Key Laboratory of Environmental and Applied Microbiology, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu 610041, P.R. China.
Schoolof Bioengineering, Sichuan University of Science and Engineering, Zigong 643000, P.R. China.
J Microbiol Biotechnol. 2019 Nov 28;29(11):1769-1776. doi: 10.4014/jmb.1805.04058.
Ethyl ()-3-hydroxy-3-(2-thienyl)propanoate(()-HEES)acts as a key chiral intermediate for the blockbuster antidepressant drug duloxetine, which canbe achieved viathe stereoselective bioreduction ofethyl 3-oxo-3-(2-thienyl) propanoate (KEES) that containsa 3-oxoacyl structure.The sequences of the short-chain dehydrogenase/reductases from sp. CA49 were analyzed, and the putative3-oxoacyl-acyl-carrier-protein reductase, KRED12, was able to stereoselectivelycatalyze theNADPH-dependent reduction to produce ()-HEES.The reductase activity of KRED12 towardsothersubstrates with 3-oxoacyl structure were confirmed with excellent stereoselectivity (>99% enantiomeric excess) in most cases. When coupled with a cofactor recycling system using glucose dehydrogenase, the KRED12 was able to catalyze the complete conversion of 100 g/l KEES within 12h, yielding the enantiopure product with >99% ee, showing a remarkable potential to produce ()-HEES.
（）-3-羟基-3-(2-噻吩基）丙酸乙酯（）-HEES 是一种关键的手性中间体，用于制造畅销的抗抑郁药度洛西汀，可以通过含有 3-氧代酰基结构的乙基 3-氧代-3-(2-噻吩基）丙酸酯（KEES）的立体选择性生物还原来实现。对 sp. CA49 的短链脱氢酶/还原酶序列进行了分析，假定的 3-氧代酰基-酰基-载体蛋白还原酶 KRED12 能够立体选择性地催化 NADPH 依赖性还原，生成（）-HEES。在大多数情况下，KRED12 对具有 3-氧代酰基结构的其他底物的还原酶活性表现出极好的立体选择性（>99%对映体过量）。当与使用葡萄糖脱氢酶的辅酶再生系统偶联时，KRED12 能够在 12 小时内催化 100g/L KEES 的完全转化，生成>99%ee 的对映体纯产物，显示出生产（）-HEES 的显著潜力。
