1. Wednesday, 9/12/12 Finish Toothpickase Activity  must be completed today!!! – Due tomorrow!!!!! 1 st lab this week – set up today, lab Thursday & Friday – no open toed shoes! Chp.4,5,8 Test next Monday  TUESDAY! – Carbon, Biomolecules, Enzymes

2. Use small spiral notebook to set up and record labs Use the first page to create a table of contents # the notebook pages

3. AP Bio Lab # 13 Enzyme Activity Pg. 3 – Start with Lab Title

4. Before doing this lab, you 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 (view graph) That changes in temperature, pH, enzyme concentration, and substrate concentration can affect the initial reaction rates of enzyme-catalyzed reactions (view graph)

5. Under title, subtitle first section: Learning Objectives Students will use the catalase reaction to demonstrate different variables’ effect on enzyme activity: – such as temperature and the presence of catalase. Students will observe the reaction of hydrogen peroxide and catalase: – establish a baseline determining the amount of hydrogen peroxide in a 1.5% solution, – Determine the rate of spontaneous conversion of hydrogen peroxide to water and oxygen – determine the rate of hydrogen peroxide decomposition by enzyme catalysis.

6. Next section, title: Background Info In this experiment, we will be using the enzyme catalase. – found inside peroxisomes in almost all organisms. – catalyses the breakdown of hydrogen peroxide to water and oxygen. – Without this enzyme, hydrogen peroxide would destroy our cells because it is a toxic metabolic waste product of aerobic respiration. 2 H 2 O 2  2 H 2 O + O 2 (gas)

7. Background Info Cont: Reaction is spontaneous (will occur naturally), but very slowly. If catalase is boiled, it will become denatured and will no longer function to break down hydrogen peroxide. The rate of this reaction will decrease as the concentration of hydrogen peroxide (the substrate) decreases.

8. Safety Read all instructions before beginning lab. Wear personal protective eyewear (PPE) at all times, included during clean-up. Wear gloves and aprons, Potassium Permanganate will stain clothes! Be careful with chemicals, Sulfuric Acid is highly corrosive! Wash hands before leaving lab.

9. Set-up picture

10. Part A: Demo Mix Hydrogen Peroxide (H 2 O 2 ) + Catalase

11. The General Procedure H 2 SO 4 stops the reaction! Measure using Potassium Permanganate Drops

12. Part B: Establishing a Baseline – Determining the Amount of Hydrogen Peroxide (H 2 O 2 ) in a 1.5% solution Add Hydrogen Peroxide (H 2 O 2 ) Add Water (control) Add Sulfuric Acid (H 2 SO 4 ) Titrate with Potassium Permanganate (KMnO 4 ) Amount of (KMnO 4 ) used to titrate is proportional to amount of (H 2 O 2 ) present in the solution

13. Part C: Demo Overnight Control Group: Do NOT add any catalase enzyme, and measure rate of Hydrogen Peroxide Spontaneous Decomposition

14. Part D: Rate of Hydrogen Peroxide Decomposition by Enzyme Catalysis 1.After you get your baseline 2.Add Hydrogen Peroxide (H 2 O 2 ) and Catalase solution, swirl for 10 seconds 3.Add Sulfuric Acid (H 2 SO 4 ) to stop reaction 4.Titrate with Potassium Permanganate (KMnO 4 ) 5.Repeat for 30, 60, 120, & 180 seconds

17. 1. Start Adding one drop at a time 2. Swirl, if solution turns back clear, continue adding one drop at a time 3. Stop adding drops when solution remains a pinkish – yellowish color, measure & record amount of KMnO 4 used.

18. Next section, title: Data Glue in all 4 data tables – Be sure to keep the titles attached so you know what data table is used for which section of the lab

19. Next section, title: Conclusion You will receive conclusion questions to glue in and answer in this section