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目的对1株海洋来源的菌株SCSIO 1682进行16SrDNA分子生物学鉴定,对发酵产物中的抑菌活性化合物进行分离鉴定,并对抑菌活性化合物的生物合成基因簇进行分析。方法通过构建基于16SrDNA序列的系统发育树来进行菌株鉴定;通过有机溶剂萃取、正相和反相硅胶层析等化学分离手段对菌株SCSIO1682的次级代谢产物进行活性追踪分离纯化;运用HR-ESI-MS,1 H和13 C NMR等波谱手段对所得化合物进行结构解析;通过对菌株SCSIO 1682进行全基因组扫描测序鉴定所得化合物的生物合成基因簇。结果该菌株被鉴定为链霉菌,命名为Streptomyces sp.SCSIO 1682,所得抑菌活性化合物为那西肽(nosihetide),鉴定了那西肽的生物合成基因簇并进行生物信息学分析。结论从海洋链霉菌Streptomyces sp.SCSIO 1682中分离鉴定了那西肽,那西肽生物合成基因簇的鉴定为利用组合生物合成和合成生物学技术对那西肽进行结构改造提供了新的基因资源。
Objective To identify 16S rDNA of a strain of marine origin SCSIO 1682, isolate and identify the antibacterial compounds in the fermentation product, and analyze the biosynthetic gene cluster of antibacterial compounds. Methods The phylogenetic tree based on 16S rDNA sequence was constructed to identify the strains. The secondary metabolites of strain SCSIO1682 were isolated and purified by organic solvent extraction, normal phase and reverse phase silica gel chromatography. MS, 1 H and 13 C NMR. The biosynthetic gene cluster of the obtained compounds was identified by genome-wide sequencing of strain SCSIO 1682. Results The strain was identified as Streptomyces sp. SCSIO 1682, and the resulting antibacterial activity was nosihetide. The biosynthetic gene cluster of nosiheptide was identified and bioinformatics analysis was performed. Conclusion The nosiheptide was isolated and identified from Streptomyces sp. SCSIO 1682, and the identification of nosiheptide biosynthetic gene cluster provided a new genetic resource for the structural transformation of nosiheptide using combinatorial biosynthesis and synthetic biology techniques .