A rapid method for determining kinetic parameters of enzymes exhibiting nonlinear thermal inactivation behavior.
A rapid method is developed to analyze the kinetics of thermal inactivation of enzymes that exhibit a nonlinear biphasic log(activity)-time relationship. Thermal destruction experiments on alcohol dehydrogenase from baker's yeast demonstrate the applicability of the method. The method is based on physical considerations (as opposed to mathematical curve fitting/regression methods) and also serves as a quick check of results obtained using nonlinear regression. It is superior to fitting nonlinear enzyme inactivation data by first-order kinetics or taking the initial and final slopes of the inactivation data. In fact, the method is of general validity and can be applied to any decay process that can be represented by a sum of exponentials.
Nath
S
S
Department of Biochemical Engineering and Biotechnology, Indian Institute of Technology, Hauz Khas, New Delhi 110 016, India.