Evaluation of rate constants for enzyme-catalysed reactions by the jackknife technique. Application to liver alcohol dehydrogenase

Abstract

Steady-state measurements of enzyme-catalyzed reactions provide more information about the rate constants of the individual steps than is commonly obtained. A combination of the jackknife and non-linear regression techniques were applied to measurements of the rate of oxidation of ethanol by NAD+, catalyzed by alcohol dehydrogenase [EC 1.1.1.1] from horse liver. This has permitted values and confidence intervals to be assigned to the 8 rate constants that characterize the binding of ethanol and NAD+ in random order to the enzyme, and to the net rate constant kcat for the breakdown of the ternary complex.