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目的:研究游动放线菌I03A-00723发酵产物95-3的分离纯化与体外生物学活性。方法:发酵I03A-00723菌株,收集菌丝体,用丙酮浸泡、乙酸乙酯萃取,通过硅胶柱色谱、半制备HPLC进行纯化。采用微稀释法和高通量荧光检测法分别测定化合物95-3的抗菌活性和对PDF酶的抑制活性。结果:确定了化合物95-3的分离纯化条件,HPLC显示其纯度达100%。体外活性实验结果表明:化合物95-3对粪肠球菌、屎肠球菌的敏感菌株和万古霉素耐药菌株均有较好的抗菌活性,对耐万古霉素屎肠球菌ATCC700221的MIC达到了3μg.mL-1;化合物95-3对PDF酶有较好的抑制活性。结论:化合物95-3具有较好的抗VRE活性和PDF酶抑制活性,有深入研究价值。
Objective: To study the isolation, purification and in vitro biological activity of the 95-3 actinomycete Actinomycete I03A-00723. Methods: The strain I03A-00723 was fermented, the mycelium was collected, soaked in acetone, extracted with ethyl acetate and purified by silica gel column chromatography and semi-preparative HPLC. The antibacterial activity of compound 95-3 and its inhibitory activity on PDF enzyme were determined by micro-dilution method and high-throughput fluorescence detection. Results: The compound 95-3 was isolated and purified. The purity of compound 95-3 was 100%. The results of in vitro activity showed that compound 95-3 had good antibacterial activities against Enterococcus faecalis, Enterococcus faecium and vancomycin-resistant strains, and MIC of vancomycin-resistant Enterococcus faecium ATCC700221 reached 3μg .mL-1; Compound 95-3 has better inhibitory activity on PDF enzyme. Conclusion: Compound 95-3 has good anti-VRE activity and PDF enzyme inhibitory activity, and has further research value.