棒状链霉菌中隐性羊毛硫肽CLA 124的半体外生物合成  

Semi-in vitro biosynthesis of cryptic lanthipeptide CLA 124 from Streptomyces clavuligerus

作　　者：张铮[1,2] 张丽[1,2] 张杰[1,2] 马宏初 孙树涛[1] 钟瑾[1] 

机构地区：[1]中国科学院微生物研究所,北京100101 [2]中国科学院大学,北京100049

出　　处：《微生物学报》2015年第11期1402-1408,共7页Acta Microbiologica Sinica

基　　金：中国科学院知识创新重要方向项目(KSCX2-EW-Q-14;KSCX2-EW-J-6)~~

摘　　要：【目的】利用半体外生物合成方法获得棒状链霉菌中的隐性羊毛硫肽,为放线菌中羊毛硫肽资源的挖掘提供借鉴。【方法】利用nisin修饰系统,在大肠杆菌(Escherichia coli)中对隐性羊毛硫肽的核心肽进行体内修饰。修饰后产物经亲和层析和高效液相色谱(HPLC)纯化后,体外酶切反应切除前导肽,利用MALDITOF MS检测核心肽的脱水情况,并结合二级质谱解析其成环结构。【结果】获得了新的羊毛硫肽CLA 124,其核心肽脱去了4分子水,并形成2个硫醚键和一个二硫键。【结论】半体外生物合成方法可用于放线菌来源隐性羊毛硫肽的资源挖掘。[ Objective ] To obtain the cryptic lanthipeptide from Streptomyces clavuligerus by semi-in vitro biosynthesis that is a novel method for mining lanthipeptides resource from Streptomyces. [ Methods ] The core peptide of cryptic lanthipeptide was modified in E. coli by nisin modification system, and purified by affinity chromatography and High Performance Liquid Chromatography （HPLC）. After the leader peptide was removed, the core peptide was obtained and its dehydration and cyclic structure were analyzed by MALDI-TOF MS and tandem MS. [ Results] A novel lanthipeptide named CLA 124 with 4-fold dehydration 2 thioether bridges and one disulfide bridge was produced. [ Conclusion] Cryptic lanthipeptides from Streptomyces could be produced by semi-in vitro biosynthesis.

关 键 词：隐性基因簇 羊毛硫肽 半体外合成 脱水 环化 

分 类 号：Q936[生物学—微生物学]