Samueli School of Engineering

The objective of this experiment was to extract a whole cell protein from bacterial cells and perform affinity chromatography in order to characterize the reaction kinetics of a protein enzyme, β-galactosidase. This was performed by varying the volume of substrate and enzyme for each trial in order to study the effects on the reaction rate. The reaction rate is then analyzed using the Michaelis-Menten equation to determine the velocity of the reaction using enzyme concentration. Once the velocity is known, the equation can be linearized to determine the max velocity for the reaction for a given substrate concentration. It is expected that increasing the enzyme concentration or substrate concentration will give a higher reaction rate. As the substrate concentration decrease, the reaction rate will decrease as well. This is important in food industry in fermentation process to reduce lactose levels in dairy products for intolerant people.