Leibniz Institute for Natural Product Research and Infection Biology, HKI, Beutenbergstrasse 11a, 07745, Jena, Germany.
Faculty of Biological Sciences, Friedrich Schiller University Jena, 07743, Jena, Germany.
Angew Chem Int Ed Engl. 2020 Nov 23;59(48):21535-21540. doi: 10.1002/anie.202009107. Epub 2020 Sep 21.
Mining the genome of the food-spoiling bacterium Burkholderia gladioli pv. cocovenenans revealed five nonribosomal peptide synthetase (NRPS) gene clusters, including an orphan gene locus (bol). Gene inactivation and metabolic profiling linked the bol gene cluster to novel bolaamphiphilic lipopeptides with antimycobacterial activity. A combination of chemical analysis and bioinformatics elucidated the structures of bolagladin A and B, lipocyclopeptides featuring an unusual dehydro-β-alanine enamide linker fused to an unprecedented tricarboxylic fatty acid tail. Through a series of targeted gene deletions, we proved the involvement of a designated citrate synthase (CS), priming ketosynthases III (KS III), a type II NRPS, including a novel desaturase for enamide formation, and a multimodular NRPS in generating the cyclopeptide. Network analyses revealed the evolutionary origin of the CS and identified cryptic CS/NRPS gene loci in various bacterial genomes.
从引起食物腐败的伯克霍尔德氏菌(Burkholderia gladioli pv. cocovenenans)基因组中挖掘到五个非核糖体肽合成酶（NRPS）基因簇，其中包括一个孤儿基因座（bol）。基因失活和代谢谱分析将 bol 基因簇与具有抗分枝杆菌活性的新型 bolaamphiphilic 脂肽联系起来。通过化学分析和生物信息学相结合，阐明了 bolagladin A 和 B 的结构，它们是具有不寻常的脱氢-β-丙氨酸烯酰胺连接的脂环肽，融合了前所未有的三羧酸脂肪酸尾巴。通过一系列有针对性的基因缺失，我们证明了一个指定的柠檬酸合酶（CS）、启动酮合酶 III（KS III）、包括用于烯酰胺形成的新型去饱和酶的 II 型 NRPS 以及一个多模块 NRPS 参与了环肽的生成。网络分析揭示了 CS 的进化起源，并在各种细菌基因组中鉴定出了隐藏的 CS/NRPS 基因座。
