[An optimum regimen for the use of malate dehydrogenase and lactate dehydrogenase during chemical regeneration of the cofactor].
The steady-state kinetics of malate oxidation by malate dehydrogenase was being studied without coupling reagents under the conditions of chemical regeneration of the cofactor by the following pairs: phenazine methosulphate (PMS)--dichlorphenolindophenol (DCPIP) and PMS--tetranitrotetrazolium blue (TNTB). The comparative kinetic study was carried out of the steady-state oxidation of lactate and the reduction of pyruvate by lactate rehydrogenase, as well as of the dehydrogenation of lactate, coupled with the cofactor regeneration by the pair PMS-DCPIP. Optimum reagent concentrations, optimum pH and activation energies were determined for six systems. Malate dehydrogenation coupled with regeneration of the cofactor by the pair PMS-TNTB is the most promising reaction for enzyme immunoassay.
D000818:Animals; D007124:Immunoenzyme Techniques; D007700:Kinetics; D007770:L-Lactate Dehydrogenase / Q000378:metabolism*; D008291:Malate Dehydrogenase / Q000378:metabolism*; D008929:Mitochondria, Heart / Q000201:enzymology; D009243:NAD / Q000378:metabolism*; D010084:Oxidation-Reduction; D013552:Swine
D009243:NAD; D007770:L-Lactate Dehydrogenase; D008291:Malate Dehydrogenase
Optimal'nyĭ rezhim ispol'zovaniia malatdegidrogenazy i laktatdegidrogenazy v usloviiakh khimicheskoĭ regeneratsii kofaktora.