Jennifer Putman AP Bio 2nd Enzyme Catalysis

Introduction: This labs purpose was to understand biochemistry through the chemical processes of enzymes. In order to understand the lab, one must learn the functions, activities, and rates of enzymes. After completing the lab, one must be able to measure the effects of temperature change, pH, enzyme concentration, and substrates in a controlled experiment. Necessary vocabulary includes:
enzyme- proteins produced by living cells that lower activation energy for chemical reactions substrate- substance acted upon by the enzyme reaction active site- place where enzyme binds substrate during reaction assay- test to measure the proteins biological activity after a reaction catalase- common enzyme found in nearly all living organisms that are exposed to oxygen baseline- control for assays

Abstract: In Exercise 2A, three trials are tested using hydrogen peroxide in a beaker. Each trial, a small amount of catalase is added (1-normal, 2-boiled, or 3-living tissue) to determine the decomposition of hydrogen peroxide to oxygen gas. In Exercise 2B, the baseline of hydrogen peroxide is established by adding distilled water to peroxide, and finally adding sulfuric acid to stop the reaction. A 5mL sample is removed and placed in a beaker where drops of potassium permanganate until pink, where the baseline can be determined as how much peroxide is remaining. In the time-course determination, six cups of hydrogen peroxide were mixed with catalase for a finite time to react. Sulfuric acid stopped the reaction so that an assay could be taken through titration of potassium permanganate. In Exercise 2D, the procedure of 2B was repeated, just using peroxide instead of water. For the main portion of the lab, the time trials, the independent variable is the amount of time (seconds) the reaction between the hydrogen peroxide and the catalase was allowed to react (10, 30, 60, 120, 180, 360). The dependent variable is the amount of hydrogen peroxide that remains after the reaction is stopped. The control is the baseline determined prior to the start of the time trials. If the catalase is allowed to react longer with the hydrogen peroxide, more oxygen gas will become a product of the reaction. Materials & Methods: Catalase is used as the enzyme because it is common in many living organisms that are exposed to oxygen, where it catalyzes the decomposition of hydrogen peroxide to oxygen and water. Catalase has one of the highest turnover numbers of all enzymes, which allows faster lab completion due to rapid decomposition. Hydrogen peroxide, in turn, is used because it decomposes quickly with catalase. Sulfuric acids purpose is to quickly stop the reaction between the catalase and hydrogen peroxide by lowering the pH. Enzymes decompose at pH levels much higher or lower than 7. Potassium permanganate is titrated into the stopped

Jennifer Putman AP Bio 2nd reaction to determine the amount of evaporated oxygen gas, or how much hydrogen peroxide did not react. In this lab, syringes were used instead of burets due to lack of quantity, and ease of lab completion. Data:

Jennifer Putman AP Bio 2nd Discussion: Catalase increases the decomposition rate of hydrogen peroxide. As is started in the hypothesis, the longer the catalase is allowed to react with hydrogen peroxide, the less that is left post-reaction. This lab shows how catalase added to hydrogen peroxide leads to the release of oxygen, boiled catalase is denatured, and the presence of catalase in living things can lead to the breaking down of hydrogen peroxide in the body. The natural decomposition hydrogen peroxide becomes slower than decomposition with the addition of catalase. If hydrogen peroxide was to decompose naturally, life could not survive on the slow reaction rate. Possible errors are present in the lab. The lab was conducted over three days, using a different mixture of catalase that could have been slightly off from the previous day. Also, the time trial assays were taken on two consecutive days and, even though the containers of stopped reactions were tightly covered, the air is extremely dry, resulting in possible overnight evaporation. Questions: 1) Why must a baseline constantly be determined for each section? A baseline ensures that the enzyme was prepared correctly, and allow a control example for when completing the time trials. 2) Why couldnt other enzymes be used? Catalase is the most abundant and easily obtainable enzyme that serves the purpose of hydrogen peroxide decomposition. 3) How does catalase work in humans (purpose)? True catalase purpose is not clear, but in some, a catalase deficiency can result in Type II diabetes. Some people have limited catalase and have no negative effects. Peroxiredoxin is a more common antioxidant enzyme-attacker of hydrogen peroxide in animals than catalase.