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Enantioselectivity and key residue of Herbaspirillum huttiense monooxygenase in asymmetric epoxidation of styrenes

  • Biotechnologically Relevant Enzymes and Proteins
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Abstract

Styrene monooxygenases (SMOs) are powerful enzymes for the synthesis of enantiopure epoxides, but these SMOs have narrow substrate spectra, and the residues in controlling enantioselectivity of SMOs remains unclear. A monooxygenase from Herbaspirillum huttiense (HhMO) was found to have excellent enantioselectivities and diastereoselectivities in the epoxidation of unconjugated terminal alkenes. Here we found that HhMO could also transfer styrene into styrene epoxide with 75% ee, and it could also catalyze the epoxidation of styrene derivatives into the corresponding epoxides with enantioselectivities up to 99% ee. Meanwhile, site 199 in the substrate access channel of HhMO was found to play an important role in the controlling enantioselectivity of the epoxidation. The E199L variant catalyzed the epoxidation of styrene with > 99% ee. The identification of critical residue that affects the enantioselectivity of SMOs would thus be valuable for creating efficient monooxygenases for the preparation of essential enantiopure epoxides.

Key points

• Bioexpoxidation of both conjugated and unconjugated alkenes by HhMO with excellent enantioselectivities.

• Gating residue 199 played an essential role in controlling the enantioselectivity of SMO.

• HhMO E199L catalyzed the epoxidation of styrenes with up to > 99% ee.

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Data availability

All data generated or analyzed during this study are included in this published article (and its supplementary information files).

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Acknowledgements

We thank the NMR center in the College of Science at Henan Agricultural University for taking NMR spectra.

Funding

This study was funded by the National Natural Science Foundation of China (grant nos. 32171472, 31900876), the Natural Science Foundation of Henan Province (grant no. 202300410218), the Key Scientific Research Projects for Higher Education of Henan Province (grant no. 192102110163), and the High-level Talent Scientific Research Startup Fund Program of Henan University of Technology (2019BS020).

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Author notes

  1. Shuang Dong and Donglin Fan These authors contributed equally to the work.

Authors and Affiliations

  1. College of Life Sciences, Henan Agricultural University, 95 Wenhua Road, Zhengzhou, 450002, China

    Shuang Dong, Donglin Fan, Qian Liu, Yinyin Meng, Xinyu Liu, Sen Yang, Hui Lin & Hongge Chen

  2. College of Biological Engineering, Henan University of Technology, 100 Lianhua Street, Zhengzhou, 450001, Henan, China

    Na Li

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  1. Shuang Dong
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  2. Donglin Fan
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Contributions

SD and DF carried out the experimental work and drafted the manuscript; QL, YM, XL, and SY analyzed the data and revised the manuscript; NL, HC, and HL designed the experiments and revised the manuscript. All authors read and approved the final manuscript.

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Correspondence to Hui Lin, Na Li or Hongge Chen.

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Dong, S., Fan, D., Liu, Q. et al. Enantioselectivity and key residue of Herbaspirillum huttiense monooxygenase in asymmetric epoxidation of styrenes. Appl Microbiol Biotechnol 106, 2007–2015 (2022). https://doi.org/10.1007/s00253-022-11843-z

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