In the Enzyme Kinetics Lab, you will learn how substrates are converted into products by catalysis. You will also learn all about the kinetics of enzyme involving the Michaelis-Menten equation and various rate constants, as well as DNA mutation and hyperactivity. You will get to run experiments using the enzyme Alcohol Dehydrogenase on a wild and mutant type to learn about Alcohol Flush Syndrome.

Use a spectrophotometer to measure enzyme reaction

In the Enzyme Kinetics lab, you will access a fully equipped workbench where you can prepare the Alcohol Dehydrogenase reaction and measure the product of Acetylaldehyde using a spectrophotometer. You will learn about the concept of spectrophotometry, how to prepare a master mix and how to calculate dilution. You will try to prepare a reaction in a 1 ml cuvette and measure the amount of product formed using the spectrophotometer.

See it all on a molecular level

Supplementary 3D animations illustrate what happens at the molecular level when the substrate and co-factor enter the active site. During the 3D animations, you will also answer quiz questions to test your understanding of the concepts. The animations are interactive, so you can identify the substrate by clicking on the different molecules.

Experiment freely and measure the results

For every measurement, you receive a progress curve displaying amounts of product formed over time. You must then analyze the outcome data and plot your own Michaelis-Menten graph to find the Km and Vmax for each enzyme. By comparing Km and Vmax values of the wild type vs. mutant Alcohol Dehydrogenase, you will be able to understand the Alcohol Flush Syndrome. With the newly added module of enzyme inhibition, you are asked to perform different enzyme inhibition experiments using three different inhibitors. You can measure product formation using several inhibitor concentrations, extract the data, create your own Lineweaver-Burk plot and solve the Ki.

Updated with a mathematically based simulator

We have recently upgraded the Enzyme Kinetics lab by implementing a mathematically based simulator. This provides you with a larger flexibility in conducting the experiments, allowing you to change parameters such as substrate concentrations, enzyme concentrations, temperature or pH and receive the corresponding results. In this semi-guided module, you can experiment with different parameters in order to find the optimal temperature and pH to reach the highest initial reaction rate.

Upon completing the Enzyme Kinetics lab, you will be familiar with the kinetics of enzyme Alcohol Dehydrogenase. Will you be able to use your newly acquired knowledge to perform the experiment and analyze the data outcome? And can you apply your knowledge to the real life example of Alcohol Dehydrogenase and the Alcohol Flush Syndrome?