基于定向PCR和UPLC-MS技术分析放线菌中的产物  

Studies on secondary metabolites of actinomycete based on PCR and UPLC-MS/MS analysis

作　　者：万国庆[1,2] 杨志钧[2,3] 饶敏[2,3] 陈代杰[3] 戈梅[2] 倪孟祥[1] 

机构地区：[1]中国药科大学生命科学与技术学院,南京210009 [2]上海来益生物药物研发中心,上海201203 [3]上海交通大学药学院,上海200240

出　　处：《中国抗生素杂志》2014年第1期24-27,31,共5页Chinese Journal of Antibiotics

基　　金：国家重大专项资助"创新微生物高效筛选与发现技术平台研究"(编号:2012ZX09301002-003)

摘　　要：目的从放线菌中发现聚酮类或大环内酯环类化合物。方法采用定向PCR方法从放线菌中筛选含Ⅰ型PKS基因,然后利用UPLC-MS技术分析该菌株发酵产物。结果从40株放线菌中获得了18株PCR阳性菌株,进一步的UPLC-MS/MS分析,从菌株HCCB11494发现了7个macrotetrolides类抗生素;该菌株的种属初步鉴定为小链霉菌。结论采用PKS I基因定向筛选结合UPLC-MS分析可从放线菌中筛选得到大环内酯化合物。Objective To discover polyketide and macrolide compounds from metabolites of actinomycetes. Methods Type I polyketidesynthase genes was cloned from genomes of actinomycetes by PCR technology, then metabolites of actinomycetes were analyzed by UPLC-MS/MS method. Results 18 positive strains were screened from 40 strains by PCR technology. Seven macrotetrolides were discovered from the strain HCCB11494 by UPLC- MS/MS analysis. And this strain was preliminarily identified as Streptomyces parvus according to gyrB sequences. Conclusion Macrolide compounds could be discovered from aetinomyeetes by PCR technology in combination with UPLC-MS analysis.

关 键 词：定向PCR UPLC-MS MS I型PKS Macrotetrolides GYRB 

分 类 号：R978.1[医药卫生—药品]