Devine Andrew J, Parnell Alice E, Back Catherine R, Lees Nicholas R, Johns Samuel T, Zulkepli Ainul Z, Barringer Rob, Zorn Katja, Stach James E M, Crump Matthew P, Hayes Martin A, van der Kamp Marc W, Race Paul R, Willis Christine L
School of Chemistry University of Bristol BS81TS Bristol UK.
School of Biochemistry University of Bristol BS81TD Bristol UK.
Angew Chem Weinheim Bergstr Ger. 2023 Jan 16;135(3):e202213053. doi: 10.1002/ange.202213053. Epub 2022 Dec 8.
Abyssomicin C and its atropisomer are potent inhibitors of bacterial folate metabolism. They possess complex polycyclic structures, and their biosynthesis has been shown to involve several unusual enzymatic transformations. Using a combination of synthesis and in vitro assays we reveal that AbyV, a cytochrome P450 enzyme from the gene cluster, catalyses a key late-stage epoxidation required for the installation of the characteristic ether-bridged core of abyssomicin C. The X-ray crystal structure of AbyV has been determined, which in combination with molecular dynamics simulations provides a structural framework for our functional data. This work demonstrates the power of combining selective carbon-13 labelling with NMR spectroscopy as a sensitive tool to interrogate enzyme-catalysed reactions in vitro with no need for purification.
阿比西霉素C及其阻转异构体是细菌叶酸代谢的强效抑制剂。它们具有复杂的多环结构，并且其生物合成已显示涉及几种不寻常的酶促转化。通过合成与体外测定相结合的方法，我们发现来自该基因簇的细胞色素P450酶AbyV催化阿比西霉素C特征性醚桥核心形成所需的关键后期环氧化反应。已确定AbyV的X射线晶体结构，其与分子动力学模拟相结合为我们的功能数据提供了结构框架。这项工作证明了将选择性碳-13标记与核磁共振光谱相结合作为一种灵敏工具在体外研究酶催化反应而无需纯化的强大作用。
