Courtney Zahn

Bio 1315L
November 8, 2013
Lab Report 3

How Enzymes Function

Part 1: Catalase Activity
Purpose: Test controls without the different effects of enzyme activities (Controls)
Hypothesis: Catalase and hydrogen peroxide will be the positive control. Catalase and
sucrose solution will be the negative control (no specific reaction). Water and Hydrogen
peroxide will have not cause an increase in the height of the bubble column.
Materials:

3 test tubes
Catalase buffered at pH 7.0
Hydrogen peroxide
Water
Ruler
Sucrose

Procedures:
Use wax pencil to label three clean test tubes at the 1cm and 5cm mark.
Test Tube 1
Fill to the first mark with catalase buffered at pH 7.0. Fill to the second mark with
hydrogen peroxide. Swirl well and wait 20 seconds for bubbles to develop.
Measure the height of the bubble column, and record your results.
Test Tube 2
Fill to the first mark with water. Fill to the second mark with hydrogen peroxide.
Swirl well to mix and wait at least 20 seconds. Measure the height of the bubble
column, and record your results.
Test Tube 3
Fill to the first mark with catalase. Fill to the second mark with sucrose solution.
Swirl well to mix; wait 20 seconds. Measure the height of the bubble column, and
record your results.
Variables:
Independent Variable: contents in test tubes
Dependent Variable: height of bubble column
Control group: water and hydrogen peroxide
Experimental group: catalase and hydrogen peroxide/catalase and sucrose solution.

Results:
Test Tube
1

Contents
catalase hydrogen
Peroxide
water hydrogen
peroxide
catalase sucrose
solution

2
3

Bubble Column Height

Explanation

5 mm

(+) Control

none

control group

2 mm

(+) Control

Height of Bubble Column in mm

Test Tube 3

Height of Bubble
Column in mm

Test Tube 2
Test Tube 1
0

Conclusion:
Test tube 1, which contained Catalase and hydrogen peroxide, had the highest
bubble column, just as we hypothesized. Test tube 1 is the positive control group. Test
tube 3 should have had no reaction instead of the 2mm bubble column, because Catalase
A manual error might have occurred. Maybe hydrogen peroxide was put instead of
sucrose solution. Test tube 2 showed no increase in height of the bubble column and is
the control group.

Part 2: Effect of Temperature

Purpose: Determine how temperature affects enzyme activity.
Hypothesis: Increase of temperatures will increase enzyme activities.
Materials:

3 Test tubes
Ruler
Catalase buffered at pH 7.0
Hydrogen peroxide
Refrigerator
Incubator
Boiling point

Procedure:
Label and mark three clean test tubes at the 1cm and 5cm levels.
1. Fill each tube to the first mark with catalase buffer pH 7.0.
2. Place tube one in refrigerator, place tube 2 in an incubator, and tube 3 in a boiling
water. Wait 15 minutes.
3. Remove test tubes and fill hydrogen peroxide to the 5cm mark.
4. Swirl well and wait 20 seconds
5. Measure the height of the bubbles and record results
Variables:
Independent Variable: temperatures
Dependent Variable: Bubble height column
Control: Contents and time
Results:
Test Tube
Temperature
1 Refrigerator
10C
2 Incubator
37C
3 Boiling water
80C

Bubble column height

1mm
3mm
none

Explanation
partial increase
increase reactions
denatured

Height of bubble column in mm

3 Boiling Water (80 degrees C)
Height of bubble column in
mm

2 Incubator (37 degrees C)

1 Refrigerator (10 degrees C)

0

0.5

1.5

2.5

3.5

Conclusion:
Increase of temperature does increase enzyme activity, but if the temperature is
too hot, the protein will denature and will not be able to react.

Part 3: Effect of Concentration on Enzyme Activity

Purpose: Determine how concentration affects enzyme activity.
Hypothesis: Increase amount of concentration of enzymes will increase enzyme activity.
Materials:
3 Test tubes
Ruler
Catalase buffered at pH 7.0
Hydrogen peroxide
Water
Procedure:
Test Tube 1
Mark to 1cm and 5cm. Fill the first mark with water and the second mark with
hydrogen peroxide. Swirl and wait 20 seconds. Measure the height of the bubble
column and record the results.
Test Tube 2
Mark to 1 cm and 5cm. Fill to the first mark with buffered catalase and to the second
mark with hydrogen peroxide. Swirl and wait 20 seconds. Measure the height of the
bubble column and record the results.
Test Tube 3
Mark to 3cm and 7cm. Fill to the first mark with buffered catalase and to the
second mark with hydrogen peroxide. Swirl and wait 20 seconds. Measure the
height of the bubble column and record the results.
Variables:
Independent Variable: concentration
Dependent Variable: bubble column height
Results:
Tube
1
2
3

Amount of Enzyme
none
1cm
3cm

Bubble height column

none
5mm
2mm

Explanation
no enzyme
more
little more

Height of Bubble Column in mm

Height of Bubble Column
in mm
0

Conclusion:
The results of this experiment are not consistent with the hypothesis. There might
have been a manual error because test tube 3 should have had the highest bubble column.
Part 4: Effect of pH on Enzyme Activity
Purpose: Determine how pH affects enzyme activity.
Hypothesis: Every enzyme has an optimal pH at which its rate of reaction is highest.
Materials:

3 Test tubes
Ruler
Hydrogen peroxide
Water adjusted to different (pH)s
Non buffered catalase

Procedure:
1. Label and mark three clean test tubes at the 1cm, 3cm, and 7cm levels. Fill each
tube to the 1cm level with non-buffered catalase.
2. Tube 1- Fill to the second mark with what adjusted to pH 3. Wait 1 min. fill to the
third mark with hydrogen peroxide. Swirl to mix and wait 20 seconds. Measure
the height of the bubble column and record the results.
3. Tube2- Fill to the second mark with water adjusted to pH 7. Wait one min. Fill the
third mark with hydrogen peroxide. Swirl to mix and wait 20 seconds. Measure
the height of the bubble column and record.
4. Tube 3- fill to the second mark with water adjusted to pH 11. Wait one min. fill
the third mark with hydrogen peroxide. Swirl to mix, and wait 20 seconds.
Measure the height of the bubble column and record results.
Variables:
Independent Variable: pH
Dependent Variable: bubble height column
Control: hydrogen peroxide
Results:
Tube
1
2
3

pH
3
7
11

Bubble height column

0
38mm
30mm

Explanation
acidic
increase in activity
lost activity

Height of Bubble Column in mm

Test Tube 3 (pH 11)

Height of Bubble Column

in mm

Test Tube 2 (pH 7)

Test Tube 1 (pH 3)
0

10 15 20 25 30 35 40

Conclusion:
Results show that the enzyme meets its optimal activity between pH 7 and pH 11, and the
enzyme begins to decrease. We used the potato enzyme for the pH experiment and the
commercial enzyme for the first 3 experiments. Our results would be more conclusive if
we used the same enzyme for each experiment. The results were better with the potato
enzyme (bubble columns were higher).

References:
Mader, Sylvia S. Chapter 1. Biology. 11th ed. New York: Mcgraw Hill, 2013. 1-8. Print.