Monday, July 30, 2007 - 1:30 PM
S53

Proteomic investigation of alcohol and ketone metabolism in Sulfolobus solfataricus P2

Poh Koh Chong1, Adam M. Burja2, Helia Radianingtyas2, and Phillip C. Wright3. (1) Chemical and Process Engineering, University of Sheffield, Mappin Street, Sheffield, United Kingdom, (2) Metabolic Engineering and Fermentation, Ocean Nutrition Canada, 101 Research Drive, Dartmouth, NS B2Y 4T6, Canada, (3) Department of Chemical and Process Engineering, Biological and Environmental Systems Group, The University of Sheffield, Mappin Street, Sheffield, England, S1 3JD, United Kingdom

The proteomic response of Sulfolobus solfataricus P2, a hyperthermophilic archeon, to different concentrations of 1- and 2-propanol, ethanol, phenol and acetone as sole carbon sourcess was investigated. Results were compared to growth on glucose alone, and with glucose as a co-substrate. A particular focus was directed on ethanol as the sole carbon source, with feed concentrations of 0.79 to 39.8g/L. Ethanol uptake was confirmed using GC/MS, and in vivo metabolic labelling using 13C universally labelled ethanol and LC-MS/MS. To obtain deeper mechanistic understanding, a proteomic approach was employed using both 2-dimensional electrophoresis (2-DE) and quantitiative shotgun proteomics employing isobaric mass tags (iTRAQ). From these workflows, ca. 22% and 35% of the identified and quantified 352 proteins were up- or down-regulated respectively, compared to the glucose control. Examples include a greater than 2 fold up-regulation of the zinc dependent alcohol dehydrogenase, adh-10 (SSO2536) and the putative adh-2 (SSO0764). A comparison was made with the corresponding mRNA relative abundance changes (using quantitative RT-PCR), yielding similar fold-changes, thus suggesting an integral role for adh-2 and adh-10 in ethanol metabolism. Overall global data suggested that ethanol was catabolised into central metabolism via acetyl-CoA intermediates.

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