Purification and Some Properties of Glutamine Synthetases from Bifidobacteria(Microbiology & Fermentation Industry)

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概要

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• Glutamine synthetases [L-gultamate; ammonia ligase (ADP forming), EC 6.3.1.2] were purified, respectively, 570-, 220- and 80-fold with 12, 5 and 0.5% yields from cell-free extracts of Bifidobacterium bifidum a, B. breve 203 and B. pseudolongum a. The purification procedure consisted of cell disruption, ammonium sulfate fractionation and column chromatographies on DEAE-cellulose, Sepharose 6B and hydroxylapatite. The purified enzymes were judged to be homogeneous on polyacrylamide disc gel electrophoresis. they had similar molecular weights of about 500,000. The optimum pH was 6.0〜6.5 for their biosynthetic reactions with 30 mM Mg^<2+>, and 6.5 for their γ-gultamylhydroxamate forming (transferring) reactions. the Km values for L-glutamate, ammonia and ATP in their biosynthetic reactions were, respectively, 18〜25 mM, 0.56〜0.69 mM and 1.5〜2.0 mM with 30 mM Mg^<2+> as a cofactor. When 7.5 mM Mn^<2+> was substituted for 30 mM Mg^<2+>, the Km for L-glutamate decreased by 8.6〜23-fold. The optimum pH for the biosynthetic reactions and the susceptibility of the enzymes to some metabolites also varied with the kind of divalent cation added.

• 社団法人日本農芸化学会の論文
• 1987-02-23

著者

• TACHIKI TAKASHI

  Department of Food Science and Technology, Kyoto University

• TOCHIKURA TATSUROKURO

  Department of Food Science and Technology, Kyoto University

• Tochikura Tatsurokuro

  Department Of Agricultural Chemistry Kyoto University

• Tachiki Takashi

  Department Of Chemistry Ritsumeikan University

• HATANAKA Masayuki

  Department of Food Science and Technology, Kyoto University

• Tachiki Takashi

  Faculty Of Home Economics Kobe Women's University

• Hatanaka Masayuki

  Department Of Food Science And Technology Kyoto University

• FURUKAWA Ryoko

  Department of Food Science and Technology, Faculty of Agriculture, Kyoto University

• Tachiki Takashi

  Department Of Bioscience & Technology Faculty Of Science & Engineering Ritsumeikan Universit

• Furukawa Ryoko

  Department Of Food Science And Technology Faculty Of Agriculture Kyoto University

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