title: Dependence on the F0F1-ATP synthase for the activities of the hydrogen-oxidizing hydrogenases 1 and 2 during glucose and glycerol fermentation at high and low pH in Escherichia coli.
abstract: Escherichia coli has four [NiFe]-hydrogenases (Hyd); three of these, Hyd-1, Hyd-2 and Hyd-3 have been characterized well. In this study the requirement for the F(0)F(1)-ATP synthase for the activities of the hydrogen-oxidizing hydrogenases Hyd-1 and Hyd-2 was examined. During fermentative growth on glucose at pH 7.5 an E. coli F(0)F(1)-ATP synthase mutant (DK8) lacked hydrogenase activity. At pH 5.5 hydrogenase activity was only 20% that of the wild type. Using in-gel activity staining, it could be demonstrated that both Hyd-1 and Hyd-2 were essentially inactive at these pHs, indicating that the residual activity at pH 5.5 was due to the hydrogen-evolving Hyd-3 enzyme. During fermentative growth in the presence of glycerol, hydrogenase activity in the mutant was highest at pH 7.5 attaining a value of 0.76 U/mg, or ~50% of wild type activity, and Hyd-2 was only partially active at this pH, while Hyd-1 was inactive. Essentially no hydrogenase activity was measured at pH 5.5 during growth with glycerol. At this pH the mutant had a hydrogenase activity that was maximally only ~10% of wild type activity with either carbon substrate but a weak activity of both Hyd-1 and Hyd-2 could be detected. Taken together, these results demonstrate for the first time that the activity of the hydrogen-oxidizing hydrogenases in E. coli depends on an active F(0)F(1)-ATP synthase during growth at high and low pH.
authors.affiliation: Department of Biophysics, Yerevan State University, 1 A. Manoukian Str, 0025, Yerevan, Armenia.
authors.forename: Karen
authors.initials: K
authors.lastname: Trchounian
authors.forename: Constanze
authors.initials: C
authors.lastname: Pinske
authors.forename: R Gary
authors.initials: RG
authors.lastname: Sawers
authors.forename: Armen
authors.initials: A
authors.lastname: Trchounian
mesh_terms: D003451:Cryoprotective Agents / Q000378:metabolism / Q000494:pharmacology; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D005285:Fermentation / Q000502:physiology*; D005947:Glucose / Q000378:metabolism* / Q000494:pharmacology; D005990:Glycerol / Q000378:metabolism* / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D009154:Mutation; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism*; D006180:Proton-Translocating ATPases / Q000235:genetics / Q000378:metabolism*; D013549:Sweetening Agents / Q000378:metabolism / Q000494:pharmacology
chemical_list: D003451:Cryoprotective Agents; D029968:Escherichia coli Proteins; D013549:Sweetening Agents; D010088:Oxidoreductases; C029746:uptake hydrogenase; D006180:Proton-Translocating ATPases; D005947:Glucose; D005990:Glycerol
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