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Regulation of P450 TleB catalytic flow for the synthesis of sulfur-containing indole alkaloids by substrate structure-directed strategy and protein engineering

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Abstract

Indole alkaloids have attracted considerable attention from synthetic chemists and biochemists for their structural diversity and important biological activities. Compared with traditional organic synthesis methods, the strategy of using cytochrome P450s’ extraordinary abilities to selectively activate carbon-hydrogen bonds to assist in the synthesis of various indole alkaloids has the characteristics of short synthetic route, mild conditions and high atomic economy. Here, we utilized P450 monooxygenases HinD and TleB to synthesize a novel 6/5/8 tricyclic product from (S)-N-((S)-1-(4-fluoro-1H-indol-3-yl)-3-hydroxypropan-2-yl)-2-mercapto-3-methylbutanamide through the substrate structure-directed strategy. TleB was more effective in catalyzing C–S coupling, and was used to synthesize a series of 6/5/8 tricyclic indololactam derivatives to provide drug candidates. Interestingly, the S–S coupling product was observed in HinD catalysis, which was a minor product in the wild-type TleB catalysis. With the help of protein engineering, we accurately regulated the catalytic flow and reversed the selectivity of TleB to obtain the S–S coupling product. At the same time, the reaction mechanism was reasonably speculated by means of site blocking and protein-substrate complex analysis.

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Acknowledgements

This work was supported by the National Key Research and Development Program of China (2018YFA0903200 to H.S.), Wuhan University, Undergraduate Training Programs for Innovation and Entrepreneurship of Wuhan University to Y.L., the Science and Technology Commission of Shanghai, and the National Natural Science Foundation of China (31900033, 21ZR1433700). The authors thank staff from BL19U1 beamline at Shanghai Synchrotron Radiation Facility (SSRF), China, for their assistance in X-ray crystal data collection.

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  1. These authors contributed equally to this work.

Authors and Affiliations

  1. College of Chemistry and Molecular Sciences, Wuhan University, Wuhan, 430072, China

    Xinying Ge, Yan Long, Bo Gu, Zixuan Yang, Yinyin Feng, Shuo Zheng, Yingying Li & Heng Song

  2. School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, 200240, China

    Jun Wang & Wupeng Yan

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  1. Xinying Ge
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  2. Yan Long
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  3. Jun Wang
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  7. Shuo Zheng
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Correspondence to Wupeng Yan or Heng Song.

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11426_2023_1755_MOESM1_ESM.docx

Regulation of P450 TleB Catalytic Flow for the Synthesis of Sulfur-Containing Indole Alkaloids by Substrate Structure-Directed Strategy and Protein Engineering, approximately 27.9 MB.

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Ge, X., Long, Y., Wang, J. et al. Regulation of P450 TleB catalytic flow for the synthesis of sulfur-containing indole alkaloids by substrate structure-directed strategy and protein engineering. Sci. China Chem. 66, 3232–3241 (2023). https://doi.org/10.1007/s11426-023-1755-4

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  • DOI: https://doi.org/10.1007/s11426-023-1755-4

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