Expanding the toolbox of enzymatic reactions accessible to organic chemists is one of the major goals in biocatalysis. Here we describe the development of an acyltransferase variant from Mycobacterium smegmatis in which a strategic Ser/Cys exchange in the catalytic triad dramatically expanded its synthetic capability to yield a biocatalyst able to efficiently catalyse the formation of thioesters and tertiary amides in water. Preparative scale (250 mM) biotransformations were performed starting from different thiols and secondary amines with excellent yields and reactions times, using vinyl esters as acylating agents. The high substrate-to-catalyst ratio and the cofactor independence make this process a sustainable and cost-effective procedure that was successfully applied to the synthesis of acetyl coenzyme A as well as structurally simpler analogues. Computational studies provided insights into the enzymatic selectivity and substrate recognition.

扩大有机化学家可利用的酶促反应工具箱是生物催化的主要目标之一。在这里，我们描述了耻垢分枝杆菌的酰基转移酶变体的发展其中催化三联体中的战略性Ser / Cys交换极大地扩展了其合成能力，从而生产出能够有效催化水中硫酯和叔酰胺形成的生物催化剂。使用乙烯基酯作为酰化剂，从不同的硫醇和仲胺开始，进行了制备规模（250 mM）的生物转化，转化率极高。高的底物与催化剂的比例和辅因子的独立性使该方法成为一种可持续且具有成本效益的方法，已成功地应用于乙酰辅酶A以及结构简单的类似物的合成。计算研究提供了对酶选择性和底物识别的见解。