1.

Enzyme Catalysis

Kyle Dunn

10/16/2021
Instructions:
1. Please read all of the introduction and background information within the investigative
manual.
a. Once you have done so answer the prelab question BEFORE completing any of
the lab’s activities.
2. Once you have completed the prelab questions proceed to the activities of the lab within
the investigative manual.
a. As you read through the instructions for completing each activity make sure you
also:
a.i. Complete any instructions (append photos, etc)/ and answer any questions
found in the post lab questions for each activity.
a.ii. Take the photos of your experiments in each activity as directed below.
IMPORTANT: Don’t clean-up your lab until you know what portion of
the experiment you need to take a picture of.
3. Here is a video that will introduce you to the lab and its main concepts. The student is
encouraged to watch it.
a. Enzyme Catalysis

Prelab Questions
1. Define activation energy. Will activation energy for a reaction be higher or lower when
an enzyme is present to catalyze it? Why?

Activation energy describes the amount of energy needed for a reaction to begin (Clark et al.,
2018). The activation energy when an enzyme catalyzes for a reaction will be lower, as
enzymes are more efficient at catalyzing due to their ability to continually repeat their
function without consuming themselves or being destroyed in the process.

2. There are a number of conditions that will impact the rate an enzyme is capable of
catalyzing a reaction including: temperature, pH, enzyme concentration, and substrate
concentration. In this lab you will investigate the effects of temperature and enzyme
concentration on the rate of an enzyme. How though will substrate concentration affect
the rate of an enzyme? To investigate this, google “How does substrate concentration
affect enzyme activity?” and answer the questions below:

a. Does an increase in substrate concentration lead to an increase in enzyme activity?

Yes, increasing substrate concentration also increases the rate of reaction to a certain point.
(OpenStax, 2015)

2 © 2016 Carolina Biological Supply Company

 a. Are there limitations to the answer you gave in “a.”? In other words, can an
increase in substrate concentration ever NOT lead to a detectable increase in
activity of the enzyme? Why or why not?

Yes. There is a limit to the increase in activity that can be experienced in substrate
concentration. Once a significant limit is reached, the substrate concentration produces no
significant change in the reaction rate (Enzyme Activity, 2021).

3. As stated above, in this lab we will be investigating the effect of enzyme concentration
(activity 1) and temperature (activity 2) on the rate of an enzyme. BEFORE you begin
each activity please fill out the purpose and hypothesis statements found below.

Activity 1
Instructions:
1. Open the investigative manual. Locate all the needed materials supplied in the kit
and those you will need to supply yourself.
2. Lay them out in your work area.
3. Read through the entire set of instructions found in the investigative manual for the
activity to avoid making mistakes when you go to execute the experiment.
4. Once you have read through the instructions go back to step 1 and begin executing
the experiment.
5. Please answer the questions below and/or append appropriate representations of data
(photos, graphs, etc). REMEMBER don’t clean up until you have taken the
appropriate photos of your experiment as described below.

Purpose statement: (This should be the question the experiment is attempting to address. It
should be written as a question.)

How will the enzyme catalase effect the catalyzation and decomposition of hydrogen peroxide
(H2O2) at various concentrations? How does the concentration affect the decomposition rate of
the substrate hydrogen peroxide?

Hypothesis statement: (This should be an “if/then” testable prediction that addresses the
question/purpose of the lab.)

If small pieces of filter paper which have been soaked in various concentrations of catalase are
exposed to H2O2, then those which have been exposed to a higher concentration of catalase will
catalyze at a faster rate, resulting in the released oxygen gas bubbles carrying these pieces to the
surface at a faster rate than those with less exposure.

3 © 2016 Carolina Biological Supply Company

Evidence/Claim statement: (This should be a statement regarding whether your hypothesis was
supported or refuted and what data/evidence allows you to make this claim.)

The data supports the hypothesis in this instance. We can see that as a piece of filter paper is
exposed to a higher concentration of catalase, it will rise to the top of the H202 mixture at a
faster rate. This is because the catalase present on the piece of filter paper is producing oxygen as
it decomposes the surrounding H202, which lifts the paper to the surface of the mixture. Thus,
the faster a piece of filter paper rises to the top, the faster the catalase is catalyzing.

Reflection statement: (This should be a statement of what you learned, how your understanding
changed, if you have new questions, and what connections can you make between the lab and the
content in the book and other assignments.)

The biggest takeaway from this experiment is the efficiency of enzymes, in my eyes. Witnessing
first hand the catalyzing process due to the introduction of enzymes demonstrated how
efficiently and quickly they can work.

Photo 1 – Activity 1

Take a picture and insert the image(s) of your medicine cup after step 8. of the “Enzyme
Concentration” section in activity 1 of the investigative manual:

4 © 2016 Carolina Biological Supply Company

Data Table 1

0% 25% 50% 75% 100%

Depth of
H2O2 0 0 2.3 2.3 2.3
Solution

5 © 2016 Carolina Biological Supply Company

 0% 25% 50% 75% 100%
(From
step 11.)

Trial 1
180sec 180sec 148sec 112sec 68sec
Time

Trial 2
180sec 180sec 144sec 111sec 72sec
Time

Avg.
180sec 180sec 146sec 111.5sec 70sec
Time

Rate
(d/t)
0cm/s 0cm/s 0.016cm/s 0.02cm/s 0.032cm/s
(From
step 12.)

Graph 1 – Activity 1

Insert your graph from step 13. of the “Enzyme Concentration” section in activity 1 of the
investigative manual:

6 © 2016 Carolina Biological Supply Company

 1. Having now completed activity 1 and your graph go back and fill in your Evidence/Claim
and Reflection statements and then answer the question(s) below.

2. Based on the information in Data Table 1 and your graph, explain the relationship
between percent concentration of catalase and reaction rate. From this what can we conclude
about the effect that enzyme concentration has on the rate of a reaction?

As the percent of catalase increases, so too does the reaction rate. We can thus conclude that the
higher the enzyme concentration, the higher the rate of reaction.

7 © 2016 Carolina Biological Supply Company

 3. Using the graph, predict the rate if catalase concentrations are 10%, 60% and 85%.

I would predict 10% to still have no reaction rate, as 25% did not yield any reaction in my
experiment. At 60%, I would predict approximately 0.018 cm/sec as a reaction rate, as this is
roughly halfway between the two rates of reaction at 50% and 75%. While 60 is closer to 50 than
75, this predicated reaction rate would likely be, on average, about 0.018 cm/sec or 0.0175
cm/sec. 85% would perhaps be close to 0.025 cm/sec, for similar reasons to what I just listed for
60%, though with the numbers swapped for 75% and 100%. We begin to see a significant
increase in the rate of reaction around this time, so we could expect it to begin to grow quickly at
this point.

Activity 2
Instructions:
1. Open the investigative manual. Locate all the needed materials supplied in the kit
and those you will need to supply yourself.
2. Lay them out in your work area.
3. Read through the entire set of instructions found in the investigative manual for the
activity to avoid making mistakes when you go to execute the experiment.
4. Once you have read through the instructions go back to step 1 and begin executing
the experiment.
5. Please answer the questions below and/or append appropriate representations of data
(photos, graphs, etc). REMEMBER don’t clean up until you have taken the
appropriate photos of your experiment as described below.

Purpose statement: (This should be the question the experiment is attempting to address. It
should be written as a question.)

How will temperature impact the decomposition rate of hydrogen peroxide when catalase is
exposed to it as a result of enzyme activity?

Hypothesis statement: (This should be an “if/then” testable prediction that addresses the
question/purpose of the lab.)

If catalase soaked paper filter paper is exposed to an H2O2 solution at varying degrees of
temperature, then the warmer the temperature, the faster the decomposition rate. Inversely, as the
temperature lowers, the decomposition rate will take a longer amount of time.

Evidence/Claim statement: (This should be a statement regarding whether your hypothesis was
supported or refuted and what data/evidence allows you to make this claim.)

8 © 2016 Carolina Biological Supply Company

 My hypothesis was not supported by the evidence. Rather, the data showed that as temperature
increased, there would come a point where the rate of decomposition would decrease before
seemingly falling off completely. My hypothesis was correct that a decrease in temperature
would show a slower reaction rate, but an increase also appears to have the same effects.

Reflection statement: (This should be a statement of what you learned, how your understanding
changed, if you have new questions, and what connections can you make between the lab and the
content in the book and other assignments.)

It makes sense that at a certain temperature, it would become too hot for the enzyme to function
correctly. I believe I was thinking that a much higher temperature would need to be achieved
before this would occur, but it appears that even very subtle changes in temperature can greatly
impact the rate at which enzymes catalyze.

Data Table 2

10 deg. below Room temp. 10 deg. Above 20 deg. Above 30 deg. Above
13oC 23oC 33oC 43oC 53oC
Depth of
H2O2 2.25 2.25 2.25 0 0
Solution

Time 154sec 104sec 137sec 180sec 180sec

Rate (d/t) 0.015cm/sec 0.022cm/sec 0.016cm/sec 0cm/sec 0cm/sec

Graph 1 – Activity 2

Insert your graph from step 17. of the “Effect of Temperature” section in activity 2 of the
investigative manual:

9 © 2016 Carolina Biological Supply Company

 1. Having now completed activity 2 and your graph go back and fill in your Evidence/Claim
and Reflection statements and then answer the question(s) below.

2. Based on the information in Data Table 2 and your graph, Is there an optimum
temperature for catalase that makes it the most productive in terms of reaction rate? At what
temperature does the rate decrease? If your data does not show a decrease do you think there
is a temperature in which rate would begin to decrease? Explain why.

I believe more experiments with more precise temperatures would need to be conducted to
find the most productive temperature. However, at 43 degrees C, it was apparent that the rate
had already begun to slow. Room temperature may be ideal, although it is possible a
temperature slightly higher than this may yield faster results. The reaction rate will also
decrease as the temperatures falls as well, so I'm lead to believe room temperature once again
may be ideal.

References

10 © 2016 Carolina Biological Supply Company

Enzyme Activity. (2021, July 31). https://chem.libretexts.org/@go/page/16022
Clark, M. A., Jung, C., & Douglas, M. (2018). Biology 2e. Houston: OpenStax.
OpenStax, Biology. OpenStax CNX. (May 13, 2015) http://cnx.org/contents/185cbf87-c72e-
48f5-b51e-f14f21b5eabd@9.85.

11 © 2016 Carolina Biological Supply Company