Department of Chemistry, Princeton University, Princeton, New Jersey 08544, United States.
Department of Chemistry, Johns Hopkins University, Baltimore, Maryland 21218, United States.
ACS Chem Biol. 2023 Aug 18;18(8):1854-1862. doi: 10.1021/acschembio.3c00281. Epub 2023 Jul 18.
Enediyne antibiotics are a striking family of DNA-cleaving natural products with high degrees of cytotoxicity and structural complexity. Microbial genome sequences, which have recently accumulated, point to an untapped trove of "cryptic" enediynes. Most of the cognate biosynthetic gene clusters (BGCs) are sparingly expressed under standard growth conditions, making it difficult to characterize their products. Herein, we report a fluorescence-based DNA cleavage assay coupled with high-throughput elicitor screening for the rapid, targeted discovery of cryptic enediyne metabolites. We applied the approach to , which harbors two such BGCs with unknown products, identified steroids as effective elicitors, and characterized 10 cryptic enediyne-derived natural products, termed clavulynes A-J with unusual carbonate and terminal olefin functionalities, with one of these congeners matching the recently reported jejucarboside. Our results contribute to the growing repertoire of enediynes and provide a blueprint for identifying additional ones in the future.
烯二炔类抗生素是一类具有高度细胞毒性和结构复杂性的 DNA 切割天然产物，引人注目。最近积累的微生物基因组序列表明，存在大量未被开发的“隐匿”烯二炔类抗生素。大多数同源生物合成基因簇（BGCs）在标准生长条件下表达很少，因此难以表征其产物。本文报道了一种基于荧光的 DNA 切割测定法，并结合高通量诱导剂筛选，用于快速、有针对性地发现隐匿烯二炔类代谢产物。我们将该方法应用于含有两个未知产物 BGCs 的 ，鉴定出甾体为有效诱导剂，并对 10 种隐匿烯二炔类衍生的天然产物进行了表征，这些天然产物被命名为clavulynes A-J，具有不寻常的碳酸盐和末端烯烃官能团，其中一种同化物与最近报道的 jejucarboside 相匹配。我们的研究结果丰富了烯二炔类抗生素的种类，并为未来识别更多的烯二炔类抗生素提供了蓝图。
