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Enantioselective synthesis of a chiral intermediate of himbacine analogs by Burkholderia cepacia lipase A

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Abstract

The enantiomers of (4R/S)-4-hydroxy-N, N-diphenyl-2-pentynamide are key chiral synthons for the synthesis of thrombin receptor antagonists such as vorapaxar. In this paper, we report the enzymatic preparation of enantiomerically enriched (4R)-4-hydroxy-N, N-diphenyl-2-pentynamide using lipase A from Burkholderia cepacia ATCC 25416 as the catalyst. First, the lipase gene (lipA) and its chaperone gene (lipB) was cloned and expressed in Escherichia coli system. After purification, lipase A activation was performed with the assistance of foldase lipase B. Enzyme assay revealed that the activated lipase A showed the optimal catalytic activity at 60 ºC and pH 7. The effects of various metals on the activity were investigated and results demonstrated that most of the metals inhibited the activity. To further improve the catalytic outcome, two-phase reaction was studied, and n-hexane proved to be a good organic solvent for the combination system. Using the optimize conditions, (4R)-4-hydroxy-N, N-diphenyl-2-pentynamide with 94.5% ee value and 48.93% conversion ratio was achieved. Our investigation on this lipase reveals lipase A as a promising biocatalyst for producing chiral propargyl alcohol for preparation of novel himbacine analogs.

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Acknowledgements

We express our appreciation to Huijiao Wen from Beijing University of Chemical Technology for insightful discussions and comments.

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  1. Authors Yanming Chen and Fei Gao have contributed equally to this work.

Authors and Affiliations

  1. Tianjin Key Laboratory for Modern Drug Delivery & High-Efficiency, School of Pharmaceutical Science and Technology, Tianjin University, 92 Weijin Road, Nankai District, Tianjin, 300072, People’s Republic of China

    Yanming Chen

  2. State Key Laboratory of Chemical Resources Engineering, Beijing University of Chemical Technology, Beijing, 100029, People’s Republic of China

    Yanming Chen, Fei Gao, Guojun Zheng & Shuaihua Gao

  3. Department of Chemistry, and California Institute for Quantitative Biosciences, University of California, Berkeley, 94720, USA

    Shuaihua Gao

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  1. Yanming Chen
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  2. Fei Gao
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  4. Shuaihua Gao
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Correspondence to Shuaihua Gao.

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Chen, Y., Gao, F., Zheng, G. et al. Enantioselective synthesis of a chiral intermediate of himbacine analogs by Burkholderia cepacia lipase A. Biotechnol Lett 42, 2643–2651 (2020). https://doi.org/10.1007/s10529-020-02969-z

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  • DOI: https://doi.org/10.1007/s10529-020-02969-z

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