Identification and characterization of another 4-nitrophenol degradation gene cluster, nps, in Rhodococcus sp. strain PN1(ENVIRONMENTAL BIOTECHNOLOGY)
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概要
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4-Nitrophenol (4-NP) is a toxic compound formed in soil by the hydrolysis of organophosphorous pesticides, such as parathion. We previously reported the presence of the 4-NP degradation gene cluster (nphRA1A2) in Rhodococcus sp. strain PN1, which encodes a two-component 4-NP hydroxylase system that oxidizes 4-NP into 4-nitrocatechol. In the current study, another gene cluster (npsC and npsRA2A1B) encoding a similar 4-NP hydroxylase system was cloned from strain PN1. The enzymes from this 4-NP hydroxylase system (NpsA1 and NpsA2) were purified as histidine-tagged (His-) proteins and then characterized. His-NpsA2 showed NADH/FAD oxidoreductase activity, and His-NpsA1 showed 4-NP oxidizing activity in the presence of His-NpsA2. In the 4-NP oxidation using the reconstituted enzyme system (His-NpsA1 and His-NpsA2), hydroquinone (35% of 4-NP disappeared) and hydroxyquinol (59% of 4-NP disappeared) were detected in the presence of ascorbic acid as a reducing reagent, suggesting that, without the reducing reagent, 4-NP was converted into their oxidized forms, 1,4-benzoquinone and 2-hydroxy-1,4-benzoquinone. In addition, in the cell extract of recombinant Escherichia coil expressing npsB, a typical spectral change showing conversion of hydroxyquinol into maleylacetate was observed. These results indicate that this nps gene cluster, in addition to the nph gene cluster, is also involved in 4-NP degradation in strain PN1.
著者
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Takeo Masahiro
Department of Materials Science and Chemistry, Graduate School of Engineering, University of Hyogo
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Kato Dai-ichiro
Department of Materials Science and Chemistry, Graduate School of Engineering, University of Hyogo
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Negoro Seiji
Department of Materials Science and Chemistry, Graduate School of Engineering, University of Hyogo
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YAMAMOTO Kenta
Department of Cardiovascular Dynamics, Advanced Medical Engineering Center, National Cardiovascular
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Negoro Seiji
Department Of Materials Science And Chemistry Graduate School Of Engineering University Of Hyogo
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Negoro Seiji
Department Of Biotechnology Faculty Of Engineering Osaka University
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Takeo Masahiro
Department Of Materials Science And Chemistry Graduate School Of Engineering University Of Hyogo
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Takeo Masahiro
Department Of Applied Chemistry Himeiji Institute Of Technology
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Negoro Seiji
Graduate School Of Engineering Univ. Of Hyogo
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Yasuhira Kengo
Department of Materials Science and Chemistry, Graduate School of Engineering, University of Hyogo
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Takeo Masahiro
兵庫県立大学 工学研究科物質系工学専攻
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Negoro Seiji
兵庫県立大学 工学研究科物質系工学専攻
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Yasuhira Kengo
Department Of Materials Science And Chemistry Graduate School Of Engineering University Of Hyogo
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Kato Dai-ichiro
Graduate School Of Engineering University Of Hyogo
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Kato Dai-ichiro
Department Of Materials Science And Chemistry Graduate School Of Engineering University Of Hyogo
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Yamamoto Kenta
Department Of Materials Science And Chemistry Graduate School Of Engineering University Of Hyogo
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Nishimura Munehiro
Department Of Materials Science And Chemistry Graduate School Of Engineering University Of Hyogo
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