The alkyne functionality has attracted much interest due to its diverse chemical and biological applications. We recently elucidated an acyl carrier protein (ACP)‐dependent alkyne biosynthetic pathway, however, little is known about ACP interactions with the alkyne biosynthetic enzymes, an acyl‐ACP ligase (JamA) and a membrane‐bound bifunctional desaturase/acetylenase (JamB). Here, we showed that JamB has a more stringent interaction with ACP than JamA. In addition, site‐directed mutagenesis of a non‐cognate ACP significantly improved its compatibility with JamB, suggesting a possible electrostatic interaction at the ACP‐JamB interface. Finally, error‐prone PCR and screening of a second non‐cognate ACP identified hot spots on the ACP that are important for interacting with JamB and yielded mutants which were better recognized by JamB. Our data thus not only provide insights into the ACP interactions in alkyne biosynthesis, but it also potentially aids in future combinatorial biosynthesis of alkyne‐tagged metabolites for chemical and biological applications. © 2018 American Institute of Chemical Engineers AIChE J, 64: 4255–4262, 2018

中文翻译：

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探索末端炔烃生物合成机制中的酰基载体蛋白-酶相互作用

炔烃的功能因其多样化的化学和生物学应用而引起了人们的极大兴趣。我们最近阐明了酰基载体蛋白（ACP）依赖的炔烃生物合成途径，但是，人们对ACP与炔烃生物合成酶，酰基-ACP连接酶（JamA）和膜结合双功能去饱和酶/乙酰化酶（JamB）的相互作用了解甚少。 。在这里，我们证明了JamB与ACP的互动比JamA更为严格。此外，非同源ACP的定点诱变显着提高了其与JamB的相容性，表明ACP-JamB界面可能存在静电相互作用。最后，容易出错的PCR和第二个非同源ACP的筛选确定了ACP上的热点，这些热点对于与JamB相互作用非常重要，并产生了可被JamB更好识别的突变体。因此，我们的数据不仅提供了对炔烃生物合成中ACP相互作用的见解，而且还潜在地有助于在将来的化学和生物学应用中炔烃标记的代谢物的组合生物合成。©2018美国化学工程师学会AIChE J，64：4255-4262，2018年