Purification and Characterization of Dibenzothiophene (DBT) Sulfone Monooxygenase, an Enzyme Involved in DBT Desulfurization, from Rhodococcus erythropolis D-1
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概要
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Dibenzothiophene (DBT), a model of organic sulfur compound in petroleum, is microbially desulfurized to 2-hydroxybiphenyl by Rhodococcus erythropolis D-1. Three desulfurization (Dsz) enzymes-DszC, A, and B-and flavin reductase are involved in sulfur-specific DBT desulfurization. In this study, DszA was purified, characterized, and crystallized from R.erythropolis D-1. DszA, DBT sulfone monooxygenase, is the second enzyme in microbial DBT desulfurization metabolism and catalyzes the conversion of DBT sulfone to 2'-hydroxybiphenyl 2-sulfinic acid in the presence of flavin reductase with cleavage of the carbon-sulfur bond in the DBT skeleton. Using anion-exchange column chromatography, the four enzyme fractions responsible for DBT desulfurization were separated, and DszA was then purified to homogeneity. Polygonal crystals of DszA were observed within a week. DszA was found to have a molecular mass of 97 kDa and to consist of two subunits with identical masses of 50 kDa. The N-terminal amino acid sequence of the purified DszA completely coincided with the deduced amino acid sequence for dszA of R.erythropolis IGTS8 except for a methionine residue at the latter N-terminal. The optimal temperature and pH for DszA activity were 35℃ and about 7.5. The activity of the enzyme was inhibited by Mn^<2+> Ni^<2+> 2, 2'-bipyridine, and 8-quinolinol, suggesting that a metal might be involved in its activity. DszA acted on not only DBT sulfone but also on dibenz[c, e][1, 2]oxathiin 6-oxide and dibenz[c, e][1, 2]oxathiin 6, 6-dioxide. Dihydroxybiphenyl was formed from the latter two substrates.
- 公益社団法人日本生物工学会の論文
- 1999-12-25
著者
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Oshiro Takashi
Department Of Aquatic Biosciences Tokyo University Of Fisheries
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Kojima Tatsuya
Department Of Biotechnology Tottori University
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Ohshiro Takashi
Department Of Biotechnology Faculty Of Engineering Tottori University
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Izumi Yoshikazu
Department Of Agricultural Chemistry Kyoto University
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TORII KUNIAKI
Department of Biotechnology, Tottori University
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KAWASOE HIROSHI
Department of Biotechnology, Tottori University
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Torii Kuniaki
Department Of Biotechnology Tottori University
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Kawasoe Hiroshi
Department Of Biotechnology Tottori University
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Oshiro Takashi
Department of Biotechnology, Tottori University
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OSHIRO Takashi
Department of Agricultural Chemistry, Kyoto University
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Kojima Tatsuya
Department of Applied Chemistry, Nagoya Institute of Technology, Gokiso, Showa, Nagoya 466-8555, Japan
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