Enzymes In Action

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Lizet Enriquez B3 Biology Ap

Enzymes in Action Abstract The labs purpose is to teach you how to measure the effects of changes in temperature, enzyme concentration and substrate concentration and how these affect the reactions of enzymes in a controlled experiment. Enzymes are in our bodies and this is being shown in the experiment with the liver reacting in the hydrogen peroxide. The methods in this experiment must be followed as directed. The results show what the hypothesis had stated. It is supporting the hypothesis by showing that the factors used in the lab are affecting the enzyme-substrate reaction. This lab is teaching you that enzymes are in living things even in humans. It can denature by temperature, pH, and enzyme and substrate concentration. Time also has a great effect on a reaction in an enzymatic solution. This is shown in the experiment.

Introduction Enzymes are proteins that are produced by living cell that act as biochemical catalysts. As everybody knows an enzyme must bind to a substrate at the active site in order to make a product of the reaction formed. There are several ways an enzyme action can be affected. Some of the factors are salt concentration, pH, temperature, and if there are any activations or inhibitors acting on the enzyme. Many of these factors can make an enzyme denature or slow down making the reaction inactive. The purpose of this lab is to seeing how enzyme catalysis works. If you put

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Lizet Enriquez B3 Biology Ap

the catalase solution in different things such as hydrogen peroxide mixed with water then you are going to see different reactions occurring. Materials and Methods For the first test you will need: Hydrogen peroxide, 3 medicine cups, catalase solution, 1 test tube, test tube rack, cube of liver, 12mL syringe, 1 mL pipette, pencil and paper. For the second test you will need: Hydrogen peroxide, 8 medicine cups, 6mL syringe, 1 mL pipette, 1 mL of distilled water in medicine cups, Sulfuric acid, 12mL syringe, and Potassium permanganate. Goggles and gloves must be used in the lab at all times. Test of Catalase Activity 1 1. Transfer 10mL of Hydrogen Peroxide into a medicine cup 2. Then add 1 mL of catalase solution 3. After observing the reaction and recording the results clean all materials and dry

them Test of Catalase Activity 2 1. Put 5 mL of catalase solution into a test tube 2. After that place the test tube in the water bath for 5 minutes 3. Put 10 mL hydrogen peroxide into a medicine cup

4. Take 1 mL of the cooled boiled catalase solution and add it to the medicine cup 5. After recording the results clean all materials and dry them Test of Catalase Activity 3 1. Cut 1 cm of a liver and macerate it

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Lizet Enriquez B3 Biology Ap

2. Put it in a medicine cup containing 10 mL of hydrogen peroxide 3. After recording the results clean all materials and dry them Base Line Test 1 1. Put 10 mL of hydrogen peroxide in a medicine cup 2. Add 1 mL of distilled water 3. Then add 10 mL of sulfuric acid 4. Mix the solution well 5. Remove a 5 mL sample 6. Put the sample into a medicine cup 7. Place the medicine cup under a piece of white paper 8. Using the syringe add 5 mL of potassium permanganate one drop at a time after each

drop gently swirl and keep adding drops until you get a pink or brown color in the solution. 9. Record the results Results Test of catalase activity 1 Results Enzyme in reaction

Substrate in reaction

Product in reaction

Observation

Catalase solution

Hydrogen Peroxide

Oxygen

The

presence

of

bubbles

For the test of catalase activity 2 the solution didn’t bubble as much as activity 1 because the boiling denatures the catalase. The test of catalase activity 2 it is bubbling more than activity 1 or

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Lizet Enriquez B3 Biology Ap

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2 because the liver has catalase. If you would have boiled the liver it would had denatured the liver because of the heat. Base line test Results Final reading of burette

3 mL

Initial reading of burette

5 mL

Base line

2 mL KMnO4

2c Uncatalyzed H2O2 decomposition Final reading of burette

1 mL

Initial reading of burette

5 mL

Amount of KMnO4 titrant

4 mL

Amount of H2O decomposed

Percent of H2O KMnO4 (mL)decomposed in 24Hrs A) BASE LINE B) FINAL READING C) INTIAL READING D) AMOUNT OF KmNo4 CONSUMED E) AMOUNT OF H2O2 USED

10 180 3 4.2 5 -0.8 3.8

-7

Time (seconds)

30 60 360 3 3 3 4.6 4.6 4 5 5 5

-0.4 3.4

-0.4 1 3.4 4

133%

90

120

3 4.4 5

3 4.4 5

3 4.6 5

-0.6 3.6

-0.6 3.6

-0.4 3.4

Table of enzymatic reaction

Conclusion You will see different reactions depending on what you put the enzyme and substrate through.

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Lizet Enriquez B3 Biology Ap

This is proved in the lab when you boil the catalase and you don’t and the one that is not boiled is more reactive than the one that was boiled. This is showing how temperature affects an enzyme. Many things you do in the experiment can affect the outcome. Some of these factors are when you put the drops in the solution you must make sure you are stirring gently, you must wash the materials correctly in order to avoid left over solution affecting the outcome of the experiment.