Probing the proteome response to toluene exposure in the solvent tolerant Pseudomonas putida S12.
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.
TNO Defence, Security and Safety, Business Unit Biological and Chemical Protection, Rijswijk, The Netherlands.
Dorien
D
Wijte
Ben L M
BL
van Baar
Albert J R
AJ
Heck
A F Maarten
AF
Altelaar
D001426:Bacterial Proteins / Q000032:analysis / Q000378:metabolism*; D055162:Biocatalysis; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D016000:Cluster Analysis; D004591:Electrophoresis, Polyacrylamide Gel; D007218:Industrial Microbiology; D007553:Isotope Labeling; D053858:Metabolic Networks and Pathways / Q000187:drug effects; D010446:Peptide Fragments / Q000032:analysis / Q000378:metabolism; D020543:Proteome / Q000378:metabolism*; D016958:Pseudomonas putida / Q000187:drug effects* / Q000378:metabolism; D014050:Toluene / Q000494:pharmacology*; D014357:Trypsin
D001426:Bacterial Proteins; D010446:Peptide Fragments; D020543:Proteome; D014050:Toluene; D014357:Trypsin