Comparative analysis on the membrane proteome of Clostridium acetobutylicum wild type strain and its butanol-tolerant mutant
Comparative analysis on the membrane proteome of Clostridium acetobutylicum wild type strain and its butanol-tolerant mutant
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DOI：
10.1039/c0mb00330a
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发表时间：
2011-01-01
影响因子：
--
通讯作者：
Ma, Yanhe
中科院分区：
文献类型：
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作者：
Mao, Shaoming;Luo, Yuanming;Ma, Yanhe
产溶剂细菌丙酮丁醇梭菌是发酵工业中使用的最重要的梭菌属物种。然而，对丁醇的不耐受性阻碍了溶剂的有效生产。丁醇的毒性主要是由于丁醇对细胞膜的离液作用，但对丁醇对膜相关蛋白的影响的认识非常有限。采用2-DE结合MALDI-TOF MS/MS和1-DE结合LC-MS/MS的方法，对细胞裂解液膜组分中的341个蛋白质进行了鉴定，建立了第一个完整的C.丙酮丁醇已鉴定的蛋白质主要参与运输、细胞膜/壁机械、表面被膜和鞭毛的形成以及能量代谢。对野生型菌株DSM 1731及其丁醇耐受突变株Rh 8的膜蛋白质组进行比较分析，发现73个差异表达蛋白。层次聚类分析表明，突变体Rh 8可能已经进化出一个更稳定的膜结构，并已开发出一个成本效益的能量代谢策略，以科普丁醇的挑战。这项比较膜蛋白质组学研究，加上我们以前发表的比较细胞质蛋白质组学的工作，使我们能够获得丁醇对C.丙酮丁醇
The solventogenic bacterium Clostridium acetobutylicum is the most important species of Clostridium used in the fermentation industry. However, the intolerance to butanol hampers the efficient production of solvents. Butanol toxicity has been attributed to the chaotropic effect on the cell membrane, but the knowledge on the effect of butanol on membrane associated proteins is quite limited. Using 2-DE combined with MALDI-TOF MS/MS and 1-DE integrated with LC-MS/MS, 341 proteins in the membrane fractions of cell lysate were identified, thus establishing the first comprehensive membrane proteome of C. acetobutylicum. The identified proteins are mainly involved in transport, cellular membrane/wall machinery, formation of surface coat and flagella, and energy metabolism. Comparative analysis on the membrane proteomes of the wild type strain DSM 1731 and its butanol-tolerant mutant Rh8 revealed 73 differentially expressed proteins. Hierarchical clustering analysis suggested that mutant Rh8 may have evolved a more stabilized membrane structure, and have developed a cost-efficient energy metabolism strategy, to cope with the butanol challenge. This comparative membrane proteomics study, together with our previous published work on comparative cytoplasmic proteomics, allows us to obtain a systemic understanding of the effect of butanol on cellular physiology of C. acetobutylicum.