Expression and characterization of styrene monooxygenases of Rhodococcus sp. ST-5 and ST-10 for synthesizing enantiopure (S)-epoxides

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• 作者：Hiroshi Toda (1)
  Ryouta Imae (1)
  Tomoko Komio (2)
  Nobuya Itoh (1) nbito@pu-toyama.ac.jp
• 关键词：Rhodococcus sp. – ; Styrene monooxygenase – ; Flavin oxidoreductase – ; Epoxidation – ; (S) ; epoxides – ; Biocatalysis
• 刊名：Applied Microbiology and Biotechnology
• 出版年：2012
• 出版时间：October 2012
• 年：2012
• 卷：96
• 期：2
• 页码：407-418
• 全文大小：2.5 MB
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• 作者单位：1. Department of Biotechnology, Biotechnology Research Center, Faculty of Engineering, Toyama Prefectural University, 5180 Kurokawa, Imizu, Toyama, 939-0398 Japan2. Namerikawa High School, 45 Kashimacho, Namerikawa, Toyama, 936-8507 Japan
• ISSN：1432-0614

文摘

Styrene monooxygenase (StyA, SMOA)- and flavin oxidoreductase (StyB, SMOB)-coding genes of styrene-assimilating bacteria Rhodococcus sp. ST-5 and ST-10 were successfully expressed in Escherichia coli. Determined amino acid sequences of StyAs and StyBs of ST-5 and ST-10 showed more similarity with those of Pseudomonas than with self-sufficient styrene monooxygenase (StyA2B) of Rhodococcus. Recombinant enzymes were purified from E. coli cells as functional proteins, and their properties were characterized in detail. StyBs (flavin oxidoreductase) of strains ST-5 and ST-10 have similar enzymatic properties to those of Pseudomonas, but StyB of strain ST-10 exhibited higher temperature stability than that of strain ST-5. StyAs of strains ST-5 and ST-10 catalyzed the epoxidation of vinyl side-chain of styrene and its derivatives and produced (S)-epoxides from styrene derivatives and showed high stereoselectivity. Both StyAs showed higher specific activity on halogenated styrene derivatives than on styrene itself. Additionally, the enzymes could catalyze the epoxidation of short-chain 1-alkenes to the corresponding (S)-epoxides. Aromatic compounds including styrene, 3-chlorostyrene, styrene oxide, and benzene exhibited marked inhibition of SMO reaction, although linear 1-alkene showed no inhibition of SMO activity at any concentration.