赤红球菌二鸟苷酸环化酶介导的信号通路研究

红球菌在生物催化和转化、代谢工程和生物修复等领域得到了广泛的应用。红球菌属微生物如何感受外界化学物质并作出响应是一个重要的科学问题。本项目以筛选获得的有机溶剂耐受菌赤红球菌SD3为研究对象，在该菌中筛选二鸟苷酸环化酶互作蛋白和c-di-GMP效应蛋白，并进行验证。在此基础上，研究二鸟苷酸环化酶互作蛋白的配体结合结构域的作用，测定纳米磷脂盘中二鸟苷酸环化酶的活性，并对c-di-GMP效应蛋白进行生物信息学分析。此外，对这两种蛋白的基因分别进行敲除和回补，判断一些化学物质对基因敲除株和基因回补株的影响，从而进一步阐明这两种蛋白的功能。本项目能够深化人们关于赤红球菌二鸟苷酸环化酶介导的信号通路的认识，同时对于生物制备过程和环境污染物的生物修复等方面都具有重要的实践意义。

Rhodococcus has a wide application in the field of biocatalysis and biotransformation, metabolic engineering and bioremediation. How Rhodococcus senses and responses to chemical substance is an important scientific question. Rhodococcus ruber SD3, which is a bacterium with organic solvents tolerance, is taken as the subject investigated in the project. Diguanylate cyclase interacting protein and c-di-GMP effector in Rhodococcus ruber SD3 are screened and validated.The role of ligand binding domain of diguanylate cyclase interacting protein is studied. The activity of diguanylate cyclase in nanodisc is assayed and the bioinformatic analysis of c-di-GMP effector is performed. Furthermore,the genes of these two proteins are knocked-out and retro-complemented and the effects of chemical substances on the corresponding mutants are assessed, which will clarify the function of these two proteins. The project deepens the understanding regarding diguanylate cyclase mediated signal pathway in Rhodococcus ruber SD3. At the same time,it has important practical significance in bioprocess of preparation and bioremediation of environmental pollutants.