science Yokohama City University

• Kudo Toshiaki

  Laboratory Of Microbiology The Institute Of Physical And Chemical Research(riken)

• Ohkuma Moriya

  Bio-recycle Project Japan Science And Technology Agency:presto Japan Science And Technology Agency:e

• Ui Sadaharu

  Department Of Applied Chemistry & Biotechnology Faculty Of Engineering Yamanashi University

• OTAGIRI Masato

  Department of Applied Chemistry and Biotechnology, Faculty of Engineering, Yamanashi University

• MIMURA Akio

  Department of Applied Chemistry and Biotechnology, Faculty of Engineering, Yamanashi University

• DOHMAE NAOSHI

  Laboratory of Microbiology

• TAKIO KOJI

  Laboratory of Microbiology

• OHKUMA MORIYA

  The Institute of Physical and Chemical Research(RIKEN)

• KUDO TOSHIAKI

  The Institute of Physical and Chemical Research(RIKEN)

• Dohmae N

  Riken Saitama Jpn

• Dohmae Naoshi

  Biomolecular Characterization Team Advanced Technol. Support Div. Riken Advanced Sci. Inst.

• Takio K

  Enviromental Molecular Biology Lab. Riken

• Kudo T

  Inst. Physical And Chemical Res.(riken) Saitama Jpn

• Ohkuma Moriya

  Bio-recycle Project Japan Science And Technology Agency

• Dohmae N

  Riken(inst. Physical And Chemical Res.) Saitama Jpn

• Kudo T

  Riken Saitama Jpn

• Kudo Toshiaki

  International Graduate School Of Arts And Sci. Yokohama City Univ.

• Kudo Toshiaki

  Yokohama City Univ. Kanagawa Jpn

• Mimura A

  Department Of Applied Chemistry And Biotechnology Faculty Of Engineering University Of Yamanashi

• Mimura A

  Hitachi Ltd. Ibaraki Jpn

• Otagiri Masato

  Institute For Protein Research Osaka University

• Mimura A

  Univ. Of Yamanashi Yamanashi Jpn

• Mimura Akiko

  Department Of Applied Chemistry And Biotechnology Faculty Of Engineering University Of Yamanashi

• Kudo Toshiaki

  Environmental Molecular Biology Laboratory Riken

• Mimura Akio

  Department Of Applied Chemistry & Biotechnology Faculty Of Engineering Yamanashi University

• Ui S

  Division Of Biotechnology Interdisciplinary Graduate School Of Medicine And Engineering University O

• KUDO Toshiaki

  The Institute of Physical and Chemical Research

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