Purification and Some Properties of Glutaminase from Pseudomonas nitroreducens IFO 12694

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概要

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• Glutaminase (EC 3.5.1.2) was isolated from Pseudomonas nitroreducens IFO 12694 grown on 0.6% sodium glutamate as a nitrogen source (325-fold purification, 13% yield). The molecular weight of the enzyme was estimated to be 40,000 by gel filtration and SDS-gel electrophoresis. The enzyme hydrolyzed glutamine optimally at pH 9, and its K_m was 6.5mM. D-Glutamine, γ-glutamyl p-nitroanilide, γ-glutamylmethylamide, γ-glutamylethylamide (theanine), and glutathione showed respectively 107, 85, 78, 74, and 82% reactivity of glutamine. Zn^<2+>, Ni^<2+>, Cd^<2+>, Co^<2+>, Fe^<2+>, and Cu^<2+> repressed the enzyme activity strongly. Glutaminase formed γ-glutamylhydroxamate in the reaction mixture containing glutamine and hydroxylamine (transferring reaction). The optimum pH of the transferring reaction was 7-8, and the K_m for glutamine and hydroxylamine were 4mM and 120 mM, respectively. γ-Glutamyl derivatives hydrolyzable by glutaminase showed reactivity for the transferring reaction. Methylamine or ethylamine was replaceable for hydroxylamine with 3 or 8% reactivity. The effect of divalent cations was not so striking as in the hydrolyzing reaction.

• 1996-07-23

著者

• YAMADA Takeshi

  Department of Cardiology, Kouseikai Takeda Hospital

• Hamada Yuki

  Department Of Food Science And Technology Tokyo University Of Fisheries

• UEDA Masashi

  Department of Patho-Functional Bioanalysis, Graduate School of Pharmaceutical Sciences, Kyoto Univer

• TACHIKI TAKASHI

  Department of Food Science and Technology, Kyoto University

• NAEMURA Yasuki

  Department of Bioscience and Biotechnology, Faculty of Science and Engineering, Ritsumeikan Universi

• IMAMURA Nobuo

  Department of Bioscience and Biotechnology, Faculty of Science and Engineering, Ritsumeikan Universi

• HAMADA Yu-ichi

  Department of Bioscience and Biotechnology, Faculty of Science and Engineering, Ritsumeikan Universi

• SHIODE Ju-ichi

  Department of Bioscience and Biotechnology, Faculty of Science and Engineering, Ritsumeikan Universi

• Ueda Masashi

  Department Of Anesthesiology Kyoto Prefectural University Of Medicine

• Hamada Yu-ichi

  Department Of Food Science And Technology Tokyo University Of Fisheries

• Shiode Ju-ichi

  Department Of Bioscience And Biotechnology Faculty Of Science And Engineering Ritsumeikan University

• Naemura Yasuki

  Department Of Bioscience And Biotechnology Faculty Of Science And Engineering Ritsumeikan University

• Yamada Takao

  Department Of Agricultural Chemistry Faculty Of Agriculture Niigata University

• Tachiki T

  Department Of Food Science And Technology Faculty Of Agriculture Kyoto University

• Imamura Nobuo

  Department Of Bioscience And Biotechnology Faculty Of Science And Engineering Ritsumeikan University

• Yamada T

  Laboratory Of Applied Bioorganic Chemistry Division Of Life Science Graduate School Of Agricultural

• Tachiki Takashi

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

• Ueda Masashi

  Department Of Anesthesiology And Reanimatology Faculty Of Medical Sciences University Of Fukui

• Yamada Takeshi

  Department Of Anesthesiology Iida Municipal Hospital

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