一株粘细菌Myxococcus sp. STXZ90分离鉴定及生物活性分析.docVIP

一株粘细菌Myxococcus sp. STXZ90的分离鉴定及生物活性分析 　　DOI：10.7612/j.issn2017.02.004 　　摘要采用灭活大肠杆菌诱导法分离粘细菌，通过形态观察、生理生化实验、16S rRNA基因序列同源性分析，对其进行鉴定并归类.并通过平板对峙法、肿瘤细胞毒性实验等检测粘细菌代谢产物的抑菌和抗肿瘤活性，RPHPLC分离抗肿瘤活性物质.分离得到一株粘细菌，鉴定结果表明该菌株为黄色粘球菌，将其命名为Myxococcus sp. STXZ90（简写为STXZ90）.抑菌试验表明，STXZ90对多种人类病原菌及鱼类致病细菌如产气肠杆菌、嗜水气单胞菌等具有较高的抑制活性；STXZ90发酵上清的甲醇粗提物对多种肿瘤细胞具有广谱高效的抑制肿瘤细胞活性，对Hep3B，Hela和HUVEC等的最低半抑制浓度（IC50）为0.279～1.228 mg/L.对该甲醇粗提物进行RPHPLC分离，得到2个较纯的单峰物质，体外肿瘤细胞实验结果显示该组分有较强的抑制肿瘤细胞活性. 　　关键词粘细菌；次级代谢产物；高效液相色谱 　　中图分类号Q939文献标识码A文章编2017 　　On Isolation， Identification and Bioactivitiy of Myxococcus sp. STXZ90 　　HUANG Tonglong， LEI Lianghuan， LIAO Jiyan， ZHOU Cai， WU Yujing， 　　LI Chun， WEI Hui， XIA Liqiu， DING Xuezhi， ZHANG Youming* 　　（State Key Laboratory Breeding Base of Microbial Molecular Biology， College of Life Science， Hunan Normal University， Changsha 410081， China） 　　AbstractThe inactivated Escherichia. colidependent fruiting baiting technique was used to isolate myxobacterial strains. Based on the phylogenetical characteristics， physiological and biochemical properties and 16S rRNA gene sequence homology analysis， the isolated strain has been identified and classified. The inhabitation of the strain to different pathogens and tumor cells were detected by the Plate Confrontation Culture Method and tumor cell toxicity experiment. The antitumor substance was further purified by RPHPLC. Our results showed that the strain belonged to myxococcus xanthus and thus it was named as Myxococcus sp. STXZ90（STXZ90）. The antibacterial experiment indicated that STXZ90 had bacteriostatic activity to different human pathogens and fish pathogens， such as Enterobacter aerogenes， and Aeromonas hydrophil. The methanol elution of the fermented supernatant of STXZ90 exhibited a broad spectrum and high effect on various tumor cell lines. The IC50（half maximal inhibitory concentration） to Hep3B，Hela， and HUVEC were between 0.279 to 1.228 mg/L. The methanol elution was further purified by