title: Identification and characterization of a succinyl-coenzyme A (CoA):benzoate CoA transferase in Geobacter metallireducens.
abstract: Geobacter metallireducens is a Fe(III)-respiring deltaproteobacterium and serves as a model organism for aromatic compound-degrading, obligately anaerobic bacteria. In this study, a genetic system was established for G. metallireducens using nitrate as an alternative electron acceptor. Surprisingly, disruption of the benzoate-induced bamY gene, encoding a benzoate coenzyme A (CoA) ligase, reproducibly showed an increased biomass yield in comparison to the wild type during growth with benzoate but not during growth with acetate. Complementation of bamY in trans converted the biomass yield back to the wild-type level. Growth of the bamY mutant with benzoate can be rationalized by the identification of a previously unknown succinyl-CoA:benzoate CoA transferase activity; it represents an additional, energetically less demanding mode of benzoate activation. The activity was highly enriched from extracts of cells grown on benzoate, yielding a 50-kDa protein band; mass spectrometric analysis identified the corresponding benzoate-induced gene annotated as a CoA transferase. It was heterologously expressed in Escherichia coli and characterized as a specific succinyl-CoA:benzoate CoA transferase. The newly identified enzyme in conjunction with a benzoate-induced succinyl-CoA synthetase links the tricarboxylic acid cycle to the upper benzoyl-CoA degradation pathway during growth on aromatic growth substrates.
authors.affiliation: Institute of Biochemistry, University of Leipzig, Leipzig, Germany.
authors.forename: Jana
authors.initials: J
authors.lastname: Oberender
authors.forename: Johannes W
authors.initials: JW
authors.lastname: Kung
authors.forename: Jana
authors.initials: J
authors.lastname: Seifert
authors.forename: Martin
authors.initials: M
authors.lastname: von Bergen
authors.forename: Matthias
authors.initials: M
authors.lastname: Boll
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology*; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology*; D045302:Geobacter / Q000201:enzymology* / Q000235:genetics*; D006841:Hydrocarbons, Aromatic / Q000378:metabolism; D009154:Mutation; D009566:Nitrates / Q000378:metabolism; D013379:Substrate Specificity; D013997:Time Factors; D014166:Transferases / Q000235:genetics / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D006841:Hydrocarbons, Aromatic; D009566:Nitrates; D014166:Transferases