Estimating the Product Inhibition Constant from Enzyme Kinetic Equations Using the Direct Linear Plot Method in One-Stage Treatment

Abstract

A direct linear plot was applied to estimate kinetic constants using the product’s competitive inhibition equation. The challenge consisted of estimating three kinetic constants, Vmax, Km, and Kp, using two independent variables, substrates, and product concentrations, in just one stage of mathematical treatment. The method consisted of combining three initial reaction rate data and avoiding the use of the same three product concentrations (otherwise, this would result in a mathematical indetermination). The direct linear plot method was highly superior to the least-squares method in terms of accuracy and robustness, even under the addition of error. The direct linear plot method is a reliable and robust method that can be applied to estimate Kp in inhibition studies in pharmaceutical and biotechnological areas.

Keywords

• direct linear plot
• median method
• product inhibition
• kinetic constants
• non-parametric
• distribution-free method