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Escherichia coli hydrogenase 3 is a reversible enzyme possessing hydrogen uptake and synthesis activities

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Abstract

In the past, it has been difficult to discriminate between hydrogen synthesis and uptake for the three active hydrogenases in Escherichia coli (hydrogenase 1, 2, and 3); however, by combining isogenic deletion mutations from the Keio collection, we were able to see the role of hydrogenase 3. In a cell that lacks hydrogen uptake via hydrogenase 1 (hyaB) and via hydrogenase 2 (hybC), inactivation of hydrogenase 3 (hycE) decreased hydrogen uptake. Similarly, inactivation of the formate hydrogen lyase complex, which produces hydrogen from formate (fhlA) in the hyaB hybC background, also decreased hydrogen uptake; hence, hydrogenase 3 has significant hydrogen uptake activity. Moreover, hydrogen uptake could be restored in the hyaB hybC hycE and hyaB hybC fhlA mutants by expressing hycE and fhlA, respectively, from a plasmid. The hydrogen uptake results were corroborated using two independent methods (both filter plate assays and a gas-chromatography-based hydrogen uptake assay). A 30-fold increase in the forward reaction, hydrogen formation by hydrogenase 3, was also detected for the strain containing active hydrogenase 3 activity but no hydrogenase 1 or 2 activity relative to the strain lacking all three hydrogenases. These results indicate clearly that hydrogenase 3 is a reversible hydrogenase.

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Acknowledgment

The authors would like to thank the National Institute of Genetics, Japan for sending the Keio and ASKA clones. This research was supported by DARPA (HR0011-06-1-0001).

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Authors and Affiliations

  1. Artie McFerrin Department of Chemical Engineering, Texas A & M University, 220 Jack E. Brown Building, College Station, TX, 77843-3122, USA

    Toshinari Maeda, Viviana Sanchez-Torres & Thomas K. Wood

  2. Department of Biology, Texas A & M University, College Station, TX, 77843-3258, USA

    Thomas K. Wood

  3. Zachry Department of Civil and Environmental Engineering, Texas A & M University, College Station, TX, 77843-3136, USA

    Thomas K. Wood

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  1. Toshinari Maeda
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  2. Viviana Sanchez-Torres
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  3. Thomas K. Wood
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Correspondence to Thomas K. Wood.

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Maeda, T., Sanchez-Torres, V. & Wood, T.K. Escherichia coli hydrogenase 3 is a reversible enzyme possessing hydrogen uptake and synthesis activities. Appl Microbiol Biotechnol 76, 1035–1042 (2007). https://doi.org/10.1007/s00253-007-1086-6

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