益生芽孢杆菌脂肽类抑菌成分的高效液相色谱-电喷雾串联质谱分析

发布时间：2018-11-01 12:21

【摘要】：本文旨在探明一株具有益生潜质的枯草芽孢杆菌N2-10对肠道菌的抗菌作用及其活性成分,为深入评价该菌株的益生功能和益生机理奠定基础。以常见的肠道致病菌和有益菌为指示菌,采用抑菌圈法对该菌株发酵液成分的抑菌活性进行测定,并通过高效液相色谱-电喷雾串联质谱(HPLC-ESI-MS/MS)对其中的活性成分进行分析。结果表明:枯草芽孢杆菌N2-10发酵液的氯仿和正丁醇提取液对供试菌无明显的抑制作用;脂肽类对大肠杆菌、痢疾杆菌和沙门氏菌等肠道致病菌表现出明显的抑菌作用,而对肠道有益菌保加利亚乳杆菌抑制作用很弱,对嗜热链球菌则无抑制作用。脂肽粗提物的HPLC-ESI-MS/MS分析结果显示,脂肽粗提物中的脂肽类抗生素为伊枯草素家族中的C14~C17抗霉枯草菌素(Mycosubtilin)的同系物。因此推断,枯草芽孢杆菌N_2-10对肠道致病菌产生抑制作用依赖于其产生的脂肽抗生素Mycosubtilin。
[Abstract]:The purpose of this paper is to find out the antibacterial effect and active components of a probiotic Bacillus subtilis N2-10 against intestinal bacteria, and to lay a foundation for further evaluation of probiotic function and probiotic mechanism of Bacillus subtilis N2-10. The antimicrobial activity of the components of the fermentation broth of this strain was determined by the bacteriostasis circle method with the common enteric pathogenic bacteria and beneficial bacteria as the indicator bacteria. The active components were analyzed by high performance liquid chromatography-electrospray tandem mass spectrometry (HPLC-ESI-MS/MS). The results showed that chloroform and n-butanol extract from fermentation broth of Bacillus subtilis N2-10 had no obvious inhibitory effect on the tested bacteria. Lipopeptide showed obvious bacteriostatic effect on intestinal pathogenic bacteria such as Escherichia coli, Shigella dysenteriae and Salmonella, but had little inhibitory effect on Lactobacillus bulgaricus, but not on Streptococcus thermophilus. The HPLC-ESI-MS/MS analysis of lipopeptide crude extract showed that the lipopeptide antibiotics in the crude lipopeptide extract were the congeners of C14~C17 anti-mycomycin (Mycosubtilin) in the eucosin family. Therefore, it is inferred that the inhibitory effect of Bacillus subtilis NSP 2-10 on the production of intestinal pathogenic bacteria depends on the lipopeptide antibiotic Mycosubtilin. produced by it.
【作者单位】：河北农业大学生命科学学院;
【基金】：河北省科技支撑计划项目(12226605) 河北省高等学校科学技术研究项目(2011232)
【分类号】：S816

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