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Enzyme Kinetics

Investigate “Asian Glow” syndrome by studying Alcohol Dehydrogenase kinetics. Learn how to use a spectrophotometer, perform enzyme kinetics experiment, analyze data and understand different inhibition mechanisms.

About This Simulation

In the Enzyme Kinetics Lab, students learn how substrates are converted into products by catalysis. Additionally, students learn about the kinetics of enzyme involving the Michaelis-Menten equation and various rate constants. Content regarding DNA mutation and hyperactivity are also covered. In this lab, students run experiments using the enzyme Alcohol Dehydrogenase on a wild and mutant type to learn about Asian Glow syndrome.

Using spectrophotometer to measure enzyme reaction

Students access a fully equipped workbench where they prepare the Alcohol Dehydrogenase reaction and measure the product of Acetylaldehyde using a spectrophotometer. Students learn about the concept of spectrophotometer, how to prepare a master mix and how to calculate dilution. Students prepare a reaction in a 1 ml cuvette and measure the amount of product formed using the spectrophotometer.

Supplementary 3D animation illustrates what happens on the molecular level when the substrate and co-factor enter the active site; dialog explains the situation as product is formed. Students are also prompted with quiz questions during the 3D animation to test their understanding on concepts. In the interactive animation, students identify the substrate by clicking on the different molecules.

For every measurement, students receive a progress curve displaying amounts of product formed over time. At the end of the experiment, results are presented as an Excel spreadsheet. Students must analyze their outcome data and plot their 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, students will be able to understand the Asian Glow syndrome. With the newly added module of enzyme inhibition, students are asked to perform different enzyme inhibition experiments using three different inhibitors. To begin, Students measure product formation using several inhibitor concentrations, extract the data, create their own Lineweaver-Burk plot and solve the Ki.

We have recently upgraded the Enzyme Kinetics lab by implementing a mathematically based simulator. This provides students with a larger flexibility in conducting their experiments. Students can change parameters such as substrate concentrations, enzyme concentrations, temperature or pH and receive the corresponding results. In this semi-guided module, students may 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, students will be familiar with the kinetics of enzyme Alcohol Dehydrogenase, performing the experiment as well as analyzing the data outcome. By using a real life example of Alcohol Dehydrogenase and the Asian Glow syndrome, students will be able to understand about enzyme kinetics and relate it to their own daily life.

Screenshots

Collaborators

Prof. Karen Skriver

Prof. Karen Skriver

Department of biology

University of Copenhagen

Dr. Gert Dandanell

Dr. Gert Dandanell

Department of biology

University of Copenhagen

Dr. Lasse Kristoffer Bak

Dr. Lasse Kristoffer Bak

School of Pharmaceutical Sciences

University of Copenhagen

Dr. Michael May

Dr. Michael May

Department of Science Education

University of Copenhagen

Dr. Susanne Jacobsen

Dr. Susanne Jacobsen

Institute for System Biology

Technical University of Denmark

Dr. Maher Abou Hachem

Dr. Maher Abou Hachem

Institute for System Biology

Technical University of Denmark

Learning Objectives

  • Learning the experimental design of enzyme kinetics
  • Understanding the Michaelis-Menten model of enzyme kinetics
  • Analyzing spectrophotometer data and calculate Km and Vmax
  • Understanding that kinetics of an enzyme can be modified by genetic mutations
  • Understanding inhibition kinetics by using several types of inhibitors

Techniques In This Simulation

  • Spectrophotometry
  • Data analysis of enzyme kinetics measurements

Collaborator

Collaborator

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