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Enzyme Lab Finished

This lab tested how the enzyme peroxidase was affected by different factors. In Procedure 1, adding the enzyme to the substrate solution caused the solution to change color over time, showing the enzyme's activity. Procedure 2 showed that higher pH levels led to greater color changes, meaning enzyme activity increased with higher pH. Procedure 3 found that cold, room, and hot temperatures all allowed color changes, but boiling denatured the enzyme and stopped its activity.

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3K views

Enzyme Lab Finished

This lab tested how the enzyme peroxidase was affected by different factors. In Procedure 1, adding the enzyme to the substrate solution caused the solution to change color over time, showing the enzyme's activity. Procedure 2 showed that higher pH levels led to greater color changes, meaning enzyme activity increased with higher pH. Procedure 3 found that cold, room, and hot temperatures all allowed color changes, but boiling denatured the enzyme and stopped its activity.

Uploaded by

jenna727
Copyright
© Attribution Non-Commercial (BY-NC)
Available Formats
Download as DOC, PDF, TXT or read online on Scribd
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Enzyme Lab This lab tested how enzymes affected a substrate.

Objective: The purpose of this lab was to test our hypothesis on how the enzyme affected the substrate at different increments of time.

Procedure #1 Hypothesis: If the enzyme solution is added to the substrate solution, then the solution will change color. Materials:

Turnip peroxidase 0.1% hydrogen peroxide Guaiacol Distilled water 3 test tubes Stop Watch Pipettes Graduated cylinder Syringes Test tube rack Parafilm Procedure: Using the one of the three test tubes: Label the test tube substrate add 7 mL of distilled water add 0.3 mL of 0.1% hydrogen peroxide, add0.2 mL of Guaiacol 2. Cover the test tube with the parafilm and gently mix. 3. Place the test tube in the test tube rack. 4. Repeat steps 1-3 with another test tube. Label this test tube control 4. Using the last test tube: Label the test tube enzyme add 6.0 mL of distilled water add 1.5 mL of peroxidase 5. Cover the test tube with the parafilm and gently mix. 6. Place the test tube in the test tube rack. 7. Pour the substrate test tube into the enzyme test tube. 8. Immediately begin timing the reaction. 9. Rotate the test tubes and record the changes in color at 0, 1, 2, 3, 4, and 5 minutes. 10. Using the color chart figure out what the change in color was at each time increment. Independent variable: Enzyme Dependent variable: The color of the solution Control: control test tube with only the substrate Constant: Substrate

Data: Test-tube Enzyme & Substrate Control (Substrate alone)

0 min 0 0

1 min 1 0

2 min 1 0

3 min 2 0

4 min 2 0

5 min 3 0

Data Analysis: The test tube with the enzyme and substrate continued the further change color as the time increased. On the other hand the control test tube with just the substrate did not change color and remained clear over the given time period. Other groups in the class received similar results. You measured the color change at different times. Which time will you use for your later assays? Why? I will use the 3 minute color level 2 for the later procedures because this was the first color change. When you use this assay to assess factors that change enzyme activity, which components of the assay will you change? Which will you keep constant? I will change the pH of the solution and keep the enzyme constant. Conclusion: The hypothesis was supported by the experiment. The procedure used was efficient for the hypothesis tested. Any errors may have resulted from poor measuring of the enzyme and substrate.

Procedure #2 What do you predict will occur if the pH in the reaction changes? How do you justify your predictions? I think by changing the pH in the reaction changes, I think the reaction time will change. Hypothesis: If the pH is lower then the color change will be less. Materials: Turnip peroxidase 0.1% hydrogen peroxide Guaiacol Buffers with range of pH Distilled (deionized) water 12 test tubes and test tube rack Time

Pipettes Graduated cylinder Procedure: Make six sets of pairs of substrate and enzyme tubes, using a different pH substrate for each test tube For each substrate tube in a pair add: add 7 mL of distilled water 0.3 mL of hydrogen peroxide 0.2 mL Guaiacol For each enzyme test tube add: 6.0 mL and a specific pH solution 1.5 mL of peroxidase Cover each test tube with parafilm and mix Combine the substrate and the enzyme tubes for all six pairs Cover with parafilm and gently mix Begin timing the reaction Record the observation for each test tube, and record the color change at 0 minutes and at 3 minutes Independent variable: pH Dependent variable: The color of the solution Control: Control from procedure 1 Constant: Substrate and enzyme

Data: Test tube number 1 (pH 3) 2 (pH 5) 3 (pH 6) 4 (pH 7) 5 (pH 8) 6 (pH 10) Control Data Analysis:

Color after 3 minutes 1 2 2 3 3 green 3

The higher the pH was the bigger the color changes were. Other groups in the class had similar results. In procedure 1, what was the limiting factor of your baseline reaction the enzyme or the substrate? How could you modify the procedure you learned to answer this question? The enzyme; the procedure could be modify to test just the substrate after a time period. What are three or four factors that vary in the environment in which organisms live? Which of those factors do you think could affect enzyme activity? How would you modify your basic assay to test your hypothesis? Temperature, pH, air quality. Temperature and pH can affect enzyme activity. Procedures 2 and 3 test this hypothesis. Conclusion: In conclusion our hypothesis was not supported. The lower the pH did not result in a smaller color change. The technique used many have been flawed, or errors could have resulted from poor measurement and poor timing. Procedure #3 : Individual Investigation Hypothesis: If the temperature of the solution is changed, then the enzyme will respond differently at higher and lower temperatures. Materials: 8 test tubes Hot plate Ice Beakers Turnip peroxidase 0.1% hydrogen peroxide Guaiacol Pipettes Parafilm Timer Procedure: Create enzyme test tubes following the steps in procedure 1 Create substrate test tubes following the steps in procedure 1 Pour one enzyme test tube into each substrate test tube Using one of the test tube place it in a beaker of ice Record the color change after 3 minutes Using another test tube measuring the reaction after 3 minutes (room temperature)

Using the third test tube place it in a beaker of hot water First warm the water on the hot plate Record the color change after 3 minutes Using the last test tube place it in a beaker of boiling water Boil the water in a beaker on the hot plate Record the color change after 3 minutes Independent variable: Temperature Dependent variable: The color of the solution Control: room temperature test tube Constant: Substrate and enzyme Data: Test Cold (5 C) Room Temperature (24 C) Hot (50 C) Boiling (94 C)

At beginning 0 0 0 0

After 3 minutes 4 4 4 0

Data Analysis:

The experiment results show that the cold, room temperature, and hot solutions all had the same color reaction. The boiling solution did not have a color change, most likely because the enzyme denatured. The other groups in the class did not perform the same procedure, so there are no other results to compare to. Conclusion:

In conclusion, the hypothesis tested was supported. The different temperatures produced different results in not all, but one temperature. This shows temperature does affect enzyme activity. Any flaws or sources of error in the technique used to test the hypothesis may have resulted from poor measurement of the enzyme and not timing the reaction correctly.

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