Gene expression profiling of Pseudomonas putida F1 after exposure to aromatic hydrocarbon in soil by using proteome analysis
Gene expression profiling of Pseudomonas putida F1 after exposure to aromatic hydrocarbon in soil by using proteome analysis
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DOI：
10.1007/s00203-013-0932-4
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发表时间：
2013-10
影响因子：
2.8
通讯作者：
Hajime Morimoto;Masayoshi Kuwano;Yasuhiro Kasahara
中科院分区：
文献类型：
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作者：
Hajime Morimoto;Masayoshi Kuwano;Yasuhiro Kasahara
恶臭假单胞菌F1能代谢甲苯、乙苯和苯生长。在此之前，我们发现了参与这些化合物利用的蛋白质BYP。通过液体培养产生恶臭F1。然而，目前尚不清楚实验室对细菌活性和分解代谢的分析是否准确地反映了土壤环境。我们利用双向凝胶电泳法(2-DE)或标准的SDS-PAGE与液相色谱-串联质谱仪(LC-MS/MS)相结合的方法鉴定了与甲苯、乙苯的降解和土壤中苯的生长有关的蛋白质。根据2-DE/LC-MS/MS分析，在甲苯、乙苯和苯的降解过程中检测到22个关键酶中的12个。在含有乙苯的土壤的标准SDS-PAGE/LC-MS/MS分析中，鉴定出约1,260种细胞蛋白。恶臭F1。所有涉及乙苯降解的关键酶、转运蛋白和感受器蛋白的表达上调，与液体培养中发现的类似。INP。恶臭F1，土壤中的芳香烃响应与在液体介质中观察到的相同。
Pseudomonas putidaF1 can metabolize toluene, ethylbenzene, and benzene for growth. Previously, we identified proteins involved in the utilization of these compounds byP. putidaF1 through culture in liquid media. However, it was unclear whether laboratory analysis of bacterial activity and catabolism accurately reflected the soil environment. We identified proteins involved in the degradation of toluene, ethylbenzene, and benzene growth in soil using two-dimensional gel electrophoresis (2-DE) or standard SDS-PAGE combined with liquid chromatography–tandem mass spectrometry (LC–MS/MS). According to 2-DE/LC–MS/MS analysis, 12 of 22 key enzymes involved in the degradation of toluene, ethylbenzene, and benzene were detected. In standard SDS-PAGE/LC–MS/MS analysis of soil with ethylbenzene, approximately 1,260 cellular proteins were identified inP. putidaF1. All key enzymes and transporter and sensor proteins involved in ethylbenzene degradation were up-regulated similar to that noted in liquid cultures. InP. putidaF1, aromatic hydrocarbon response in soil is the same as that observed in liquid media.