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

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Journal of Industrial Microbiology & Biotechnology

Abstract

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.

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Acknowledgments

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).

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Authors and Affiliations

  1. Laboratory of Biocatalysis and Bioprocessing, State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Road, 200237, Shanghai, People’s Republic of China

    Jian-Dong Zhang, Ai-Tao Li, Hui-Lei Yu, Tadayuki Imanaka & Jian-He Xu

  2. Department of Biotechnology, College of Life Sciences, Ritsumeikan University, Shiga, 525-8577, Japan

    Tadayuki Imanaka

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  1. Jian-Dong Zhang
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  2. Ai-Tao Li
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  3. Hui-Lei Yu
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  4. Tadayuki Imanaka
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Correspondence to Jian-He Xu.

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Zhang, JD., Li, AT., Yu, HL. et al. Synthesis of optically pure S-sulfoxide by Escherichia coli transformant cells coexpressing the P450 monooxygenase and glucose dehydrogenase genes. J Ind Microbiol Biotechnol 38, 633–641 (2011). https://doi.org/10.1007/s10295-010-0809-3

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  • DOI: https://doi.org/10.1007/s10295-010-0809-3

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