醇脱氢酶
立体专一性
生物催化
活动站点
基质(水族馆)
酒
雷斯顿
化学
脱氢酶
立体化学
催化作用
组合化学
酶
生物化学
生物
反应机理
生态学
作者
Yingying Zhou,Yu Wang,Xi Chen,Jinhui Feng,Min Wang,Qingping Wu,Dunming Zhu
标识
DOI:10.1016/j.enzmictec.2021.109837
摘要
Enzymatic stereospecific reduction of 17-oxosteroids offers an attractive approach to access 17β-hydroxysteroids of pharmaceutical importance. In this study, by adjusting the flexibility of α6-helix at the substrate entrance of the alcohol dehydrogenase from Ralstonia sp. (RasADH), the catalytic activity toward the stereospecific 17β-reduction of androstenedione was improved without sacrifice of the enantioselectivity. Among the mutants, F205I and F205A exhibited up to 623- and 523-fold improvement in catalytic efficiency, respectively, towards a range of different 17-oxosteroids compared to the wild-type enzyme. The corresponding 17β-hydroxysteroids were prepared in optically pure form with high space-time productivity and isolated yields using F205I as the biocatalyst, indicating that these mutants are promising biocatalysts for this useful transformation. These results suggest that modulating the flexibility of the active site lid offers an effective approach to engineer alcohol dehydrogenase for accommodating bulky steroidal substrates.
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