An optimum regimen for the use of malate dehydrogenase and lactate dehydrogenase during chemical regeneration of the cofactor
Markina, V.L.; Eremin, A.N.; Metelitsa, D.I.
Prikladnaia Biokhimiia i Mikrobiologiia 22(5): 635-641
1986
ISSN/ISBN: 0555-1099 PMID: 3534877 Document Number: 279648
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.