In this laboratory, you will perform simulations of experiments designed to
study the biochemistry of the enzyme invertase, an important enzyme involved
in the metabolism of the disaccharide sucrose. You will learn to measure and
calculate important parameters of enzyme kinetics, and to measure product formation
by spectrophotometry. Also, by changing reaction conditions and plotting enzyme
kinetic data, you will learn about factors that influence the catalytic activity
The purpose of this laboratory is to:
- Help you understand important properties of enzyme-catalyzed
reactions, and techniques that are used to measure enzymatic reactions.
- Demonstrate how changes in important parameters of
enzyme biochemistry, such as substrate concentration, can affect enzyme
- Study and measure MichaelisMenton enzyme kinetics.
- Investigate pH and temperature optimums of invertase.
- Examine the effects of different classes of enzyme
inhibitors on invertase.
- Simulate the spectrophotometric measurement of enzyme-catalyzed
Before You Begin: Prerequisites
Before beginning EnzymeLab you should be familiar with the
- Basic principles of thermodynamics, metabolism, and metabolic
- The structure of sucrose and its importance as a source
- Properties and functions of protein enzymes as biological
- Temperature and pH as important factors that affect enzyme
- Feedback mechanisms involved in the control of enzyme
activity and metabolism, and different types of enzyme inhibitors including
competitive inhibitors and noncompetitive inhibitors.
- First-order enzyme kinetics measurements of the MichaelisMenten
equation including VO, Vmax, [S], and KM.
- Analysis of kinetic data by the following plots: V vs.
[S], V vs. pH, V vs. temperature, LineweaverBurk, and EadieHofstee
- Basic operations of a visible light spectrophotometer
and its application in measuring enzyme kinetics.