AP Biology

LAB REPORT: Two Enzyme Catalysis. Group Members: Patrick Perez, Anthony Curatolo, Seung Cha, David Tseng, David Musheyev, and Alexander Coma. Date Submitted: October 28, 2011

Before the lab is completed there is a couple of background information you should be aware of.Patrick Perez AP Biology Ms. What we also observed how much oxygen was generated by the addition of enzyme to the catalyzed reaction. Rodriguez Brown Patrick Perez Introduction & Background: In the lab Two Enzyme Catalysis what we did was observe the conversions of Hydrogen Peroxide into water and oxygen gas with the addition of an enzyme. the most important being that the addition of an enzyme to a reaction will increase the rate of a .

The reason behind the increased rate of the reaction is that enzymes lower the activation energy of the reaction making the reaction occur at a much faster rate. Other factors that affect enzymes include enzyme concentration. Enzymes are . The name for this is enzyme specificity. In living organisms almost all of the crucial reactions occur within the cells. substrate concentration. This is because on the pH scale. activators. Enzymes usually function at their best and full potential at or near the pH of 7. Temperature also affects the rate of a reaction. Meaning it s not too acidic or too basic. In these reactions including reactions substrates is the name given to the targeted molecules by the enzymes.reaction. 7 is neutral. Enzymes are highly specific meaning that they only catalyze one kind of reaction. PH is one of the factors that affect enzymes. There are also factors that affect the way an enzyme performs and functions during the reaction. In biology enzymes play a crucial role. salinity. This is because without enzymes many reactions would not be able to occur within the cells. and inhibitors. Too much heat can damage enzymes.

My null hypothesis: The addition of an enzyme will have no effect on the rate of the reaction. but there are also a few things that they don t do. Enzymes do a lot. I want to be able to experience firsthand what enzymes are able to do. and are unaffected by the reaction.responsible for increased reaction rates. . From all of this background knowledge what I would like to learn from this lab is how much an addition of an enzyme to a reaction can change a reaction. My hypothesis is that I think that the addition of an enzyme will enhance the reaction making it occur faster as long as there aren t any facors that can affect the enzyme present. formation of temporary enzyme-substrate complexes. They don t change the reaction and they don t make reactions that would not occur naturally.

Materials: 2A Materials: o o o Glass Beakers Dropper H2O2 .

o o o o Cold Catalase Potato Boiled Catalase Liver 2B Materials: o o o o o o o Burette Glass Beakers H2O H2SO4 KMnO4 Syringe H2O2 2C Materials: o o o o o Glass Beakers KMnO4 Burette H2O2 H2SO4 2D Materials: o o o o o o Stop Watch to record time Catalase Glass Beakers H2O2 KMnO4 Burette Procedure: .

Check for difference from the boiled catalase. (Should be foaming) Transfer 10mL of H2O2 using a dropper into a beaker. Notice the rate of the reaction with catalase. Add 1mL of H2O Add 10mL of H2SO4 into beaker Mix solution well Remove 5mL of solution Place into another beaker . Macerate the liver Take the macerated liver and add it into the beaker with H2O2 Observe the reaction (Should be foaming) o o o o o o Add 10mL of H2O2 using a dropper into a beaker. Add 1mL of cold catalase using a dropper into the beaker which has H2O2 Mixing solution Observe reaction.2A: o o o o o o o o Transfer 10mL of H2O2 using a dropper into the beaker. Part II of 2A (Liver & Potato) [Living Tissue] o o o o o o o o 2B: Transfer 10mL of H2O2 using dropper into a beaker Macerate the bits of potatoes Take the macerated potato and add it into the beaker with H2O2 Observe the reaction. Add 1mL of boiled catalase using a dropper into the glass beaker with H2O2 Mixing the solution Observe reaction. Transfer 10mL of H2O2 using a dropper into the beaker.

My group was 180. Exercise 2A Questions: . All groups had a different time.o o o 2C: o o o o o o o o o o o 2D: o o o o o o o Place the sample under a burette containing KMnO4 Allow the KMnO4 to drop into the solution until it turns pink or brown Record information Add 15mL H2O2 in a beaker Store it uncovered for 24 hours After 24 hours has passed add 1mL of H2O Add 10mL of H2SO4 into beaker Mix well Remove a 5mL sample using syringe Place into another beaker Now place a piece of white paper over the beaker Place the sample under a burette containing KMnO4 Allow the KMnO4 it drop into sample until it turns pink or brown Record information 180 Seconds is the number of time you should observe the solution Add 10mL of H2O2 to beakers Add 1mL of catalase to the beaker Swirl solution Use a stop watch to time 180 seconds When 180 seconds is up add 10mL of H2SO4 Gather the other group s data.

2.2 mL 24Hr: 3.6mL KMnO4 Exercise 2C Questions: Uncatalyzed H2O2 decomposition Final reading of burette 13. Exercise 2B Questions: Base line Calculations Final reading of burette 17mL Initial reading of burette 13. This is because temperatures affect enzymes.2mL Initial reading of burette 10. 3. resulting in bubbles.1. The reaction did not occur because the boiled catalase did not work under boiled conditions. How could you show that gas evolved is O2? Oxygen is trying to escape. A.7 mL 24Hr: 17. What is the product in this reaction? O2 & H2O D. The catalase would not be able to work under boiled condtions.5 mL 24Hr: 13. What is the enzyme in this reaction? Catalase B.5 mL Amount of H2O2 sponatneously decomposed (mL baseline mL KMnO4) 3.5mL Base Line (Final Initial) 3. The reaction bubbled.5 mL . What is the substrate in this reaction? H2O2 C.7 mL Amount of KMnO4 titrant 3.

1 .3 1drop 3.2 11.2 1.375% Exercise 2D Questions: KMnO4 (mL0) 10 a) Base Line b) Final Reading c) Initial Reading d) Amount KMnO4 Consumed (B minus C) e) Amount H2O2 Used (A minus D) 360 3.4 4.2 11.1 2.4 180 3.4 11.4 3.6 6. For this graph you will need to determine the following: a. The independent variable: The time (Use this to label the horizontal (x) axis.5 11.5 .) Analysis of Results: 1.6 11.8 10.) b. .What percent of the H2O2 spontaneously decomposes in 24 hours? [(mL baseline mL 24hours)/ mL baseline] x 100. -9.4 8.5 11.4 120 3.0 2. The rate of the reaction is the slope of the linear portion of the curve ( y / x).6 1.2 11. The dependent variable: The amount of H2O2 Used (Use this to label the vertical (y) axis. Graph the data for enzyme catalyzed H2O2 decomposition.3 3.2 8. Data Table below shows the initial rate of the reaction and the rates between each of the time points.1 3.9 30 3.6 .4 10.6 1.4 Time (seconds) 60 90 3.

The effect of lowering the temperature on the rate of enzyme activity will make the reaction slow down. They need to add different enzymes under different conditions include different pH and temperature. This is because the reactions are much quicker in the beginning because of the enzymes. The rate is at its highest when it is first added from which is 0 to 10 seconds. It can cause the enzyme to be damaged.06 0. The protein is changed and can no longer interact with the hydrogen peroxide. The reaction is at its lowest at the end of the reaction. Therefore it is slowing back down to its original state. H2O2 and the catalase react when first combined due to its high concentration and they meet at active sites faster. This is because the reaction has already occurred and is ending. 4. They should all be seen under different conditions. There should be beakers with mixed solutions. 3. 6. The speed of the reaction will become slower. When the pH drops below a range (around 7) the enzyme doesn t function like before. pH is one of the factors that affects enzymes.The Rate of the Reaction Time Intervals (seconds) Initial 0 to Rates 10 0. 5.04 0. This is because temperature affects the enzyme.02583 2.009444 0. . They should all have the same solution. The effect of the sulfuric acid on catalase function is that it denatures the enzyme.09 10 to 30 30 to 60 60 to 90 90 to 120 120 to 180 180 to 360 0.

We are able to see that proteins are one of the most important factors in the function of enzymes. We were able to see how certain factors affected the enzymes. Factors such as inhibitors and temperature were present in this lab.Conclusion: The purpose of the lab was to show the effect of enzymes (catalase) in reactions. but we all managed to see what enzymes can do in a reaction. . My hypothesis was pretty accurate. It allows you to see how enzymes can help speed up reactions in biology. and in living organisms. The only issue was that we didn t have enough time to be able to see the effect of the enzymes. I feel like everything was rushed. As for the accuracy of the information and data it was pretty accurate.

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