Enzyme Kinetics Experiment with the Multienzyme Complex Viscozyme L and Two Substrates for the Accurate Determination of Michaelian Parameters

摘要

A laboratory experiment in which students study the kinetics of the Viscozyme-L-catalyzed hydrolysis of cellulose and starch comparatively was designed for an upperdivision biochemistry laboratory. The main objective of this experiment was to provide an opportunity to perform enhanced enzyme kinetics data analysis using appropriate informatics and statistical tools. By fitting the Michaelis Menten, Lineweaver-Burk, Eadie-Hofstee, and Hanes equations to the experimental data, students explore the strengths and weaknesses of each plot and determine whether the two Viscozyme-L-catalyzed reactions obey Michaelis Menten kinetics. Comparing the values of the Michaelian constants (nu(max) and K-M) and the efficiencies obtained for each enzyme-catalyzed reaction allows advanced undergraduates to determine if the differences between. these parameters are statistically significant.