文献类型：期刊文献

英文题名：Synthesis of optically pure S-sulfoxide by Escherichia coli transformant cells coexpressing the P450 monooxygenase and glucose dehydrogenase genes

作者：Zhang, Jian-Dong[1];Li, Ai-Tao[1];Yu, Hui-Lei[1];Imanaka, Tadayuki[1,2];Xu, Jian-He[1]

机构：[1]E China Univ Sci & Technol, Lab Biocatalysis & Bioproc, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[2]Ritsumeikan Univ, Coll Life Sci, Dept Biotechnol, Shiga 5258577, Japan

年份：2011

卷号：38

期号：5

起止页码：633

外文期刊名：JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY

收录：;EI(收录号：20242716594017);WOS:【SCI-EXPANDED(收录号:WOS:000289562500006)】；

基金：This work was financially supported by the National Natural Science Foundation of China (grant Nos. 20506037 & 20672037&20902023), Ministry of Science and Technology, P.R. China (grant Nos. 2006AA02Z205 & 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：Coexpression; P450SMO; Glucose dehydrogenase; NADPH regeneration; D-glucose; Sulfoxidation

摘要：A cytochrome P450 monooxygenase (P450SMO) from Rhodococcus sp. can catalyze asymmetric oxygenation of sulfides to S-sulfoxides. However, P450SMO-catalyzed biotransformations require a constant supply of NAD(P)H, the expense of which constitutes a great hindrance for this enzyme application. In this study, we investigated the asymmetric oxygenation of sulfide to S-sulfoxide using E. coli cells, which co-express both the P450SMO gene from Rhodococcus sp. and the glucose dehydrogenase (GDH) gene from Bacillus subtilis, as a catalyst. The results showed that the catalytic performance of co-expression systems was markedly improved compared to the system lacking GDH. When using recombinant E. coli BL21 (pET28a-P450-GDH) whole cell as a biocatalyst, NADPH was efficiently regenerated when glucose was supplemented in the reaction system. A total conversion of 100% was achieved within 12 h with 2 mM p-chlorothioanisole substrate, affording 317.3 mg/L S-sulfoxide obtained. When the initial sulfide concentration was increased to 5 mM, the substrate conversion was also increased nearly fivefold: S-sulfoxide amounted to 2.5 mM (396.6 mg/L) and the ee value of sulfoxide product exceeded 98%. In this system, the effects of glucose concentration and substrate concentration were further investigated for efficient biotransformation. This system is highly advantageous for the synthesis of optically pure S-sulfoxide.