Influence of the fusion of two subunits of the F420-non-reducing hydrogenase of Methanococcus voltae on its biochemical properties.
In Methanococcus voltae, one of the two [NiFeSe] hydrogenases is unusual in that the large subunit is split into two subunits, each contributing two ligands to the [NiFe] center that catalyzes the heterolytic cleavage of the dihydrogen molecule. We have engineered a fusion of these two subunits. The resulting new enzyme showed no significant difference in hydrogen uptake activity or in the Ni-C or Ni-L EPR spectra compared to the the wild-type enzyme, but exhibited a tenfold increase in both the Km for hydrogen and the Ki for the competitive inhibitor carbon monoxide.
Bingemann
R
R
Philipps-Universität Marburg, Fachbereich Biologie, Molekulargenetik, Germany.
Pierik
A J
AJ
Klein
A
A
D001665:Binding Sites; D004578:Electron Spin Resonance Spectroscopy / Q000379:methods; D006859:Hydrogen / Q000378:metabolism; D006864:Hydrogenase / Q000235:genetics / Q000378:metabolism*; D007700:Kinetics; D017017:Methanococcus / Q000201:enzymology* / Q000235:genetics; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism*
D006859:Hydrogen; D010088:Oxidoreductases; D006864:Hydrogenase; C033895:coenzyme F420 hydrogenase