Jonathan Khalili AP Biology Period 1 Title: Enzyme Catalysis Abstract: The objective of this lab is to observe the conversion

of hydrogen peroxide to water and oxygen gas by the enzyme Catalase, as well as to measure the amount of oxygen generated and calculate the rate of the enzyme-catalyzed reaction. The methods used are titration, effect of temperature on an enzyme, and the effect of the environment on an enzyme. The results that I derived from my experiment show that the amount of H2O2 consumed not only depends on the temperature of its surrounding, but also the environment that it lives in. In conclusion, both temperature and the environment play a major role in the amount of H2O2 consumed. Introduction: This lab deals primarily with enzymes, their reaction rates, and their effects. The definition of an enzyme is a protein that catalyses (increase the rates of) chemical reactions. The molecules at the beginning of the process are called substrates, which are then converted into the products of the chemical reaction. Enzymes generally are able to catalyze more than one reaction before dying out, making enough time for new enzymes to take their place and help the next chain of chemical reactions. The effects of enzymes on chemical reactions depend on the temperature and the pH of the reaction. In most cases, the higher the temperature or pH in the surroundings of the specific substance, and the substance itself, the faster and more productive the chemical reaction can be. The colder the temperature or pH, the slower and less effective the reaction will be. While chemical reactions in cold areas have lower activation energies, it is much harder to achieve the actual chemical reaction. The protein structure of enzymes also has an effect on the chemical process. Proteins have a primary structure which is responsible for the order and sequencing of amino acids, a secondary structure which is responsible for the proteins overall conformation, a tertiary structure that is responsible for the interactions/bonds that are in the protein, as well as the threedimensional structure of the protein, and finally the quaternary structure that results from the interactions between and among several polypeptide chains. Materials: See Lab Notebook Pages 1-2 Results: Part A: 1) What is the enzyme in this reaction? Catalase 2) What is the substrate in this reaction? H2O2 3) What is the product in this reaction? O2 4) How could you show that the gas evolved is O2? Small, Subtle Bubbles.

Jonathan Khalili AP Biology Period 1 Part B: Base Line Calculation (Table 2B) Final reading of burette .5 mL Initial reading of burette 4 mL Base Line (Initial Final) 3.5 mL Part C: Uncatalyzed H2O2 decomposition (Table 2C) Final reading of burette 5 mL Initial reading of burette 10 mL Amount of KMnO4 titrant 5 mL H2O2 Spontaneously decomposed (baseline-KMnO4) -15% mL Percent of H2O2 that spontaneously decomposes (baseline-24 hours/baseline * 100) -328.57% mL

Part D: My Data (Table 2.1.1 Red) KMnO4 (ml) Time (seconds) 10 30 60 90 120 Base Line 3.5 Final Reading 10 Initial Reading 10.1 KMnO4 Used .1 H2O2 Used 3.4 3.5 4.9 10 .1

180

360 3.5 10.8 10.9 .1 3.4

3.5 3.5 3.5 3.5 11 11.1 11 10.9 27 11.9 11.1 11 16 .8 .1 .1 3.4 3.4

3.4 -12.5 2.7

Mr. Whitsons Data: (Table 2.1.2 Blue) KMnO4 (ml) Time (seconds) 10 30 60 120 Base Line Final Reading Initial Reading KMnO4 Used H2O2 Used 3.4 8.6 6.1 2.5 .9 3.4 3.4 3.4

180 3.4

360 3.4 11.6 11.6 1 Drop 3.4

10.2 11.2 8.6 1.6 1.8 10.2 1.8 2.4

11.5 11.6 11.2 11.5 .3 3.1 .1 3.3

Amount of H2O2 4 2 0 -2 -4 -6 -8 -10 -12

Jonathan Khalili AP Biology Period 1 Graph 2.1 (Amount of H2O2 decomposition)

Analysis Questions: 1. Determine the initial rate of the reaction and the rates between each of the time points. Record the rates in the table below. From the formula described earlier, recall that rate = ^y/^x.
Time Intervals (Seconds) Rates 0 to 10 .34 10 to 30 .17 30 to 60 -.416[con] 60 to 90 .09 90 to 120 .113[con] 120 to 180 .056[con] 180 to 360 .018[con]

2. When is the rate the highest? Explain why. The rate is the highest at the beginning of the reaction because the least amount of time has elapsed for the chemical reaction and the chemical reaction is still relatively unused. 3. When is the rate the lowest? For what reasons is the rate low? The rate is supposed to be the lowest at the end of the reaction because the substances are completely used up and the most time has elapsed. (Mines isnt because I did it incorrectly) 4. Explain the inhibiting effect of sulfuric acid on the function of the Catalase. Relate this to enzyme structure and chemistry. The inhibiting effect of sulfuric acid on the function of the Catalase is because enzymes work most effectively at a pH of 7.0. Sulfuric acid causes the pH to become lower, therefore deactivating the enzyme. This change can completely denature the protein.

Jonathan Khalili AP Biology Period 1 5. Predict the effect that lowering the temperature would have on the rate of enzyme activity. Explain your prediction. The effect that lowering the activation energy would have is that it would decrease the enzyme activity. This is because when the temperature is lowered, the activation energy is lowered but it is harder for the enzyme to achieve the level of energy needed to start or accelerate a new reaction. 6. Design a controlled experiment to test the effect of varying pH, temperature, or enzyme concentration. A good experiment would be bacteria. Bacteria are found in all sorts of climates and temperatures, therefore they all have different reaction rates and times. For example, a bacteria living in a hot springs area would have a higher enzyme rate and reaction rate, as well as having more reactions occur spontaneously, then say a bacteria living in the arctic, which would need more energy to make a reaction, but a lower reaction rate. Discussion: This lab was all over the place, as I had made numerous mistakes. My results are not anything like the results we are supposed to compare to. I had one negative amount of H2O2 that was consumed in the reaction, which isnt possible, and I didnt no why this happened until I found out that halfway through the lab, I realized that I titrated the same 5mL sample over and over again, therefore throwing off my results and giving me bad data. The reason for my incorrect results that I have was that I didnt read the directions more than once, as when I read it the first time, I thought I understood it and proceeded with it continuously and did not change the sample at all. A solution for the next lab we do would be to obviously read the directions over and repeatedly for every part of the lab.

Jonathan Khalili AP Biology Period 1

KEY: Red- My Data Blue- Mr. Whitsons Data

10 30 60 90 Time Elapsed (Seconds)

120

180

360