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Keywords:

  • biocatalysis;
  • EPR spectroscopy;
  • FTIR spectroscopy;
  • hydrogenases;
  • oxygen tolerance
Thumbnail image of graphical abstract

A clear picture: In situ EPR and FTIR spectroscopic studies on the soluble, NAD+-reducing [NiFe]-hydrogenase of Ralstonia eutropha reveal that the catalytic site resides predominantly in the intermediate Nia-C state within whole cells. This state, can either be reversibly oxidized to a “Nir-B”-like state or further reduced to various Nia-SR species. The data suggest that the iron center in the active site contains a standard set (one CO and two CN) of inorganic ligands.