化学
酒
有机化学
光学(聚焦)
绿色化学
生物催化
组合化学
催化作用
反应机理
物理
光学
作者
Fulong Li,Yuwen Wei,Yan Du,Youxiang Liang,Yukun Zheng,Songhua Yuan,Huimin Yu
标识
DOI:10.1002/cctc.202301474
摘要
Abstract Alcohol dehydrogenases (ADHs) have garnered recognition for their potential in the synthesis of pivotal pharmaceutical compounds. However, their utilization in the context of piperidone synthesis remains an area ripe for exploration. In this study, we examine the performance of an alcohol dehydrogenase derived from Corynebacterium glutamicum ( Cg ADH) using a substrate analogue functional screening (SAFS) method to elucidate its substrate specificity. To improve the catalytic activity of Cg ADH, a phenylalanine/alanine‐scanning and iterative saturation mutation (PAS‐ISM) method was used. The most active variant, I151F/I195A, exhibited a remarkable 10.6‐fold increase in catalytic activity compared to the wild‐type. Structural analysis revealed that the introduction of residues 151F and 195A led to a remodeling of the substrate‐binding pocket, enabling additional p‐π hydrophobic interactions with the substrate, ultimately promoting a more favorable substrate binding pose. This study introduces the SAFS screening method, which enables the identification of enzymes with no sequence homology to known enzymes. Furthermore, the application of PAS‐ISM presents an efficient approach for the engineering of alcohol dehydrogenases. These findings open up promising avenues to expand the utility of ADHs in the synthesis of piperidone, thereby advancing the field of pharmaceutical chemistry.
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