Effect of pH on the enzyme catalase

Aim: To investigate the effect of pH on the activity of the enzyme catalase which is present in the liver. Equipment list: 40 ml of hydrogen peroxide (H2O2) 1 mol L-1 hydrochloric acid in dropper bottle 1 mol L-1 sodium hydroxide in dropper bottle 2 mol L-1 hydrochloric acid in dropper bottle 2 mol L-1 sodium hydroxide in dropper bottle Liver Safety risks: Distilled water 6 test tubes Cutting board Safety gloves (latex) and glasses Electronic balance Stop watch Scalpel 10ml measuring cylinder Universal indicator and pH strips

Hydrochloric acid, sodium hydroxide and hydrogen peroxide are corrosive to skin and can cause burns and serious damage if spilt into eyes. To avoid this wear safety glasses. The liver may have some disease germs that can get inside the body and cause illness. Wear safety gloves and wash your hands after the practical to avoid this. The scalpel is very sharp and it can cut through skin so be cautious when using it.

Variables: Independent- the pH concentration Dependent- the activity of the enzyme (catalase) Controlled variable- the hydrogen peroxide ml in each test tube Control- Test tube 1

Method: 1. Using a measuring cylinder measure 5ml of hydrogen peroxide and pour it into each of the six test tubes which are labelled 1-6. 2. Do not add any chemicals to test tube 1 and label this as the control. 3. Using a pH strip and a pH chart determine the pH of the solution in test tube 1. 4. Add 5 drops of hydrochloric acid into test tube 2 and then 8 drops into test tube 3. 5. Add 10 drops of distilled water into test tube 4 and then 5 drops of 1mol L-1 sodium hydroxide into test tube 5 and 5 drops of 2mol L-1 of sodium hydroxide into test tube 6. 6. Using universal indicator strips and the pH chart determine and record the pH of solution in each test tube from 1-6. 7. Place all test tubes in a water bath and maintain the temperature at 37oC. 8. Wear safety gloves and safety glasses to avoid touching an acid or alkali. 9. Using a scalpel, cutting board and electronic balance measure and cut six pieces of liver of mass 3g each. 10. Carefully add one piece of liver into test tube 1 and using a stopwatch measure the time taken by oxygen gas bubbles to reach the top of the test tube. Thet Win HSC Bio Page 1

Effect of pH on the enzyme catalase

11. 12. 13. 14. Record the time taken using an appropriate method. Repeat steps 10-11 using the solution in test tubes 2-6. Using a thermometer record the temperature at which the reaction occurred. Repeat the experiment 10 times.

Results/observations: Test tube Chemicals in test tube pH Temperature Time taken (oC) by gas to reach the top (s) TRIAL 1 37 12 37 105 Time taken by gas to reach the top (s) TRIAL 2 18 95 Average time taken (s) 15 100

1 2

Hydrogen peroxide ONLY 1 mol L-1 hydrochloric acid + hydrogen peroxide 2 mol L-1 hydrochloric acid + hydrogen peroxide Distilled water + hydrogen peroxide 1 mol L-1 sodium hydroxide + hydrogen peroxide 2 mol L-1 sodium hydroxide + hydrogen peroxide

4.5 2

37

135

165

150

37

14

10

12

37

120

80

100

13

37

160

140

150

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Effect of pH on the enzyme catalase

Conclusion: The trend observed is a bell shaped curve whereby the activity of the enzyme is greatest around a pH of 6-7. This is because the enzyme catalase is found in the liver inside the human body and it requires a neutral i.e. a pH of 7 for optimal metabolic efficiency and function. The experimental test tubes were also placed in a water bath with a maintained temperature of 37oC to ensure similar conditions as to what are found inside the body. This experiment was valid for a number of reasons. One such was that the aim was appropriately tested. Also the particular results obtained were useful information as it justified the activity of enzymes at different pH concentrations. In terms of reliability it was quite reliable as the results were reproduced consistently and the mean was taken. The reaction which occurred was the decomposition of hydrogen peroxide into hydrogen and oxygen with the catalase enzyme.

2H2O2(aq)

2H2O(l) + 02(g)

Essentially pH concentration lower or higher than 6/7 results in a slower rate of reaction as the enzymes dont work as efficiently. At higher temperatures they die i.e. become denatured and can no longer function.

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