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Enhanced selenocysteine biosynthesis for seleno-methylselenocysteine production in Bacillus subtilis

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Abstract

Seleno-methylselenocysteine (SeMCys) is an effective component for selenium supplementation with anti-carcinogenic potential and can ameliorate neuropathology and cognitive deficits. In this study, we aimed to engineer Bacillus subtilis 168 for the microbial production of SeMCys. First, the accumulation of intracellular selenocysteine (SeCys) as the precursor of SeMCys was enhanced through overexpression of serine O-acetyltransferase, which was desensitized against feedback inhibition by cysteine. Next, the S-adenosylmethionine (SAM) synthetic pathway was optimized to improve methyl donor availability through expression of S-adenosylmethionine synthetase. Further, SeMCys was successfully produced through expression of the selenocysteine methyltransferase in SeCys and SAM-producing strain. The increased expression level of selenocysteine methyltransferase benefited the SeMCys production. Finally, all the heterologous genes were integrated into the genome of B. subtilis, and the strain produced SeMCys at a titer of 18.4 μg/L in fed-batch culture. This is the first report on the metabolic engineering of B. subtilis for microbial production of SeMCys and provides a good starting point for future pathway engineering to achieve the industrial-grade production of SeMCys.

Key points

• Expression of the feedback-insensitive serine O-acetyltransferase provided B. subtilis the ability of accumulating SeCys.

• SAM production was enhanced through expressing S-adenosylmethionine synthetase in B. subtilis.

• Expression of selenocysteine methyltransferase in SeCys and SAM-accumulating strain facilitated SeMCys production.

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Data availability

The source data supporting the findings of this study are available within the paper.

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Acknowledgements

We are grateful to Prof. Long Liu (Jiangnan University, Wuxi, China) for generously providing the plasmids.

Funding

This study was financially supported by the National Natural Science Foundation of China (nos. 21808005 and PXM2019_014213_000007), Beijing Municipal Education Commission (no. KM201910011005), and the Open Project Program from Key Laboratory of Cleaner Production and Integrated Resource Utilization of China, Beijing Technology and Business University (BTBU) (no. CP-2020-YB12).

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Authors and Affiliations

  1. Beijing Advanced Innovation Center for Food Nutrition and Human Health, Beijing Technology and Business University, Fucheng RD 11, Beijing, 100048, China

    Xian Yin, Yu Zhou, Hulin Yang, Yonghong Liao & Fenghuan Wang

  2. School of Light Industry, Beijing Technology and Business University, Fucheng RD 11, Beijing, 100048, China

    Xian Yin, Yu Zhou, Hulin Yang, Yonghong Liao & Fenghuan Wang

  3. Biological Defense Department, Institute of Chemical Defence, Zhongxin RD 1, Beijing, 102205, China

    Tengbo Ma

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  1. Xian Yin
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  2. Yu Zhou
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  3. Hulin Yang
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Contributions

X.Y. and F.W. designed the experiments. Y.Z. and H.Y. constructed the plasmids. X.Y., Y.Z., and T.M. performed the other experiments. X.Y. and Y.L. analyzed the data. All authors discussed the results. X.Y. prepared the manuscript. All authors contributed to the revision of the manuscript.

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Correspondence to Fenghuan Wang.

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Yin, X., Zhou, Y., Yang, H. et al. Enhanced selenocysteine biosynthesis for seleno-methylselenocysteine production in Bacillus subtilis. Appl Microbiol Biotechnol 107, 2843–2854 (2023). https://doi.org/10.1007/s00253-023-12482-8

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