The activation of carboxylic acids to thioesters plays an important role in biology. However, biochemical studies and biotechnological applications are hampered by a general lack of access to thioesters, especially those based on Coenzyme A (CoA-SH). Here we show a generic thioester recycling enzyme by exploiting the promiscuous activity of a carboxylic acid reductase (CARsr). The adenylation domain of CARsr (CARsr-A) catalyses the conversion of a wide range of carboxylic acids to acyl-S-Coenzyme A and other thioesters in good yields. CARsr-A was used in situ as part of a recycling system to regenerate thioesters for acyl-S-Coenzyme A-dependent enzymes in one-pot reactions. This concept of thioester recycling is demonstrated with a range of acyltransferases that allow the formation of diverse amides and the non-native acylation of lysine side chains in a histone-derived peptide using the epigenetic writer, lysine acetyltransferase HATp300. Overall, these results establish a generic platform for thioester formation towards amide formation and beyond.

羧酸活化为硫酯在生物学中起着重要作用。然而，生化研究和生物技术应用受到普遍缺乏硫酯的阻碍，尤其是那些基于辅酶 A (CoA-SH) 的硫酯。在这里，我们通过利用羧酸还原酶 (CAR sr ) 的混杂活性展示了一种通用的硫酯回收酶。CAR sr (CAR sr -A)的腺苷酸化结构域催化各种羧酸以良好的产率转化为酰基-S-辅酶 A 和其他硫酯。汽车_-A 原位用作回收系统的一部分，以在一锅反应中为酰基-S-辅酶 A 依赖性酶再生硫酯。这种硫酯循环的概念通过一系列酰基转移酶得到证明，这些酰基转移酶允许使用表观遗传作者赖氨酸乙酰转移酶 HATp300 形成不同的酰胺和组蛋白衍生肽中赖氨酸侧链的非天然酰化。总的来说，这些结果为硫酯形成向酰胺形成及以后的形成建立了一个通用平台。