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Development of an Efficient and Cost-Effective Enzymatic Process for Production of (R)-[3,5-bis(trifluoromethyl)phenyl] Ethanol Using Carbonyl Reductase Derived from Leifsonia sp. S749

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Abstract

(R)-[3,5-bis(trifluoromethyl) phenyl] ethanol [(R)-3,5-BTPE] is a crucial chiral intermediate for the synthesis of the NK-1 receptor antagonists aprepitant, rolapitant and fosaprepitant. The carbonyl reductase KR01 from Leifsonia sp. S749, discovered by protein sequence alignment, could convert 3′,5′-bis(trifluoromethyl) acetophenone (3,5-BTAP) into (R)-3,5-BTPE with excellent activity and enantioselectivity. In order to enhance the conversion efficiency at high substrate concentrations, the reaction conditions were optimized by response surface analysis. The results showed that 600 g/L 3,5-BTAP was bioreduced to (R)-3,5-BTPE (> 99.9% enantiomeric excess) by the recombinant Escherichia coli/pET-28a (+)-KR01 whole cells, with a 98.3% conversion and 59 g/L/h productivity under the optimized reaction conditions. In addition, the recombinant E. coli cells could be repeatedly used up to seven times in the reaction mixture containing 90% isopropanol (IPA). This is the highest substrate loading and productivity for the bioreduction of 3,5-BTAP by carbonyl reductase ever reported, and this method represents an efficient and cost-effective process for production of (R)-3,5-BTPE.

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Contributors

Jiawei Tang is the first author. Shaoxin Chen, Fuli Zhang, and Jiawei Tang designed the study and wrote the paper. Jiawei Tang, Tengyun Wei, Guowei Ni, Xiang Guo, and Yuanjie Wu collected and analyzed the data. All authors have read and approved the final manuscript.

Funding

This work was supported by the National Major Scientific and Technological Special Project for “Significant New Drugs Development” (2014ZX09201001-005-001).

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Authors and Affiliations

  1. Shanghai Institute of Pharmaceutical Industry, China State Institute of Pharmaceutical Industry, 285 Gebaini Road, Pudong, Shanghai, 201203, China

    Jiawei Tang, Tengyun Wei, Guowei Ni, Xiang Guo, Yuanjie Wu, Fuli Zhang & Shaoxin Chen

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  1. Jiawei Tang
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  2. Tengyun Wei
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  3. Guowei Ni
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Correspondence to Fuli Zhang or Shaoxin Chen.

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Tang, J., Wei, T., Ni, G. et al. Development of an Efficient and Cost-Effective Enzymatic Process for Production of (R)-[3,5-bis(trifluoromethyl)phenyl] Ethanol Using Carbonyl Reductase Derived from Leifsonia sp. S749. Appl Biochem Biotechnol 188, 87–100 (2019). https://doi.org/10.1007/s12010-018-2904-2

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