An Enzyme Kinetics Experiment for the Undergraduate Organic Chemistry Laboratory

An experiment using 1H NMR spectroscopy to observe the kinetics of the acylase 1-catalyzed hydrolysis of N-acetyl-DL-methionine has been developed for the organic laboratory. The L-enantiomer of the reactant is hydrolyzed completely in less than 2 h, and 1H NMR spectroscopic data from a single sample can be worked up to generate Michaelis-Menten and Lineweaver-Burke kinetic plots. The experiment introduces organic chemistry students to the use of a 1H NMR spectrometer, serves as an introduction to enzyme kinetics, and illustrates the stereospecificity of enzyme catalysis. © 2010 American Chemical Society and Division of Chemical Education, Inc.