Discovery of bioactive secondary products from Streptomyces based on genome mining strategy
Objective To discover active natural products from Streptomyces using a genome mining strategy.Methods Initially,the whole genome sequence of Streptomyces malaysiensis DSM 41697 was obtained through sequencing.The antiSMASH tool was utilized for the preliminary prediction of gene clusters responsible for potential secondary metabolite biosynthesis in Streptomyces malaysiensis.Subsequently,through the use of Blast,2ndFind,and other online tools,the target gene clusters were analyzed and the gene boundaries were confirmed.Following this,various fermentation conditions were tested to preliminarily determine the production of the target compounds using HPLC and LC-MS.Finally,the target compounds were isolated,purified,and their structures elucidated.Results Two biosynthetic gene clusters from Streptomyces malaysiensis were identified as padanamide synthesis and benzastatin biosynthesis,respectively.Through the analysis of online tools Blast and 2ndFind,the boundaries of the two clusters were confirmed,and all genes within the clusters were annotated.Subsequently,after optimizing the fermentation medium,three padanamide members and one benzastatin derivative,malaymycin,were purified.Conclusion Guided by the genome mining strategy,it is feasible to discover potential natural product biosynthetic gene clusters from microbial genomes and subsequently determine the products of the target gene clusters through fermentation optimization.This strategy showcases an efficient method for the directed discovery of novel bacterial natural products and may offer new biosynthetic pathways for combinatorial biosynthesis and synthetic biology.
NETL
NSTL
万方数据
