Probing the Acyl Carrier Protein-Enzyme Interactions within Terminal Alkyne Biosynthetic Machinery.
Probing the Acyl Carrier Protein-Enzyme Interactions within Terminal Alkyne Biosynthetic Machinery.
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探索末端炔生物合成机器中酰基载体蛋白-酶的相互作用。
DOI：
10.1002/aic.16355
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发表时间：
2018
期刊：
影响因子：
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通讯作者：
Zhang,Wenjun
中科院分区：
文献类型：
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作者：
Su,Michael;Zhu,Xuejun;Zhang,Wenjun
炔官能团由于其广泛的化学和生物应用而引起了人们的广泛兴趣。我们最近阐明了酰基载体蛋白（ACP）依赖的炔生物合成途径，然而，关于ACP与炔生物合成酶，酰基ACP连接酶（JamA）和膜结合双功能去饱和酶/乙炔酶（JamB）的相互作用知之甚少。在这里，我们表明JamB与ACP的相互作用比JamA更严格。此外，非同源ACP的定点突变显著改善了其与JamB的相容性，表明ACP-JamB界面可能存在静电相互作用。最后，易错PCR和第二个非同源ACP的筛选确定了ACP上对于与JamB相互作用重要的热点，并产生了被JamB更好识别的突变体。因此，我们的数据不仅提供了对炔生物合成中ACP相互作用的见解，而且还可能有助于未来炔标记代谢物的组合生物合成，用于化学和生物学应用。© 2018美国化学工程师学会AIChE J，64：4255-4262，2018
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 EngineersAIChE J, 64: 4255–4262, 2018