A Non-NadB Type L-Aspartate Dehydrogenase from Ralstonia eutropha Strain JMP134 : Molecular Characterization and Physiological Functions

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概要

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• We report the molecular characterization and physiological function of a novel L-aspartate dehydrogenase (AspDH). The purified enzyme was a 28-kDa dimeric protein, exhibiting high catalytic activity for L-aspartate (L-Asp) oxidation using NAD and/or NADP as cofactors. Quantitative real-time PCR analysis indicated that the genes involved in the AspDH gene cluster, poly-3-hydroxyalkanoate (PHA) biosynthesis, and the TCA cycle were substantially induced by L-Asp in wild-type cells. In contrast, expression of the aspartase and aspartate aminotransferase genes was substantially induced in the AspDH gene knockout mutant (ΔB3576) but not in the wild type. GC-MS analyses revealed that the wild-type strain synthesized poly-3-hydroxybutyrate from fructose or L-Asp, whereas the ΔB3576 mutant did not synthesize PHA from L-Asp. AspDH gene cluster products might be involved in the biosynthesis of the PHA precursor, revealing that AspDH was a non-NadB type enzyme, and thus entirely different from the previously reported NadB type enzymes working in NAD biosynthesis.

• 社団法人 日本農芸化学会の論文
• 2011-08-23

著者

• ISHIDA Manabu

  Department of High Energy Astrophysics, Institute of Space and Astronautical Science (ISAS), Japan A

• Ishikawa Tomohiro

  Department Of Agricultural Chemistry School Of Agriculture Nagoya University

• Shibata Hiroyuki

  Institute For Chemical Research Kyoto University

• Ashida Hiroyuki

  Mol. Gen. Res. Inst. Shimane Univ.

• Ashida Hiroyuki

  Institute For Molecular Genetics Shimane University

• Ishikawa T

  Fac. Of Life And Environmental Sci. Shimane Univ.

• Shibata H

  Dept. Life Environ. Sci. Shimane Univ.

• Sawa Yoshihiro

  Dept. Life Environ. Sci. Shimane Univ.

• LI Yinxia

  Department of Life Science and Biotechnology, Faculty of Life and Environmental Science, Shimane Uni

• Sakamoto Yoko

  Department Of Food Science And Technology College Of Bioresouce Sciences Nihon University

• Ishikawa Takahiro

  Department Of Clinical Cell Biology And Medicine Chiba University Graduate School Of Medicine

• Shibata Hitoshi

  Department Of Applied Biological Science Faculty Of Agriculture Shimane University

• Sawa Yoshihiro

  Department Of Life Science And Biotechnology Faculty Of Life And Environmental Science Shimane Unive

• Ashida Hiroyuki

  Department Of Molecular And Functional Genomics Center For Integrated Research In Science Shimane Un

• Li Yinxia

  Department Of Life Science And Biotechnology Faculty Of Life And Environmental Science Shimane Unive

• Ishida Manabu

  Department Of Life Science And Biotechnology Faculty Of Life And Environmental Science Shimane University

• Sawa Yoshihiro

  Department Of Applied Bioscience And Biotechnology Faculty Of Life And Environmental Science Shimane University

• Ishikawa Takahiro

  Department of Applied Bioscience and Biotechnology, Faculty of Life and Environmental Science, Shimane University

• SHIBATA Hitoshi

  Department of Agricultural Chemistry, Faculty of Agriculture, Shinshu University

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