Properties of the hydrogenase of Megasphaera elsdenii.
The catalytic activities of Megasphaera elsdenii hydrogenase are stimulated by salts. The stimulation is due to the anion: the more chaotropic the anion, the greater the effect. Dithionite-reduced and dye-oxidised preparations of hydrogenase are inactivated by reaction with oxygen. The inactivation of the reduced enzyme by excess oxygen follows pseudo-first-order kinetics; the reaction order for the oxidised enzyme has not been established. The rate of oxygen-inactivation is decreased by bovine serum albumin. The hydrogen production activity decreases in the presence of dimethylsulphoxide and ethylene glycol. The hydrogen oxidation activity is stimulated by dimethylsulphoxide, and the activity remains linear with time at concentrations up to 50% (v/v). Above 70% dimethylsulphoxide the steady-state activity of hydrogenase is abolished for both types of activity. The enzyme is more stable in a hydrogen atmosphere than in an argon atmosphere, and the oxidized enzyme is more stable than the reduced enzyme. The enzyme is isolated in the presence of dithionite and it is therefore reduced. When the enzyme is oxidized by treatment with 2,6-dichloroindophenol or with (bi)sulphite, its activity increases by up to 65%; this activation is not reversed when the enzyme is re-reduced. The increase in activity is associated with a change of the redox potential of the incubation medium to a less negative value; half of the maximum activation occurs at -0.41 V. The electron paramagnetic resonance spectrum of the dithionite-reduced hydrogenase resembles that of a reduced ferredoxin-type of spectrum with two 4Fe-4S clusters. The spectrum of the oxidized enzyme is similar to that of Chromatium high-potential iron-sulphur protein. No redox potentials can be ascribed to these spectra since the redox system changes upon freezing to liquid helium temperatures.
D000332:Aerobiosis; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D004355:Drug Stability; D004789:Enzyme Activation; D005026:Ethylene Glycols / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D006864:Hydrogenase; D007700:Kinetics; D009994:Osmolar Concentration; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000378:metabolism*; D012492:Salts / Q000494:pharmacology; D045854:Veillonellaceae / Q000201:enzymology*
D005026:Ethylene Glycols; D012492:Salts; D010088:Oxidoreductases; D006864:Hydrogenase; D004121:Dimethyl Sulfoxide