13C and 61Ni isotope substitutions confirm the presence of a nickel (III)-carbon species in acetogenic CO dehydrogenases.
The nickel-containing CO dehydrogenases from Acetobacterium woodii and Clostridium thermoaceticum were studied by EPR spectroscopy in order to define the components involved in the EPR spectrum obtained by reaction of the enzymes with the substrate, CO. Using isotopic substitution techniques, these experiments unequivocally establish that a nickel-carbon species is involved in the g = 2.08, 2.02 EPR signal. Comparing the 61Ni- and 59Ni-substituted enzymes, the g = 2.08 component of the resonance was found to be mainly due to nickel with a smaller contribution by the carbon species. Reaction of the CO dehydrogenase with [13C]CO versus [12C]CO showed that a carbon species, formed from CO, was the major contributor to the g = 2.02 EPR signal. In addition, the oxidized CO dehydrogenase was found to exhibit a Ni (III) EPR signal analogous to that of the hydrogenases from the methanogenic and sulfate-reducing bacteria.
D000445:Aldehyde Oxidoreductases; D002244:Carbon; D002247:Carbon Isotopes; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D003013:Clostridium / Q000201:enzymology; D004578:Electron Spin Resonance Spectroscopy; D006094:Gram-Positive Bacteria / Q000201:enzymology; D007554:Isotopes; D009097:Multienzyme Complexes; D009532:Nickel
D002247:Carbon Isotopes; D007554:Isotopes; D009097:Multienzyme Complexes; D002244:Carbon; D009532:Nickel; D000445:Aldehyde Oxidoreductases; C019823:carbon monoxide dehydrogenase