ABSTRACT
The majority of this book has dealt with situations where the enzyme concentration is less than that of the substrate, so that a steady state exists during the reaction. Steady-state kinetic studies can be used to determine the order of addition and release of reactants and products and where the major rate-limiting steps are in a mechanism. However, one gets only limited information on isomerization of transitory intermediates after the substrates add. Isotope effects permit the determination of partition ratios for intermediates, and they and pH profiles give evidence of the chemistry that is taking place. To determine the intrinsic rate constants and to characterize intermediates usually requires pre-steady-state experiments.
