Before the lab, one should understand:
•The general functions and activities of enzymes;
•The relationship between the structure and function of enzymes •The concept of initial reaction rates of enzymes;
•How the concept of free energy relates to enzyme activity; •That change in temperature, pH, enzyme concentration, and substrate concentration can affect the initial reaction rates of enzyme-catalyzed reactions; and •Catalyst, catalysis, and catalase
After the lab, one should be able to:
•Measure the effects of changes in temperature, pH, enzyme concentration, and substrate concentration on reaction rates of an enzyme-catalyzed reaction in a controlled experiment; and •Explain how environmental factors affect the rate of enzyme-catalyzed reactions.
When one mixes catalase and hydrogen peroxide with sulfuric acid for a short period of time, there is a high rate of increase for the decomposition of hydrogen peroxide to hydrogen and water. However, after one mixes these substances together for a longer period of time than necessary, substrate becomes overly-saturated in the solution which causes the enzyme not to work properly.
•Reaction vessels (cups)
First, we made the base line assay to determine the fluctuation from the normal reaction mixture without catalase. This was the index of the initial concentration of H2O2 in solution. To establish a baseline, start by putting 10 mL of 1.5% H2O2 into a clean glass beaker. Add 1 mL of H2O (instead of enzyme solution). Add 10 mL of H2 SO4 (1.0 M). Mix well. Remove a 5-mL sample. Place this 5-mL sample into another beaker and assay for the amount of H2O2 as follows. Use a syringe to add KMnO4 , a drop at a time, to the solution until a persistent pink or brown color is...