自1932年从巴氏酵母中首次发现旧的黄色酶1(OYE1)以来,OYEs对活化烯烃的生物催化不对称还原已成为有机合成中有价值的反应。为了获得立体互补的C=C键生物还原,已经进行了新OYEs的挖掘,特别是现有OYEs的蛋白质工程,在几种情况下成功实现了立体互补还原,进一步提高了应用潜力。在这次审查中,我们分析了结构,活跃的网站,和OYEs的底物识别,是其底物特异性和立体特异性的基础。还构建了OYEs超家族的序列相似性网络,以研究表征OYEs的范围。然后回顾并讨论了在过去十年(2009-2020年)中转换OYE的立体选择性并因此获得立体互补生物还原的结构指导工程,这可能会为相关生物催化剂的开采和工程提供新的见解。关键点:•构建并注释OYEs超家族的序列相似性网络。•比较来自不同类别的OYE的结构和活性位点。•“左/右”结合模式用于解释OYE的立体参照。•审查了OYE的结构引导工程以转换其立体选择性。
Since the first discovery of old yellow enzyme 1 (OYE1) from Saccharomyces pastorianus in 1932, biocatalytic asymmetric reduction of activated alkenes by OYEs has become a valuable reaction in organic synthesis. To access stereocomplementary C=C-bond bioreduction, the mining of novel OYEs and especially the protein engineering of existing OYEs have been performed, which successfully achieved the stereocomplementary reduction in several cases and further raise the potential of applications. In this review, we analyzed the structures, active sites, and substrate recognition of OYEs, which are the bases for their substrate specificity and stereospecificity. Sequence similarity network of OYEs superfamily was also constructed to investigate the scope of characterized OYEs. The structure-guided engineering to switch the stereoselectivity of OYEs and thus access stereocomplementary bioreduction over the last decade (2009-2020) was then reviewed and discussed, which might give new insights into the mining and engineering of related biocatalysts. KEY POINTS: • The sequence similarity network of OYEs superfamily was constructed and annotated. • The structures and active sites of OYEs from different classes were compared. • \"Left/right\" binding mode was used to explain the stereopreferences of OYEs. • Structure-guided engineering of OYEs to switch their stereoselectivity was reviewed.