Enzymes Lab Report

Instructions: In this lesson, we briefly discussed how cyclin-kinase complexes can malfunction
due to mutations to their genetic coding. In this lab activity, you will investigate the direct effects
of environmental factors on enzymes. How could things like fever, hyperthermia, changes in
blood pH, or the concentration of the enzyme affect how these protein complexes function in the
molecular control system of cell division? You will select two of these three factors to test during
your experiment. Submit your lab report to your instructor when completed.

Title:

Enzymes Lab Report

Objective(s):

Investigate the direct effects of environmental factors on enzymes.

Hypothesis:

As described above, you will be testing the effects of temperature, concentration, or pH on

enzyme activity. Select two of these factors for your investigation and create a hypothesis for
each. Remember, each part of your experiment will have a different independent variable.
Review the procedures section. Then write a prediction for each part of your investigation. Be
sure to compare your independent variable to your dependent variable using an "if /then” format
for each hypothesis statement.

Test A Hypothesis: If the concentration of the enzyme increases, then the rate of the reaction will
increase. The rate of the reaction can be determined by the amount of bubble formation as
oxygen gas is released.

Test C Hypothesis: If the pH level of the solution increases, then the reaction rate will increase.

Lab Safety:

 Cover your work surface with newspaper or other covering to protect from spills.
 Protect your eyes using goggles or protective glasses
 If sodium bicarbonate or vinegar solution gets in the eye or on the skin, flush with water.
 Oven mitt or glove for handling hot liquids should be used.

Procedure:
A list of materials and summary of steps is provided for you. You should select the procedures
that match the factor you are testing. You only need to test two factors, not all three.

Materials:

 Pick one of the following: potatoes, kiwi, peaches, cherries, apricots, bananas,
watermelon, or pineapple
 Baking soda (if testing pH)
 Water (and a way to warm it if testing temperature)
 Ice cubes (if testing temperature)
 Vinegar (if testing pH)
 Hydrogen peroxide (can be purchased at a grocery store)
 Glasses, cups, small bowls, or jars
 Something to crush fruit or veggie
 Large containers to hold hot and cold water (if testing temperature)
 Measuring cups
 Marker, paper, and tape for labeling
 Timer

Summary of Steps:

Procedures for All Tests:

1. Catalase breaks down hydrogen peroxide into water and oxygen. The oxygen will
appear as bubbles in solution. For each test, allow the reaction to complete in ALL
test tubes before recording results.
2. Rank the test tubes in by the reactivity scale below and record your results in the data
section.

REACTIVITY SCALE

0 = no reaction (no foam or bubbling)

1 = minor reaction (very few bubbles or very slow to bubble, delayed reaction)

2 = moderate reaction (medium amount of bubbles, medium speed, delayed reaction)

 3 = major reaction (a lot of bubbles, bubbles quickly and immediately)

Test A: Enzyme Concentration

1. Set up three glasses, cups, small bowls, or jars. Label each with a number.

2. Cut fruit or potato into cubes. Place one cube into glass 1.

3. Cut a second cube of fruit or potato into four smaller pieces. Put these pieces into
glass 2.

4. Take third cube of fruit or potato and crush it down with the mortar and pestle or
another utensil used for crushing. Please the crushed fruit or potato in glass 3.

5. Pour ¼ cup of hydrogen peroxide into each glass.

6. Using your timer and the reactivity scale provided above, estimate the rate of
reaction in each glass. Record your results in Table 1.

7. Clean the glasses for the next test.

Test B: Temperature

1. Add one cube of fruit or potato to glasses 1, 2, and 3.

2. Boil water (100⁰) and carefully place this water in a large container to create a hot
water bath. Place glass 1 in the hot water bath for five minutes.

3. Add ice to a second container to create an ice water bath (0⁰C). Place glass 3 in the
ice water bath for five minutes.

4. Glass 2 will remain at room temperature, which is approximately 25⁰C.

5. After five minutes, using protective glove or oven mitt, remove the glasses from
the ice and hot water baths.

6. Pour ¼ cup of hydrogen peroxide into each glass.

7. Using your timer and the reactivity scale provided above, estimate the rate of
reaction in each glass. Record your results in Table 2.

8. Clean the glasses for the next test.

Test C: pH level

1. In glass 1, mix equal amounts of baking soda with water. Add ¼ cup of hydrogen
peroxide into glass 1. This is your basic solution with a pH above 7.
 2. In glass 3, mix equal amounts of vinegar with water. Add ¼ cup of hydrogen
peroxide into glass 3. This is your acidic solution with a pH below 7.

3. In glass 2, add ¼ cup of hydrogen peroxide, then add water to equal the volume of
the solutions in glass 1 and 3. This is your neutral solution with a pH around 7.

4. Add one cube of fruit or potato to glasses 1, 2, and 3.

5. Using your timer and the reactivity scale provided above, estimate the rate of
reaction in each glass. Record your results in Table 3.

6. Clean the glasses for the next test.

Variables:

You selected two tests for this investigation. List and explain your controlled variables,
independent variable, and dependent variable for each part of the investigation.

Remember, controlled variables are factors that remain the same throughout the experiment. An
independent (test) variable changes so that the experimenter can see the effect on other variables.
The dependent (outcome) variable will change in response to the test variable.

Test A

Controlled variables: Temperature, pH level, amount of hydrogen peroxide, kind of fruit

and the length of the experiment.

Independent variable: Concentration of the enzyme.

Dependent variable: Formation of bubbles as oxygen gas is released.

Test C

Controlled variables: Temperature, amount of hydrogen peroxide, kind of fruit, and the
length of the experiment.

Independent variable: The pH level.

Dependent variable: Formation of bubbles as oxygen gas is released.

Data:

Complete the tables below that correspond to the tests you selected:
Table 1- Enzyme Concentration
Glass Contents and Observations Reactivity Value

1 (large cube) Hardly any bubbles formed. 1

2 ( four small cubes) Bubbles formed slowly. 2

3 (mashed cubes) Bubbles formed the most in this 3

glass.

Table 2
Glass Contents and Observations Reactivity Value

Table 3- pH Level

Glass Contents and Observations Reactivity Value

1.( 1 cube in acidic solution) Hardly any bubbles formed 1

2.( 1 cube in neutral solution) Bubbles formed slowly. 2

3. ( 1 cube in basic solution) Bubbles formed the most in this 3

basic solution.

Conclusion:

Write a conclusion statement that addresses the following questions:

 How did your experimental factors affect enzyme behavior in both tests? Explain your
results.
In Test A, the enzyme catalase was more accessible to react with the hydrogen peroxide
in the smaller cubes and the smashed cube. This explains why the reactivity scale was
higher in these two cases.
The enzyme catalase has an optimum pH level of 9, which is considered basic. This
explains the results of Test C, where we observed no reaction in the acidic and neutral
solutions and a major reaction in the basic solution.
 Does your data support or fail to support your hypotheses?
The experimental data clearly supports the hypotheses.
 Discuss any possible sources of error that could have impacted the results of this
investigation.
 A possible source of error could be the method by which the solutions were prepared in
Test C.

Post-lab Reflection Questions

Answer the reflection questions using what you have learned from the lesson and your
experimental data. It will be helpful to refer to your class notes. Answer questions in complete
sentences.

1. What kind of scenarios could change the temperature, pH, or enzyme concentration level
in the human body? (Describe at least two.)

During periods of strenuous exercise, core body temperature elevates rapidly. This is due
to the fact that only a small fraction of the energy produced is actually used for muscle
contraction, the rest is lost as heat and dissipated throughout the body, thereby increasing
body temperature.
A normal blood pH ranges from 7.35 to 7.45. Blood acidosis occurs when the blood pH
drops below 7.35. When carbon dioxide reacts with water in the blood, it forms carbonic
acid, making the blood acidic. Respiratory acidosis is a medical condition whereby the
lungs are unable to expel carbon dioxide from the body quickly enough.

Predict the two possible outcomes if temperature or pH were changed within the
molecular control system of cell division.

Cell division is regulated by internal checkpoints using multiple mechanisms. The most
important cell cycle regulators are the cyclin-dependent kinases ( CDKs). Kinases, like
all enzymes, have optimal conditions under which they can function. If these regulators
don’t function properly, an organism may end up with too few or too many cells. This
can cause varying outcomes- from harmless hair loss to cancerous tumors.

Samples from Streptococcus cultures are tested to determine the rate of cell division
under different conditions. The average results of the tests performed are presented in the
graphs below:
 a) Predict what would happen to this species of bacteria if it infected a human body
with a body temperature of 98.6 ⁰F and a pH range of 7.35 - 7.45.
The Streptococcus cultures would probably proliferate rapidly.

b) Provide reasoning to support the claim that a medication that lowers fever could
increase cell division in this bacterium.

The optimal temperature for growth in Streptococcus bacteria is around 98.6

degrees, which is considered a normal human body temperature. A medication
that lowers fever sets the body temperature back to 98.6 degrees, which is the
optimum temperature for proliferation in these bacteria.

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