Peña-Ortiz Luis, Graça Ana Patrícia, Guo Huijuan, Braga Daniel, Köllner Tobias G, Regestein Lars, Beemelmanns Christine, Lackner Gerald
Junior Research Group Synthetic Microbiology , Leibniz Institute for Natural Product Research and Infection Biology (HKI) , Beutenbergstr. 11a , 07745 Jena , Germany . Email:
Friedrich Schiller University , Beutenbergstr. 11a , 07745 Jena , Germany.
Chem Sci. 2020 Apr 23;11(20):5182-5190. doi: 10.1039/d0sc01172j. eCollection 2020 May 28.
Mycofactocin (MFT) is a redox cofactor belonging to the family of ribosomally synthesized and post-translationally modified peptides (RiPPs) and is involved in alcohol metabolism of mycobacteria including . A preliminary biosynthetic model had been established by bioinformatics and studies, while the structure of natural MFT and key biosynthetic steps remained elusive. Here, we report the discovery of glycosylated MFT by C-labeling metabolomics and establish a model of its biosynthesis in . Extensive structure elucidation including NMR revealed that MFT is decorated with up to nine β-1,4-linked glucose residues including 2--methylglucose. Dissection of biosynthetic genes demonstrated that the oligoglycosylation is catalyzed by the glycosyltransferase MftF. Furthermore, we confirm the redox cofactor function of glycosylated MFTs by activity-based metabolic profiling using the carveol dehydrogenase LimC and show that the MFT pool expands during cultivation on ethanol. Our results will guide future studies into the biochemical functions and physiological roles of MFT in bacteria.
真菌铁载体（MFT）是一种氧化还原辅因子，属于核糖体合成及翻译后修饰肽（RiPPs）家族，参与分枝杆菌的酒精代谢，包括……通过生物信息学和……研究已建立了初步的生物合成模型，而天然MFT的结构和关键生物合成步骤仍不清楚。在此，我们通过¹³C标记代谢组学报告了糖基化MFT的发现，并建立了其在……中的生物合成模型。包括核磁共振（NMR）在内的广泛结构解析表明，MFT最多被九个β-1,4-连接的葡萄糖残基修饰，包括2-甲基葡萄糖。对生物合成基因的剖析表明，寡糖基化由糖基转移酶MftF催化。此外，我们通过使用香芹醇脱氢酶LimC的基于活性的代谢谱分析证实了糖基化MFT的氧化还原辅因子功能，并表明在乙醇培养过程中MFT库会扩大。我们的结果将指导未来对MFT在细菌中的生化功能和生理作用的研究。
