NADP-Specific Glutamate Dehydrogenase from Alkaliphilic Bacillus sp. KSM-635: Purification and Enzymatic Properties
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概要
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An NADP-specific glutamate dehydrogenase [L-glutamate: NADP^+ oxidoreductase (deaminating), EC 1.4.1.4] from alkaliphilic Bacillus sp. KSM-635 was purified 5840-fold to homogeneity by a several-step procedure involving Red-Toyopearl affinity chromatography. The native protein, with an isoelectric point of pH 4.87, had a molecular mass of approximately 315 kDa consisting of six identical subunits each with a molecular mass of 52 kDa. The pH optima for the aminating and deaminating reactions were 7.5 and 8.5, respectively. The optimum temperature was around 60℃ for both. The purified enzyme had a specific activity of 416 units/mg protein for the aminating reaction, being over 20-fold greater than that for deaminating reaction, at the respective pH optima and at 30℃. The enzyme was specific for NADPH (K_m 44 μM), 2-oxoglutarate (K_m 3.13 mM), NADP^+ (K_m 29 μM), and L-glutamate (K_m 6.06 mM). The K_m for NH_4Cl was 5.96 mM. The enzyme could be stored without appreciable loss of enzyme activity at 5℃ for half a year in phosphate buffer (pH 7.0) containing 2 mM 2-mercaptoethanol, although the enzyme activity was abolished within 20 h by freezing at -20℃.
- 社団法人日本農芸化学会の論文
- 1996-11-23
著者
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Hatada Y
Japan Agency For Marine‐earth Sci. And Technol. (jamstec) Yokosuka Jpn
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Hatada Yuji
Tochigi Research Laboratories Of Kao Corporation
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Hayashi Y
Research Institute For Food Science Kyoto University
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Koike K
Faculty Of Pharmaceutical Sciences Toho University
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Ito S
Extremobiosphere Research Center Of Japan Agency For Marine-earth Science And Technology (jamstec):c
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ITO Susumu
Tochigi Research laboratories, Kao Corporation
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Koike Kenzo
Tochigi Research Laboratories, Kao Corporation
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Kobayashi Tohru
Extremobiosphere Research Center of Japan Agency for Marine-Earth Science and Technology (JAMSTEC)
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KOBAYASHI Tohru
Tochigi Research Laboratories of Kao Corporation
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HAKAMADA Yoshihiro
Tochigi Research Laboratories of Kao Corporation
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YOSHIMATSU Tadashi
Tochigi Research Laboratories of Kao Corporation
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Kobayashi T
Extremobiosphere Research Center Of Japan Agency For Marine-earth Science And Technology (jamstec)
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Koike K
Tochigi Research Laboratories Kao Corporation
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Ito S
National Industrial Research Institute
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Hayashi Y
Department Of Medicinal Chemistry Center For Frontier Research In Medicinal Science Kyoto Pharmaceut
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Sugimoto Isamu
Division Of Pathological Biochemistry Department Of Biomedical Sciences School Of Life Sciences Facu
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Kobayashi Tohru
Tochigi Research Laboratories Kao Corporation
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Ito Susumu
Tochigi Research Laboratories Kao Corporation
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