Porterfield William B, Poenateetai Nannalin, Zhang Wenjun
Department of Chemical and Biomolecular Engineering, University of California, Berkeley, CA 94709, USA.
Department of Chemical and Biomolecular Engineering, University of California, Berkeley, CA 94709, USA; Chan Zuckerberg Biohub, San Francisco, CA 94158, USA.
iScience. 2020 Mar 27;23(3):100938. doi: 10.1016/j.isci.2020.100938. Epub 2020 Feb 26.
Polyketides produced by modular polyketide synthases (PKSs) are important small molecules widely used as drugs, pesticides, and biological probes. Tagging these polyketides with a clickable functionality enables the visualization, diversification, and mode of action study through bio-orthogonal chemistry. We report the de novo biosynthesis of alkyne-tagged polyketides by modular type I PKSs through starter unit engineering. Specifically, we use JamABC, a terminal alkyne biosynthetic machinery from the jamaicamide B biosynthetic pathway, in combination with representative modular PKSs. We demonstrate that JamABC works as a trans loading system for engineered type I PKSs to produce alkyne-tagged polyketides. In addition, the production efficiency can be improved by enhancing the interactions between the carrier protein (JamC) and PKSs using docking domains and site-directed mutagenesis of JamC. This work thus provides engineering guidelines and strategies that are applicable to additional modular type I PKSs to produce targeted alkyne-tagged metabolites for chemical and biological applications.
由模块化聚酮合酶（PKSs）产生的聚酮化合物是重要的小分子，广泛用作药物、农药和生物探针。用可点击的官能团标记这些聚酮化合物能够通过生物正交化学实现可视化、多样化以及作用方式研究。我们报道了通过起始单元工程，由模块化I型PKSs从头生物合成炔烃标记的聚酮化合物。具体而言，我们将来自牙买加酰胺B生物合成途径的末端炔烃生物合成机制JamABC与代表性的模块化PKSs结合使用。我们证明JamABC作为一种反式加载系统，用于工程化I型PKSs以产生炔烃标记的聚酮化合物。此外，通过使用对接结构域和对JamC进行定点诱变来增强载体蛋白（JamC）与PKSs之间的相互作用，可以提高生产效率。因此，这项工作提供了工程指导方针和策略，适用于其他模块化I型PKSs，以生产用于化学和生物学应用的靶向炔烃标记代谢物。
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