Gene Clonign, Purification, and Characterization of the Highly Thermostable Leucine Dehydrogenase of Bacillus sp.

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概要

• 論文の詳細を見る

• The leucine dehydrogenase (L-leucine : NAD^+ oxidoreductase, deaminating, EC 1.4.1.9) gene from Bacillus sp. DSM730 was cloned into Escherichia coli C600 with a vector plasmid, pBR322. The E.coli cells carrying a recombinant plasmid, pKULD1 (9.5 kb), produced a highly thermostable leucine dehydrogenase. The enzyme from E. coli cells carrying pKULD103,a deletion plasmid (6.5 kb) of pKULD1,was purified to homogeneity from the crude extract of clone cells by only one ion-change chromatography application with a yield of 73%. The leucine dehydrogenase of Bacillus sp. DSM730 is very similar in enzymological properties to those of other bacteria, except for molecular weight and stability. It has a molecular weight of about 280,000 and consists of six subunits identical in molecular weight (47,000). The enzyme is not inactivated by heat treatment at 80℃ for 10 min, and incubation in the pH range of 5.4 to 10.3 at 55℃ for 10 min. The Bacillus sp. DSM730 leucine dehydrogenase is the most thermostable of the leucine dehydrogenases so far purified, and is very useful for structure and stability studies, as well as being applicable to L-leucine production.

• 社団法人日本生物工学会の論文
• 1990-04-25

著者

• MISONO Haruo

  Department of Bioresources Science, Kochi University

• Esaki Nobuyoshi

  Kyoto Univ.

• Esaki Nobuyoshi

  Nstitute For Chemical Research Kyoto University

• Esaki Nobuyoshi

  Laboratory Of Microbial Biochemistry Institute For Chemical Research Kyoto University

• Meguro H

  Tohoku Fukushi University

• Meguro H

  Department Of Applied Biological Chemistry Faculty Of Agriculture Tohoku University

• TANAKA Hidehiko

  Laboratory of Microbiology, Institute for Chemical Research, Kyoto University

• SODA Kenji

  Laboratory of Microbiology, Institute for Chemical Research, Kyoto University

• Soda Kenji

  Laboratory Of Microbial Biochemistry Institute For Chemical Research Kyoto University

• Misono Haruo

  Department Of Bioresources Science Faculty Of Agriculture Kochi University

• Misono Haruo

  Department Of Bioresource Science Kochi University

• NAGATA Shinji

  Department of Internal Medicine, Hiroshima City Asa Hospital

• Tanaka H

  Department Of Bioresources Chemistry Faculty Of Agriculture Okayama University

• Esaki Nobuyoshi

  Institute For Chemical Resarch Kyoto University

• Soda K

  Department Of Biotechnology Kansai University

• Nagata Shinji

  Department Of Applied Biological Chemistry Graduate School Of Agricultural And Life Sciences The Uni

• Tanaka Hidehiko

  Laboratory Of Biochemistry Kyoto College Of Pharmacy

• NAGASAKI Susumu

  Department of Agricultural Chemistry, Kochi University

• Nagasaki S

  Department Of Bioresources Science Kochi University

• Moriyama Hideaki

  Japan Synchrotron Radiation Research Institute (JASRI)

• Nagasaki Susumu

  Department Of Agricultural Chemistry Faculty Of Agriculture Kochi University

• NAGATA Shinji

  Department of Agricultural Chemistry, Kochi University

• MISONO Haruo

  Department of Agricultural and Biological Chemistry, Kochi University

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