链霉菌TJ430的鉴定及产物结构  

Isolation,identification of TJ430 strain and characterization of its antifungal metabolite

作　　者：杨少彬[1] 黄永春[1] 王常荣[1] 赵鹏[1] 张伟[1] 

机构地区：[1]农业部环境保护科研监测所,天津300191

出　　处：《微生物学报》2014年第6期624-634,共11页Acta Microbiologica Sinica

基　　金：天津市自然科学基金(12JCYBJC19900);中央级公益性科研院所基本科研业务费专项资金(农业部环境保护科研监测所)自主项目(2013SZJJHYC03)~~

摘　　要：【目的】通过对分离获得的土壤耐热放线菌株进行鉴定,并对该菌株产生的抗菌活性产物进行分离纯化和结构解析,达到判定该菌株归属及其产物新颖性目的。【方法】采用干热法对土壤样品进行前处理;采用经改良的HV培养基分离土壤耐热放线菌;通过琼脂块快速初筛法对分离获得的耐热放线菌进行生物活性快速评价;分别采用形态学观察法、细胞化学组分分析法、生理生化及酶学特征分析、16S rDNA序列分析、DNA杂交法,对分离获得的编号为TJ430的菌株进行菌株鉴定;采用柱层析法和制备色谱法,对发酵获得的有效成分粗提物进行分离纯化;采用红外光谱法、高分辨质谱法对分离获得的生物活性成分代谢产物进行结构探索。【结果】共分离获得570株耐热放线菌;抗菌活性快速初筛表明,编号为TJ430的菌株表现出优良的抗卵菌及广谱抗真菌活性;采用多种方法对菌株TJ430进行鉴定,结果表明该菌株为一株卡吾尔链霉菌;最终分离获得了纯度高达98%以上的有效成分纯品化合物;该有效成分的分子式为C40H66N3O11,分子量为765;分子中应含有亚胺基、甲基、亚甲基、羰基、共价双键、异丙基等化学基团。【结论】目前对该抗菌活性成分的详细化学结构研究尚有待深入,但该抗菌活性成分极有可能是一种新型化合物,具有较好的开发应用前景。[ Obiective] To screen new agro-antibiotics, rare actinomycetes were isolated by improved separation methods from soil samples and the chemical structure of the antifungal active product was elucidated. [ Methods] Dry heating method was used for soil samples pretreatment and the improved HV separation medium for rare actinomycetes separation; agar block rapid screening was used for the rapid evaluation of rare actinomycetes biological activity. For the identification of a strain numbered TJ430, morphology observation, cell chemical composition analysis, physiological and biochemical analysis, enzymology characteristics analysis, 16 S rDNA sequence analysis, and DNA hybridization method were used. Bioactive crude extract from fermentation was purified by column chromatography and preparative chromatography; infrared spectroscopy and high resolution mass spectrometry was used for structure elucidation of bioactive ingredient. [ Results] A total of 570 rare actinomycetes strains were isolated. Antibacterial activity of rapid screen showed that the numbed TJ430 strain showed excellent anti oomycetes and broad-spectrum antifungal activity. Strain identification resuhs show that the strain is a S. cavourensis. The molecular formulas of the effective ingredient is C40H66N3O11, molecular weight is 765. Amino, methyl, methylene, carbonyl, covalent bond, isopropyl and other chemical groups should contained in the molecular. [ Conclusion] The characterized antibacterial active ingredient has good development prospect.

关 键 词：菌株鉴定 农用抗生素 分离纯化 结构鉴定 

分 类 号：Q93[生物学—微生物学]