title: Measuring the pH dependence of hydrogenase activities.
abstract: The pH dependences of activities of homogenous hydrogenases of Thiocapsa roseopersicina and Desulfomicrobium baculatum in the reaction of hydrogen uptake in solution in the presence of benzyl viologen and the pH dependences of catalytic currents of hydrogen oxidation by electrodes on which these hydrogenases were immobilized were compared. Maximal activities of the hydrogenases from T. roseopersicina and D. baculatum in the reaction hydrogen uptake in solution were observed at pH 9.5 and 8.5, respectively. However, the steady-state current caused by catalytic uptake of hydrogen was maximal for the T. roseopersicina hydrogenase-containing electrode at pH 5.5-6.5 under overvoltage of 30-60 mV, whereas for electrodes with D. baculatum hydrogenase it was maximal at pH 6.0-6.5. Analysis of these data suggests that pH-dependent changes in the hydrogenase activities in solution during hydrogen uptake are due not only to the effect of proton concentration on the enzyme conformation or protonation of certain groups of the enzyme active center, but they are rather indicative of changes in free energy of the reaction accompanying changes in pH.
authors.affiliation: Institute of Basic Biological Problems, Russian Academy of Sciences, Pushchino, Moscow Region 142290, Russia. ttt@issp.serpukhov.su
authors.forename: A A
authors.initials: AA
authors.lastname: Tsygankov
authors.forename: E A
authors.initials: EA
authors.lastname: Minakov
authors.forename: N A
authors.initials: NA
authors.lastname: Zorin
authors.forename: K S
authors.initials: KS
authors.lastname: Gosteva
authors.forename: O G
authors.initials: OG
authors.lastname: Voronin
authors.forename: A A
authors.initials: AA
authors.lastname: Karyakin
mesh_terms: D001594:Benzyl Viologen / Q000737:chemistry; D001665:Binding Sites; D003901:Desulfovibrio / Q000201:enzymology*; D006859:Hydrogen / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D006864:Hydrogenase / Q000378:metabolism*; D020616:Thiocapsa roseopersicina / Q000201:enzymology*
chemical_list: D001594:Benzyl Viologen; D006859:Hydrogen; D006864:Hydrogenase