赤红球菌对有机溶剂耐受性的机理研究

耐有机溶剂微生物对于生产一些大宗化学品、全细胞生物催化和一些环境污染物的生物修复都起着重要的作用。目前关于有机溶剂耐受菌的研究大多数集中在革兰氏阴性菌（尤其是假单胞菌和大肠杆菌）。赤红球菌SD3是具有优良有机溶剂耐受性的革兰氏阳性菌。本项目以赤红球菌SD3为材料，利用定量iTRAQ LC-MS/MS 的蛋白质组学技术，分析其在不同有机溶剂（甲苯和苯酚）胁迫下的蛋白质组。在此基础上，选择与赤红球菌SD3有机溶剂耐受性密切相关的一些新颖和重要的蛋白质作为研究对象，在赤红球菌SD3中对相应的基因进行敲除、回补和强化，进一步确定这些蛋白质对赤红球菌有机溶剂耐受性所起的作用。本项目在蛋白质组学水平上系统地探究赤红球菌对有机溶剂的耐受性机理，不仅可以进一步丰富人们对革兰氏阳性菌有机溶剂耐受性的认识，而且还为革兰氏阳性菌的有机溶剂耐受性进行合理改造提供科学方法。

Organic solvents tolerant microorganisms play an important role in producing some bulk chemicals, whole cell biocatalysis and bioremediation of some pollutants. Until now the studies on organic solvents tolerant microorganisms have focused on Gram-negative microbes, especially Pesudomonas and E.coli. Rhodococcus ruber SD3 is a kind of Gram-positive bacterium with good organic solvents tolerance. Proteomes from Rhodococcus ruber SD3 under different orgainic slovents (toluene and phenol) stress are analyzed with iRAQI LC-MS/MS proteomic method in the project. Based on these results, some novel and important proteins related to organic slovents tolerance are selected and the corresponding genes are knock-outed, retro-complemented and reinforced in Rhodococcus ruber SD3 to validate the role of these proteins in organic solvents tolerance. The project systematically studies the organic solvents tolerant mechanisms of Rodococcus ruber in proteomic levels, which not only helps us to further understand organic slovent tolerance of Gram-positive microorganism, but supplies a scientific method for engineering of orgnic solvents tolerance in Gram-positive microorganism.