An NADP-Glutamate Dehydrogenase from the Green Alga Bryopsis maxima. Purification and Properties
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概要
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NADP-glutamate dehydrogenase (EC 1.4.1.4; NADP-GDH) was purified to electrophoretic homogeneity from the multinuclear-unicellular green marine alga in Siphonales, Bryopsis maxima, and its properties were examined. M_r of the undenatured enzyme was 280 kDa, and the enzyme is thought to be a hexamer of 46 kDa subunit protein. Optimum pHs for the reductive amination and oxidative deamination were 7.5 and 8.2-9.0 respectively. The enzyme displayed NADPH/NADH-specific activities with a ratio of 18 : 1. Apparent K_m values for 2-oxoglutarate, ammonia, NADPH, glutamate and NADP^+ were 3.0, 2.2, 0.03, 3.2 and 0.01 mM respectively. The enzymochemical characteristics of the GDH were studied and compared to those of other species. The B. maxima GDH was insensitive to 5 mM Ca^<2+> and to 1 mM EDTA in contrast to higher plant NAD-GDHs. Chemical modifications with DTNB and pCMBS suggested that cysteine residues are essential for the enzymatic activity as in other species GDHs. The GDH was not affected by 1 mM purine nucleotides, suggesting that the enzyme is not allosteric, in contrast to animal NAD(P)-GDHs and fungal NAD-GDHs.
- 日本植物生理学会の論文
著者
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OKADA Mitsumasa
Department of Chemical Engineering, Graduate School of Engineering, Hiroshima University, Japan
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ITAGAKI Tadashi
Department of Microbiology, College of Pharmacy, Nihon University
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Okada Mitsumasa
Department Of Biomolecular Science Toho University
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Okada Mitsumasa
Department Of Biomolecular Science Faculty Of Science Toho University
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Itagaki T
Nippon Bio-rad Laboratories Diagnostics Group
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Wiskich Joseph
Department Of Botany University Of Adelaide
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Inokuchi Ritsuko
Departments of Microbiology, Nihon University School of Dentistry at Matsudo
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Okada M
Department Of Biomolecular Science Faculty Of Science Toho University
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Itagaki Tadashi
Nippon Bio-Rad Laboratories, Diagnostics Group
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Nakayama Katsumi
Department of Biomolecular science, Faculty of science, Toho University
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Nakayama K
Toho Univ. Chiba Jpn
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Nakayama Katsumi
Department Of Biomolecular Science Toho University
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Nakayama Katsumi
Department Of Biology Faculty Of Science Toho University
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Inokuchi R
Toho Univ. Chiba Jpn
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Inokuchi Ritsuko
Department Of Biomolecular Science Faculty Of Science Toho University
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Okada Mitsumasa
Department Of Biochemistry Faculty Of Science University Of Tokyo
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