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通过定量蛋白质组学揭示苯酚降解菌株假单胞菌属M1对苯酚全局响应的机制性见解。

Mechanistic insights into the global response to phenol in the phenol-biodegrading strain Pseudomonas sp. M1 revealed by quantitative proteomics.

作者信息

Santos Pedro M, Roma Vasco, Benndorf Dirk, von Bergen Martin, Harms Hauke, Sá-Correia Isabel

机构信息

Institute for Biotechnology and Bioengineering, Centre for Biological and Chemical Engineering, Instituto Superior Técnico, Lisboa, Portugal.

出版信息

OMICS. 2007 Fall;11(3):233-51. doi: 10.1089/omi.2007.0009.

Abstract

Quantitative proteomics was used to gain insights into the global adaptive response to phenol in the phenol-biodegrading strain Pseudomonas sp. M1 when an alternative carbon source (pyruvate or succinate) is present. A phylogenetic analysis indicated Pseudomonas citronellolis as the closest species to the environmental strain M1, while P. aeruginosa is the closest species with the genome sequence available. After two-dimensional gel electrophoresis (2-DE) separation, protein identification by MS/MS ion search allowed the assignment of 87 out of 136 selected protein spots, 56 of which matched P. aeruginosa proteins present in databases. Coordinate induction of six enzymes of the phenol catabolic pathway in cells grown in pyruvate and phenol was revealed by expression proteomics. When succinate was the alternative carbon source (C-source), these catabolic proteins were not expressed. The global response of Pseudomonas sp. M1 to phenol-induced stress involved, among others, proteins of the energy metabolism, stress response proteins, and transport proteins. Quantitative and/or qualitative differences were registered in M1 response to different phenol concentrations or to identical phenol concentrations when cells were grown in pyruvate or succinate medium. They were attributed to differences observed in the specific growth rate, in the expression of phenol catabolism, and in resistance to phenol of Pseudomonas sp. M1 grown under different conditions.

摘要

当存在替代碳源(丙酮酸或琥珀酸)时,采用定量蛋白质组学方法深入了解苯酚生物降解菌株假单胞菌属M1对苯酚的全局适应性反应。系统发育分析表明,香茅假单胞菌是与环境菌株M1亲缘关系最近的物种,而铜绿假单胞菌是具有可用基因组序列的亲缘关系最近的物种。经过二维凝胶电泳(2-DE)分离后,通过MS/MS离子搜索进行蛋白质鉴定,可对136个选定蛋白质斑点中的87个进行归属,其中56个与数据库中存在的铜绿假单胞菌蛋白质相匹配。表达蛋白质组学揭示了在丙酮酸和苯酚中生长的细胞中苯酚分解代谢途径的六种酶的协同诱导。当琥珀酸作为替代碳源(C源)时,这些分解代谢蛋白不表达。假单胞菌属M1对苯酚诱导的应激的全局反应尤其涉及能量代谢蛋白、应激反应蛋白和转运蛋白。当细胞在丙酮酸或琥珀酸培养基中生长时,M1对不同苯酚浓度或相同苯酚浓度的反应存在定量和/或定性差异。这些差异归因于在不同条件下生长的假单胞菌属M1在比生长速率、苯酚分解代谢表达和对苯酚的抗性方面观察到的差异。

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