Probing the proteome response to toluene exposure in the solvent tolerant Pseudomonas putida S12.
Probing the proteome response to toluene exposure in the solvent tolerant Pseudomonas putida S12.
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DOI：
10.1021/pr100401n
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发表时间：
2011-02
影响因子：
4.4
通讯作者：
D. Wijte;B. V. van Baar;A. Heck;A. Altelaar
中科院分区：
文献类型：
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作者：
D. Wijte;B. V. van Baar;A. Heck;A. Altelaar
为了提高生物催化剂的目标产量，有必要彻底了解生产、降解以及重要的适应所需环境的分子机制。其中一种具有很高生物催化潜力的细菌是耐溶剂细菌恶臭假单胞菌S12，它能够降解有机溶剂。为了使有机溶剂的生物转化成为一个成功的工业过程，了解分子对溶剂耐受性的响应是至关重要的。在这里，我们对恶臭假单胞菌S12蛋白质组在甲苯适应的不同阶段进行了定量分析。使用稳定的同位素二甲基化标记方法，我们监测了528个蛋白质的差异表达，包括通常难以检测的膜相关蛋白，如多个RND家族转运蛋白和ABC营养转运蛋白。我们的定量蛋白质组学方法揭示了恶臭假单胞菌S12在甲苯暴露时严重改变其蛋白质表达谱的显著能力。这种蛋白质组的反应导致能量代谢和溶剂外排泵SrpABC表达的显著增加，证实了它在溶剂耐受性中的作用。其他在甲苯存在下强烈上调的蛋白质包括多药外排膜蛋白PP1272和阳离子/醋酸酯共转运体ACTP，它们可能成为改善耐溶剂性的有趣的替代靶点。
To enhance target production from biocatalysts, it is necessary to thoroughly understand the molecular mechanisms involved in production, degradation, and, importantly, adaptation to the required environment. One such bacterium with high potential for biocatalysis is the solvent-tolerant bacteria Pseudomonas putida S12, which, among others, is able to degrade organic solvents. For bioconversion of organic solvents to become a successful industrial process, the understanding of the molecular response upon solvent tolerance is essential. Here we performed a quantitative analysis of the P. putida S12 proteome at different stages of adaptation to toluene. Using a stable isotope dimethylation labeling approach we monitored the differential expression of 528 proteins, including often hard-to-detect membrane associate proteins, such as multiple RND-family transporters and ABC transporters of nutrients. Our quantitative proteomics approach revealed the remarkable ability of P. putida S12 to severely change its protein expression profile upon toluene exposure. This proteome response entails a significant increase in energy metabolism and expression of the solvent efflux pump SrpABC, confirming its role in solvent tolerance. Other proteins strongly up-regulated in the presence of toluene include the multidrug efflux membrane protein PP1272 and the cation/acetate symporter ActP and may form interesting alternative targets for improving solvent tolerance.