论文部分内容阅读
目的比较银杏内生真菌的抗菌活性,并对其抗菌活性成分进行分析。方法采用组织分离法从健康银杏叶、枝条、果实等组织中分离、纯化内生真菌;以大肠杆菌、金黄色葡萄球菌、枯草芽胞杆菌、酿酒酵母、黑曲霉和根霉为抑菌指示菌,采用滤纸片和平板孔阱扩散法,对40株银杏内生真菌发酵液抗菌活性进行初筛和复筛;利用硅胶柱色谱、凝胶Sephadex LH-20柱色谱、制备型HPLC、薄层色谱(TLC)和HPLC-MS等方法,对抗菌活性较高的草酸青霉菌Penicillium oxalicum YXY009菌株的抗菌成分进行分析。结果 40株银杏内生真菌中,18株为不产孢真菌;产孢真菌中,青霉属和曲霉属为优势菌属,分别占总菌数的17.5%和15.0%;YXY009菌株发酵液的抑菌活性明显高于其他菌株,对酿酒酵母和根霉抑菌圈直径分别达到(22.35±1.32)和(23.60±0.83)mm;从YXY009菌株发酵液中分离得到抑菌活性有效成分,初步鉴定为毛果杨苷。结论首次从银杏内生真菌代谢产物中分离出具有广谱抗菌活性的化合物毛果杨苷。
Objective To compare the antibacterial activity of endophytic fungi of Ginkgo biloba and analyze the antibacterial active ingredients. Methods Endophytic fungi were isolated and purified from healthy ginkgo biloba leaves, branches and fruits by tissue isolation method. Escherichia coli, Staphylococcus aureus, Bacillus subtilis, Saccharomyces cerevisiae, Aspergillus niger, The antibacterial activity of 40 strains of endophytic fungi fermentation broth was preliminarily screened and re-screened by filter paper and plate well-well diffusion method. The compounds were characterized by silica gel column chromatography, gel Sephadex LH-20 column chromatography, preparative HPLC and thin layer chromatography TLC) and HPLC-MS. The antibacterial components of Penicillium oxalicum YXY009 strain with higher antibacterial activities were analyzed. Results Among the 40 ginkgolic endophytic fungi, 18 were non-sporulating fungi. Penicillium and Aspergillus were the dominant genus in the spore fungi, accounting for 17.5% and 15.0% of the total number of bacteria, respectively. YXY009 fermentation broth The antibacterial activity was significantly higher than that of other strains, and the diameter of inhibition zone against Saccharomyces cerevisiae and Rhizopus were (22.35 ± 1.32) and (23.60 ± 0.83) mm, respectively. The antibacterial activity of YXY009 was isolated and identified As fruit poplar glycosides. Conclusion For the first time, the compound pilocarpine with broad-spectrum antibacterial activity was isolated from the metabolites of endophytic fungi from Ginkgo biloba.