南京农业大学学报2025，Vol.48Issue(4)：947-959,13.DOI:10.7685/jnau.202404009
地衣芽胞杆菌JNBS001抗菌物质的鉴定及其抑菌机制研究
Identification of antimicrobial substances from Bacillus licheniformis JNBS001 and study on its antibacterial mechanism
摘要
Abstract
[Objectives]The aim of this paper was to identify the antimicrobial substances in the fermentation product of a strain of Bacillus licheniformis JNBS001 screened from the natural environment,as well as its antibacterial activity and mechanism against Escherichia coli and Listeria monocytogenes,and to preliminarily explore the prospect of its application as an antimicrobial substance in the food preservative system.[Methods]The whole genome sequencing of JNBS001 was subjected,the synthetic gene clusters of secondary metabolites were predicted,and non-targeted identification was performed by LC-MS/MS.Subsequently,the inhibitory activity and spectrum of JNBS001 fermentation product and the minimal inhibitory concentration(MIC)of the lyophilized powder of the fermentation product against E.coli and L.monocytogenes were determined.In addition,the inhibitory mechanism of JNBS001 fermentation product was analyzed by measuring K+and Ca2+leakage,alkaline phosphatase(AKP)activity,cell membrane permeability and cell morphology observation.[Results]Sequencing results showed that the JNBS001 genome was 3 914 347 bp in length with 46.59%GC content,encoding a total of 3 971 genes,and predicted the synthetic gene clusters of 13 secondary metabolites,among which 7 gene clusters were complete with 100%similarity.Combined with LC-MS/MS non-targeted identification data,macrolactin H and bacilysin were confirmed as the antimicrobial substances in the fermentation products of JNBS001.The fermentation products of JNBS001 effective against E.coli,Salmonella,Staphylococcus aureus,L.monocytogenes,Enterobacter sakazakii and Micrococcus luteus had bacterio static activity.The results of the analysis of its antibacterial mechanism showed that the fermentation products of JNBS001 changed the cell morphology of E.coli and L.monocytogenes,increased membrane permeability,and made a large amount of extravasation of the cell contents,thus causing bacterial death.[Conclusions]The antibacterial substances in the fermentation products of JNBS001 included macrolactin H and bacilysin,and its fermentation products had broad-spectrum antibacterial properties,and the inhibition was realized by destroying the integrity of the bacterial cell membrane and cell wall structure.关键词
地衣芽胞杆菌/全基因组测序/LC-MS/MS/抑菌机制Key words
Bacillus licheniformis/whole genome sequencing/LC-MS/MS/antibacterial mechanism分类
轻工纺织引用本文复制引用
马渝童,孟凡强,陆兆新..地衣芽胞杆菌JNBS001抗菌物质的鉴定及其抑菌机制研究[J].南京农业大学学报,2025,48(4):947-959,13.基金项目
国家自然科学基金项目(32072182) （32072182）
