Stereoselective bioreduction of ethyl 3-oxo-3-(2-thienyl) propanoate using the short-chain dehydrogenase/reductase ChKRED12.
Stereoselective bioreduction of ethyl 3-oxo-3-(2-thienyl) propanoate using the short-chain dehydrogenase/reductase ChKRED12.
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DOI：
10.4014/jmb.1805.04058
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发表时间：
2019-11
影响因子：
2.8
通讯作者：
Zhi-qiang Ren;Yan Liu;Xiaoqiong Pei;Z. Wu
中科院分区：
文献类型：
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作者：
Zhi-qiang Ren;Yan Liu;Xiaoqiong Pei;Z. Wu
(S)-3-羟基-3-(2-噻吩基)丙酸乙酯((S)-HEES)是重磅抗抑郁药物度洛西汀的关键手性中间体，它可通过对含有3-氧代酰基结构的3-氧代-3-(2-噻吩基)丙酸乙酯(KEES)进行立体选择性生物还原而得到。对金黄杆菌属(Chryseobacterium sp.)CA49的短链脱氢酶/还原酶的序列进行了分析，推定的3-氧代酰基 - 酰基 - 载体 - 蛋白还原酶ChKRED12能够立体选择性地催化依赖NADPH的还原反应以生成(S)-HEES。在大多数情况下，证实了ChKRED12对具有3-氧代酰基结构的其他底物具有还原酶活性，且具有优异的立体选择性(>99%ee)。当与使用葡萄糖脱氢酶的辅因子循环系统偶联时，ChKRED12能够在12小时内催化100 g/L的KEES完全转化，生成对映体纯产物，ee值>99%，显示出生产(S)-HEES的显著潜力。
Ethyl (S)-3-hydroxy-3-(2-thienyl)propanoate((S)-HEES)acts as a key chiral intermediate for the blockbuster antidepressant drug duloxetine, which can be achieved via the stereoselective bioreduction of ethyl 3-oxo-3-(2-thienyl) propanoate (KEES) that contains a 3-oxoacyl structure. The sequences of the short-chain dehydrogenase/reductases from Chryseobacterium sp. CA49 were analyzed, and the putative3-oxoacyl-acyl-carrier-protein reductase, ChKRED12, was able to stereoselectively catalyze the NADPH-dependent reduction to produce (S)-HEES. The reductase activityof ChKRED12 towards other substrates with 3-oxoacyl structure were confirmed with excellent stereoselectivity (>99%ee) in most cases. When coupled with a cofactor recycling system using glucose dehydrogenase, the ChKRED12 was able to catalyze the complete conversion of 100 g/LKEES within 12 h, yielding the enantiopure product with >99% ee, showing a remarkable potential to produce(S)-HEES.