Buyachuihan Lynn, Reiners Simon, Zhao Yue, Grininger Martin
Institute of Organic Chemistry and Chemical Biology, Buchmann Institute for Molecular Life Sciences, Goethe University Frankfurt, 60438, Frankfurt am Main, Germany.
Commun Chem. 2024 Aug 24;7(1):187. doi: 10.1038/s42004-024-01269-1.
Modular polyketide synthases (PKSs) play a vital role in the biosynthesis of complex natural products with pharmaceutically relevant properties. Their modular architecture makes them an attractive target for engineering to produce platform chemicals and drugs. In this study, we demonstrate that the promiscuous malonyl/acetyl-transferase domain (MAT) from murine fatty acid synthase serves as a highly versatile tool for the production of polyketide analogs. We evaluate the relevance of the MAT domain using three modular PKSs; the short trimodular venemycin synthase (VEMS), as well as modules of the PKSs deoxyerythronolide B synthase (DEBS) and pikromycin synthase (PIKS) responsible for the production of the antibiotic precursors erythromycin and pikromycin. To assess the performance of the MAT-swapped PKSs, we analyze the protein quality and run engineered polyketide syntheses in vitro. Our experiments include the chemoenzymatic synthesis of fluorinated macrolactones. Our study showcases MAT-based reprogramming of polyketide biosynthesis as a facile option for the regioselective editing of substituents decorating the polyketide scaffold.
模块化聚酮合酶（PKSs）在具有药学相关特性的复杂天然产物的生物合成中起着至关重要的作用。它们的模块化结构使其成为工程改造以生产平台化学品和药物的有吸引力的目标。在本研究中，我们证明了来自小鼠脂肪酸合酶的混杂丙二酰/乙酰转移酶结构域（MAT）可作为生产聚酮类似物的高度通用工具。我们使用三种模块化PKSs评估MAT结构域的相关性；短三模块的venemycin合酶（VEMS），以及负责生产抗生素前体红霉素和苦霉素的PKSs脱氧红霉内酯B合酶（DEBS）和苦霉素合酶（PIKS）的模块。为了评估MAT交换后的PKSs的性能，我们分析了蛋白质质量并在体外进行了工程化聚酮合成。我们的实验包括氟化大环内酯的化学酶促合成。我们的研究表明，基于MAT的聚酮生物合成重编程是对修饰聚酮支架的取代基进行区域选择性编辑的简便选择。
