Purification and Properties of Malate Dehydrogenase from Ammonia-Oxidizing Chemoautotrophic Nitrosomonas europaea ATCC 25978
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概要
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Ammonia-oxidizing Chemoautotrophic Nitrosomonas europaea exhibited remarkable oxaloacetate-reducing activity with cosubstrates. During purification on DEAE-Sepharose CL-6B, a fraction containing malate dehydrogenase [EC 1.1.1.37], i,e, MDH, was found. The enzyme protein was further purified to electrophoretic homogeneity by ion exchange chromatography and gel filtration. The molecular weight of the enzyme was estimated to be about 110,000 by gel filtration, suggesting that the enzyme consisted of two different subunits (α: 25,000, β: 36,000), as demonstrated by SDS-PAGE. The purified enzyme required eiter NADH or NADPH as a coenzyme. The isoelectric points on NADH or NADPH-linked MDH was pH 6.75. The pH and temperature optima on the NADH-linked enzyme activity (NADPH-linked enzyme activity) were about 7.0 (4.5) and 50℃ (45-50℃). The enzyme was stable up to 50℃ at both enzyme activities. It was active specifically for oxaloacetate but not for any other organic acids. It was activated by CuSO_4 and CoCl_2 at both enzyme activities. The activity of NADPH-linked MDH was strongly inhibited by PbCl_2, while that of NADH-linked MDH was not.
- 1992-12-01
著者
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Takahashi Reiji
Department Of Agricultural And Biological Chemistry College Of Bioresource Sciences Nihon University
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Tokuyama Tatsuaki
Department Of Agricultural And Biological Chemistry College Of Bioresource Sciences Nihon University
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Kobayashi Makoto
Department Of Gastroenterology And Hepatology Division Of Clinical Medicine And Biomedical Science I
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Kodama Masaaki
Department Of Agricultural Chemistry College Of Agriculture And Veterinary Medicine Nihon University
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IGAI Kazuo
Department of Agricultural Chemistry, College of Agriculture and Veterinary Medicine, Nihon University
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Igai Kazuo
Department Of Agricultural Chemistry College Of Agriculture And Veterinary Medicine Nihon University
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Kobayashi Makoto
Department Of Agricultural Chemistry College Of Agriculture And Veterinary Medicine Nihon University
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