title: Degradation of acetaldehyde and its precursors by Pelobacter carbinolicus and P. acetylenicus.
abstract: Pelobacter carbinolicus and P. acetylenicus oxidize ethanol in syntrophic cooperation with methanogens. Cocultures with Methanospirillum hungatei served as model systems for the elucidation of syntrophic ethanol oxidation previously done with the lost "Methanobacillus omelianskii" coculture. During growth on ethanol, both Pelobacter species exhibited NAD+-dependent alcohol dehydrogenase activity. Two different acetaldehyde-oxidizing activities were found: a benzyl viologen-reducing enzyme forming acetate, and a NAD+-reducing enzyme forming acetyl-CoA. Both species synthesized ATP from acetyl-CoA via acetyl phosphate. Comparative 2D-PAGE of ethanol-grown P. carbinolicus revealed enhanced expression of tungsten-dependent acetaldehyde: ferredoxin oxidoreductases and formate dehydrogenase. Tungsten limitation resulted in slower growth and the expression of a molybdenum-dependent isoenzyme. Putative comproportionating hydrogenases and formate dehydrogenase were expressed constitutively and are probably involved in interspecies electron transfer. In ethanol-grown cocultures, the maximum hydrogen partial pressure was about 1,000 Pa (1 mM) while 2 mM formate was produced. The redox potentials of hydrogen and formate released during ethanol oxidation were calculated to be EH2 = -358±12 mV and EHCOOH = -366±19 mV, respectively. Hydrogen and formate formation and degradation further proved that both carriers contributed to interspecies electron transfer. The maximum Gibbs free energy that the Pelobacter species could exploit during growth on ethanol was -35 to -28 kJ per mol ethanol. Both species could be cultivated axenically on acetaldehyde, yielding energy from its disproportionation to ethanol and acetate. Syntrophic cocultures grown on acetoin revealed a two-phase degradation: first acetoin degradation to acetate and ethanol without involvement of the methanogenic partner, and subsequent syntrophic ethanol oxidation. Protein expression and activity patterns of both Pelobacter spp. grown with the named substrates were highly similar suggesting that both share the same steps in ethanol and acetalydehyde metabolism. The early assumption that acetaldehyde is a central intermediate in Pelobacter metabolism was now proven biochemically.
authors.affiliation: Department of Biology, University of Konstanz, D-78457, Konstanz, Germany.
authors.forename: Alexander
authors.initials: A
authors.lastname: Schmidt
authors.affiliation: Department of Biology, University of Konstanz, D-78457, Konstanz, Germany.
authors.forename: Marco
authors.initials: M
authors.lastname: Frensch
authors.affiliation: Department of Biology, University of Konstanz, D-78457, Konstanz, Germany.
authors.forename: David
authors.initials: D
authors.lastname: Schleheck
authors.affiliation: Department of Biology, University of Konstanz, D-78457, Konstanz, Germany.
authors.forename: Bernhard
authors.initials: B
authors.lastname: Schink
authors.affiliation: Department of Biology, University of Konstanz, D-78457, Konstanz, Germany.
authors.forename: Nicolai
authors.initials: N
authors.lastname: Müller
mesh_terms: D000079:Acetaldehyde / Q000378:metabolism*; D000105:Acetyl Coenzyme A / Q000378:metabolism; D000426:Alcohol Dehydrogenase / Q000378:metabolism; D001426:Bacterial Proteins / Q000378:metabolism; D018920:Coculture Techniques; D020564:Deltaproteobacteria / Q000201:enzymology / Q000254:growth & development / Q000378:metabolism*; D005560:Formate Dehydrogenases / Q000378:metabolism; D005561:Formates / Q000378:metabolism; D006859:Hydrogen / Q000378:metabolism; D019639:Methanospirillum / Q000201:enzymology / Q000254:growth & development / Q000378:metabolism; D010084:Oxidation-Reduction
chemical_list: D001426:Bacterial Proteins; D005561:Formates; C030544:formic acid; D000105:Acetyl Coenzyme A; D006859:Hydrogen; D000426:Alcohol Dehydrogenase; D005560:Formate Dehydrogenases; D000079:Acetaldehyde