Purification and Characterization of Nicotinic Acid Dehydrogenase from Pseudomonas fluorescens TN5
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概要
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Membrane-bound nicotinic acid dehydrogenase, an enzyme that catalyzes the formation of 6-hydroxynicotinic acid from nicotinic acid, was solubilized with Triton X-100,and then purified 126-fold with an 11.1% overall recovery from nicotinic acid-induced cells of Pseudomonas fluorescens TN5. The purified enzyme appeared to be homogeneous from analysis by polyacrylamide gel electrophoresis. The enzyme had a molecular mass of approximately 80 kDal and consisted of one subunit. Some electron acceptors, such as phenazine methosulfate, K_3Fe(CN)_6 and nitro blue tetrazolium, acted as electron acceptors. The purified enzyme catalyzed the hydroxylation of nicotinic acid to 6-hydroxynicotinic acid at a rate of 672 μmol min^<-1> mg^<-1> of protein at 35℃. It also catalyzed the hydroxylation of pyrazinecarboxylic acid, 3-pyridinesulfonic acid, and 3-cyanopyridine. The purified enzyme exhibited an optimum pH of 8.3,and was sensitive to thiol regents such as HgCl_2 and p-chloromercuribenzoate. A reduction in the amount of cytochrome c-like component in the respiratory particles was observed during the hydroxylation reaction of nicotinic acid. Thus, nicotinic acid dehydrogenase appeared to be linked to the cytochrome respiratory chain in the cells of P. fluorescens TN5.
- 公益社団法人日本生物工学会の論文
- 1994-07-25
著者
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Yamane T
Laboratory Of Molecular Biotechnology Graduate School Of Bioagricultural Sciences Nagoya University:
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Yamane Tsuneo
Laboraotory Of Bioreaction Engineering Department Of Food Science And Technology Faculty Of Agricult
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Yamane Tsuneo
Laboratory Of Bioreaction Engineering Department Of Food Science And Technology School Of Agricultur
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Yamane T
Laboratory Of Molecular Biotechnology Graduate School Of Biological & Agricultural Sciences Nago
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Yamane T
Laboratory Of Bioreaction Engineering Department Of Food Science And Technology School Of Agricultur
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Nagasawa Toru
Department Of Agricultural Chemistry Kyoto University
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Yamane Tsuneo
Department Of Applied Biological Sciences Faculty Of Agriculture Nagoya University
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Yamane Tsuneo
Laboratory Of Molecular Biotechnology Graduate School Of Bio- And Agro-sciences Nagoya University
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HURH BYUNGSERK
Department of Food Science and Technology, Faculty of Agriculture, Nagoya University
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Hurh Byungserk
Department Of Food Science And Technology Nagoya University
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