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

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

Research output: Contribution to journalArticle

70 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)1035-1042
Number of pages8
JournalApplied Microbiology and Biotechnology
Volume76
Issue number5
DOIs
StatePublished - Oct 1 2007

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Hydrogenase
Escherichia coli
Hydrogen
Enzymes
formic acid
Assays
Sequence Deletion
Gas chromatography
Gas Chromatography
Plasmids

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Applied Microbiology and Biotechnology

Cite this

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Escherichia coli hydrogenase 3 is a reversible enzyme possessing hydrogen uptake and synthesis activities. / Maeda, Toshinari; Sanchez-Torres, Viviana; Wood, Thomas K.

In: Applied Microbiology and Biotechnology, Vol. 76, No. 5, 01.10.2007, p. 1035-1042.

Research output: Contribution to journalArticle

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