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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

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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 F0F1-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 F0F1-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 F0F1-ATP synthase during growth at high and low pH.

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Authors and Affiliations

  1. Department of Biophysics, Yerevan State University, 1 A. Manoukian Str, 0025, Yerevan, Armenia

    Karen Trchounian & Armen Trchounian

  2. Institute for Microbiology, Martin-Luther University Halle-Wittenberg, 06120, Halle, Germany

    Karen Trchounian, Constanze Pinske & R. Gary Sawers

Authors
  1. Karen Trchounian
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  2. Constanze Pinske
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  3. R. Gary Sawers
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  4. Armen Trchounian
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Correspondence to Armen Trchounian.

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Trchounian, K., Pinske, C., Sawers, R.G. et al. 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 . J Bioenerg Biomembr 43, 645–650 (2011). https://doi.org/10.1007/s10863-011-9397-9

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  • DOI: https://doi.org/10.1007/s10863-011-9397-9

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