文献类型：期刊文献

英文题名：Sequence analysis and heterologous expression of a new cytochrome P450 monooxygenase from Rhodococcus sp for asymmetric sulfoxidation

作者：Zhang, Jian-Dong;Li, Ai-Tao;Yang, Yi;Xu, Jian-He[1]

机构：[1]E China Univ Sci & Technol, State Key Lab Bioreactor Engn, Lab Biocatalysis & Bioproc, Shanghai 200237, Peoples R China

年份：2010

卷号：85

期号：3

起止页码：615

外文期刊名：APPLIED MICROBIOLOGY AND BIOTECHNOLOGY

收录：;EI(收录号：20100312639418);WOS:【SCI-EXPANDED(收录号:WOS:000273351300020)】；

基金：This work was financially supported by the National Natural Science Foundation of China (grant nos. 20506037 and 20672037), Ministry of Science and Technology, P. R. China (grant nos. 2006AA02Z205 and 2007AA02Z225) and China National Special Fund for State Key Laboratory of Bioreactor Engineering (grant no. 2060204).

语种：英文

外文关键词：Cytochrome P450 monooxygenase; Rhodococcus sp; Cloning; Expression; Asymmetric sulfoxidation

摘要：In this study, a 3.7-kb DNA fragment was cloned from Rhodococcus sp. ECU0066, and the sequence was analyzed. It was revealed that the largest one (2,361 bp) of this gene fragment encodes a protein consisting of 787 amino acids, with 73% identity to P450RhF (accession number AF45924) from Rhodococcus sp. NCIMB 9784. The gene of this new P450 monooxygenase (named as P450SMO) was successfully expressed in Escherichia coli BL21 (DE3), and the enzyme was also purified and characterized. In the presence of reduced nicotinamide adenine dinucleotide phosphate, the enzyme showed significant sulfoxidation activity towards several sulfides, with (S)-sulfoxides as the predominant product. The p-chlorothioanisole, p-fluorothioanisole, p-tolyl methyl sulfide, and p-methoxythioanisole showed relatively higher activities than the other sulfides, but the stereoselectivity for p-methoxythioanisole was much lower. The optimal activity of the purified enzyme toward p-chlorothioanisole occurred at pH 7.0 and 30A degrees C. The current study is the first to report a recombinant cytochrome P450 enzyme of Rhodococcus sp. which is responsible for the asymmetric oxidation of sulfides. The new enzymatic activity of P450SMO on the above compounds makes it an attractive biocatalyst for asymmetric synthesis of enantiopure sulfoxides.