东方拟无枝酸菌中ECO-0501生物合成基因簇的筛选及其相关调控基因的初步分析  

Screening ECO-0501 biosynthesis cluster from Amycolatopsis orientalis and preliminary analysis of related regulatory gene

作　　者：申洋[1] 黄鹤[2] 朱丽[3] 杨晟[2] 戈梅[3] 陈代杰[1] 

机构地区：[1]华东理工大学生物工程学院,上海200237 [2]中国科学院上海植物生理生态研究所,上海200032 [3]上海来益生物药物研究开发中心,上海201203

出　　处：《中国抗生素杂志》2013年第5期327-331,共5页Chinese Journal of Antibiotics

基　　金：国家自然科学基金(NFSC30672558);国家重大新药创制项目(2012ZX090301002-003)

摘　　要：ECO-0501是从东方拟无枝酸菌代谢物中分离到的一种新型线性多烯类化合物,其不但具有良好的抗菌活性,且体内毒性较低,目前处于临床前的开发。本文通过对东方拟无枝酸菌的菌落原位杂交,筛选到4个粘粒克隆NL5B,NL9-3-1,NL3-1C,NL3-3A,可以覆盖整个ECO-0501生物合成簇,并对其中的基因ORF4进行了生物信息学分析,推测其为ECO-0501合成途径中的正调控基因。并通过该基因的过表达,成功地将ECO-0501的产量提高了6倍,为下一步ECO-0501的开发及其调控机制的研究打下基础。ECO-0501 is a new linear polyene found from Amycolatopsis oriental＆. Because of the strong bacteridicial activity of ECO-0501 and its low toxity, preclinical development has been started on this antibiotic. Four positive cosmid clones were identified by colony hybridization that cover the whole ECO-0501 locus, including NL5B, NL9-3-1, NL3-1C and NL3-3A. Sequence analysis of the gene ORF4 revealed that its deduced protein might be a positive regulator controlling ECO-0501 biosynthesis. Overexpression of ORF4 showed that the yield of ECO- 0501 increased about six fold than that of wild-type strain. The present results provide us a tool for further study of ECO-0501 development and its regulatory mechanism.

关 键 词：ECO-0501 东方拟无枝酸菌 菌落原位杂交 调控蛋白 

分 类 号：Q784[生物学—分子生物学] TQ465.7[化学工程—制药化工]