Protein Engineering for Enhanced Acyltransferase Activity, Substrate Scope, and Selectivity of the Mycobacterium smegmatis Acyltransferase MsAcT,ACS Catalysis

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Protein Engineering for Enhanced Acyltransferase Activity, Substrate Scope, and Selectivity of the Mycobacterium smegmatis Acyltransferase MsAcT
ACS Catalysis ( IF 11.3 ) Pub Date : 2020-06-12 , DOI: 10.1021/acscatal.0c01767
Simon P. Godehard ₁ , Christoffel P. S. Badenhorst ₁ , Henrik Müller ₁ , Uwe T. Bornscheuer ₁

Affiliation

The highly efficient and versatile acyltransferase MsAcT from Mycobacterium smegmatis catalyzes aqueous acyl transfer reactions, enabling applications in environmentally friendly processes and enzyme cascades. We rationally designed several variants with up to 30-fold increased acyl transfer to hydrolysis ratios while mostly retaining initial activity. Variants exhibiting broader acyl-donor substrate scope and higher or inverted enantioselectivity were also designed. Alterations of the catalytic His-Asp pair decreased the activation of hydrolytic water, thereby increasing acyl transfer to hydrolysis ratios. This study demonstrates that targeting the disruption of water networks and manipulating the activation of nucleophiles are promising strategies for engineering promiscuous acyltransferase activities.

中文翻译：

用于增强耻垢分枝杆菌酰基转移酶MsAcT的酰基转移酶活性，底物范围和选择性的蛋白质工程

来自耻垢分枝杆菌的高效，多功能的酰基转移酶MsAcT催化水性酰基转移反应，可用于环保工艺和酶级联反应。我们合理地设计了几种变体，它们的酰基转移与水解比提高了30倍，同时大部分保留了初始活性。还设计了具有更宽泛的酰基供体底物范围和较高或反向对映选择性的变体。催化His-Asp对的改变降低了水解水的活化，从而增加了酰基转移与水解的比率。这项研究表明，针对水网络的破坏和操纵亲核试剂的活化是工程化混杂酰基转移酶活性的有前途的策略。

更新日期：2020-07-17

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