Enzyme (Lactase) Inquiry

Group Members names: Nico Hartojo, Marcus Cassar, Jake Christensen

Problem: Does temperature affect the effectiveness of the lactase enzyme.

Research Hypotheses: (include at least 2 with a prediction)

If the lactase becomes denatured at extreme temperatures, then its function

of digesting lactose will decrease, causing a greater amount of glucose in
these extreme temperatures.

Procedure:

1. Get 8 test tubes.

2. In the test tube add 2 mL of glucose solution (positive control).
3. In the second tube add 2 mL of whole milk (control).
4. In the third tube add 2 mL of fat free milk (control).
5. In the fourth tube add 1 mL of lactase (negative control).
6. In the fifth tube add 1 mL of lactase and 2 mL of whole milk mix
at room temperature.
7. In the sixth tube add 1 mL of lactase and 2 mL of fat free milk
mix at room temperature.
8. In the seventh tube add 1 mL of lactase and 2 mL of lactose
(positive control).
9. Let the solution sit for 3 minutes.
10. Take and record the temperature of all 8 solutions.
11. Dip paper in each solution to test for glucose.
12. Dip paper in each solution to test for protein.
13. Dip paper in each solution to test for the pH level.
14. Record the initial color of paper.
15. Let each piece of paper sit for 2 minutes.
16. Record the final color of paper.
17. Gather new test tubes with the solutions of test tubes 7
and 8.
18. Heat up each solution on a hot plate in boiling water (100
degrees C).
19. Record the temperature of solutions.
20. Test for protein, glucose, and pH.
21. Record the initial color of paper.
22. Wait for 2 minutes.
23. Record the final color of paper.
 24. Gather new test tubes with the solutions of test tubes 7
and 8.
25. Put the beakers in refrigerator for 10 minutes.
26. Place the test tubes in for 5 minutes.
27. Record the temperature of solutions.
28. Test for protein, glucose, and pH.
29. Record the initial color of paper.
30. Wait for 2 minutes.
31. Record the final color of paper.

Data: (include a table with observations and a Graph)

Lactase at Room Temp

Test Tube Contents Final Presence of Temperatur

Glucose Lactase e of
Levels Enzyme Solution

1 Glucose ++ - Room

2 Whole Milk - - Room

3 Fat Free - - Room

Milk

4 Lactase - + Room

5 Lactase and ++ + Room

Whole Milk

6 Lactase and +++ + Room

Fat Free
Milk

Lactase at 100C

Test Tube Contents Final Presence of Temperatur

Glucose Lactase e of
Levels Enzyme Solution

1 Glucose ++ - Boiling
100C
2 Whole Milk - - Boiling
100C

3 Fat Free - - Boiling

Milk 100C

4 Lactase - + Boiling
100C

5 Lactase and - + Boiling

Whole Milk 100C

6 Lactase and - + Boiling

Fat Free 100C
Milk

Lactase at 0C

Test Tube Contents Final Presence of Temperatur

Glucose Lactase e of
Levels Enzyme Solution

1 Glucose ++ - Boiling 30C

2 Whole Milk - - Boiling 30C

3 Fat Free - - Boiling 30C

Milk

4 Lactase - + Boiling 30C

5 Lactase and - + Boiling 30C

Whole Milk

6 Lactase and + + Boiling 30C

Fat Free
Milk

Conclusion/Recommendations: (Can be bulleted)

When at boiling temperature the enzyme did not break down the
lactose, and there was not any glucose. This most likely happened
because the enzyme was denatured.
 When it was cold the enzyme broke down the lactose a bit better
than the boiling. The average level of the glucose was about 2.5.
When at room temperature the enzyme broke down the lactose
very well, leaving a high amount of glucose.
The enzyme works at its best at room temperature.
If the enzyme is exposed to extreme temperatures, cold and hot,
it will denature.
Shape is key to function, and when its denatured, the lactase
cannot do its job of turning lactose into galactose and glucose.