链霉菌次级代谢调控通路的时序调控机制研究

Streptomyces is the main industrial producing-strain of microbial drugs, the stability and strength of its main regulatory pathway for secondary metabolic process determines the efficiency of microbial drug biosynthesis. In this project with the novel antibiotic daptomycin from Streptomyces roseosporus as a model, by resolving the cross-talk mechanism between the GBL signaling pathway and Pho two-component system (TCS)，and revealing the post-translational modification and degradation mechanism of key regulatory proteins, the kinetics of these regulatory pathways will be traced and analyzed. Through establishing the intensity kinetic curve, the Robust characteristics of GBL signal cascade regulatory pathway (stability and intensity) is elucidated, and then the temporal regulatory mechanism of the secondary metabolic pathway in Streptomyces will be revealed. . Our research on the temporal regulation of daptomycin biosynthesis will bring forward a new angle for secondary metabolic regulatory pathway from the previous research on the transcriptional regulation mechanism to the dynamic mechanism. It not only provides the basis of Streptomyces engineering for the higher yield and better quality of microbial drugs, but also produces important significance both on academic study and industrial applications.

链霉菌是微生物药物的主要生产菌，其主要次级代谢调控通路的稳定性、调控强度决定微生物药物的生物合成效率。本项目以产达托霉素的玫瑰孢链霉菌为模式研究对象，通过解析GBL信号级联调控通路与磷酸双组份信号通路的交互作用机制、关键调控蛋白翻译后修饰与降解机制，对调控通路关键蛋白进行全生命过程追踪分析；建立其对达托霉素次级代谢过程的调控强度变化的动力学曲线，阐析GBL信号调控通路的鲁棒性状（稳定性、调控强度），揭示GBL信号级联通路对次级代谢途径的时序调控机制。. 本研究从以往的基因转录调控水平转到蛋白水平，对整条调控通路动力学机制进行研究，使我们更加深刻理解链霉菌复杂的调控机制，为重构链霉菌调控途径提供理论基础，具有重要的基础研究意义和工业应用价值。

链霉菌是微生物药物的主要生产菌，其药物生物合成过程受到次级代谢调控网络的严谨控制，因此对达托霉素生物合成过程的时序调控机制解析，为生产菌优质高产改造提供依据，具有重要的理论研究和产业应用意义。 项目以达托霉素的生产菌玫瑰孢链霉菌为模式研究对象，解析了GBL信号级联调控通路与磷酸双组份信号通路的交叉互作，阐析基因组甲基化修饰和巴豆酰化修饰对达托霉素次级代谢调控机制，揭示了关键调控蛋白AtrA降解的分子机制，从而系统解析了达托霉素生物合成过程的时序调控机制。同时，基于调控机制利用合成生物学技术构建了高产菌，实现了产业化。项目发表SCI论文22篇、中文核心刊物2篇，其中第一标注篇10篇。授权国内外发明专利8件，其中南非发明专利2件、PCT专利3件，受理美国发明专利1件；以第一作者申报的“放线菌药物高效生物合成关键技术及其产业应用”，获中国石油与化学工业联合会技术发明奖一等奖。