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

作者：Tang, Jiawei; Wei, Tengyun; Ni, Guowei; Gu, Xiang; Wu, Yuanjie; Zhang, Fuli^*; Chen, Shaoxin^*

来源：Applied Biochemistry and Biotechnology, 2019, 188(1): 87-100.

DOI：10.1007/s12010-018-2904-2

• (R)-[3
• 5-bis(trifluoromethyl)phenyl] ethanol
• 3
• 5-bis(trifluoromethyl) acetophenone
• Carbonyl reductase
• Leifsonia sp
• Response surface methodology

(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 600g/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 59g/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.

• 出版日期2019-5
• 单位上海医药工业研究院