目的以基因工程菌A.CJSl001与SIPI-GE.2006dorf20为研究对象,获得雷莫拉宁类似物同时测定其生物活性。方法利用产生菌的自然选育,雷莫拉宁类似物的分离纯化,HPLC检测产量变化及MIC检测。结果获得高产菌株A.CJSl011与SIPI-GE.2016dorf20,得到无糖雷莫拉宁的量为120mg无氯雷莫拉宁的量为203mg,其主要成分A2的HPLc显示纯度均大于90%。生物活性检测显示:无糖雷莫拉宁A,对耐甲氧西林金黄色葡萄球菌株ATCC43300的抑菌浓度为2μg/mL,对耐万古霉素肠球菌ATCC51299的抑菌浓度为1μg/mL;无氯雷莫拉宁A2对耐甲氧西林金黄色葡萄球菌株ATCC43300的抑菌浓度为21μg/mL,对耐万古霉素肠球菌株ATCC51299抑菌浓度为2gg/mL。结论以生物合成的方法分离纯化获得雷莫拉宁的类似物为进一步研究雷莫拉宁奠定了基础。
Objective To obtain ramoplanin analogues from the genetic engineered bacteria A.C JS1001 and SIPI-GE.2006 dorf20 and investigate their antimicrobial activities. Methods The methods of natural selection for genetic engineered bacteria; separation, purification, HPLC and MIC detection for ramoplanin analogues were used. Results 120mg ramoplanin aglycones and 203mg didechlororamoplanin(with 〉90% purity) were isolated from fermentation broth of the high yield strain A.C JS 1011 and SIPI-GE.2016 dorf20. Antimicrobial MICs of ramoplanin aglycones A2 to methicillin-resistant Staphylococcus aureus ATCC43300 and vancomycin resistant Enterococcus ATCC51299 are 2 and 1μg/mL. Both MICs of didechlororamoplanin A2 against Methicillin-resistant Staphylococcus aureus ATCC43300 MIC and vancomycin resistant Enterococcus ATCC51299 are 2μg/mL. Conclusion This biosynthesis method about the ramoplanin analogues has laid the foundation for further study.