OPTIMIZATION OF THE COUPLED ENZYMATIC MEASUREMENT OF SUBSTRATE

In the kinetic analysis of substrate, factors which affect the enzymatic activity will affect the estimate of the amount of substrate. Likewise variations in the activity of coupling enzymes will affect the estimate. By proper selection of the time at which to measure the rate, variations in enzymatic activities will cause only minor variations in the rate, contrary to the case where measurements of the initial rate are made. Optimization theory is developed for variations in the activity of the primary or the coupling or both enzymes. Four methods are presented for estimating the kinetic parameters used in the optimization. © 1979, American Chemical Society. All rights reserved.