TABLE 1.

Technical and conceptual learning goals of the E and S protocols

Technical learning goals specific to the E protocol

Learn to use volumetric equipment, such as reagent dispenser bottles, graduated cylinders, and pipettes to mix reagent stocks of buffer, substrate, enzyme, and inhibitors Learn to use strong alkali (NaOH) to stop the enzymatic reaction and increase the specific absorbance of the reaction product, p-nitrophenol. Learn to control the pH of solution, and accurately time the duration of the assay (e.g., 15 minutes). Learn to use a temperature-controlled water bath. Learn the Beer-Lambert law of light absorbance and how to use a spectrophotometer to measure absorbance and the amount of a chemical (p-nitrophenol) dissolved in water. Learn various laboratory psychomotor skills important for a profession in the life sciences.

Technical learning goals specific to the S protocol

Learn to use a graphic user interface (GUI) to control reaction parameters: pH, temperature, reaction time, and chemical concentrations (enzyme, substrate, inhibitors) Learn to use a GUI to select preset simulation parameters: elementary rate constants for forward and reverse binding reactions for substrate and inhibitors; enzyme turnover rate. Learn to read out the amount of product made predicted by the simulation and interpret concentration versus time plots for all chemical species included in the Michaelis-Menten model.

Conceptual learning goals common to both protocols

Learn basic concepts of experimental protocol design and execution. Learn to calculate the activity of an enzyme from the amount of product made during a fixed period (assay time). Learn to find the optimum pH and temperature of an enzyme-catalyzed reaction. Learn to calculate the enzyme’s kinetic parameters Vmax and KM for a given substrate and how reversible chemical inhibitors can be used to manipulate those parameters.