Enzyme Catalysis

PRE-LAB QUESTIONS
1. Briefly describe three factors that could alter the activity of an enzyme.

The three factors that can alter the activity of an enzyme are temperature, salinity and
acidity. For example, if you place and enzyme solution in the refrigerator or freezer, that
would alter its activity. Adding vinegar to the enzyme solution would also alter its
activity.

2. How do you think the optimal temperature for enzymes found in thermophile bacteria
living in hot pools differ from that of the enzymes found in your body?

The optimal temperature is around 100 degrees Celsius, while the temperature of the
enzymes found in the body is around 37 degrees Celsius.

3. How do you think the optimal pH of enzymes in the stomach compares to the optimal pH
for enzymes in blood?

I would expect the optimal pH to be very low and acidic, since the stomach contains high
amounts of acid naturally. The optimal pH if the blood, I would think to be closer to
7/neutral.

4. Take a look around your house and identify household products that work by means of an
enzyme. Name the products and indicate how you know they work with an enzyme.

In my home, I found that my hand soap uses a-amylase. This enzyme breaks down the
starch-based soil to simpler forms for the detergent to remove. The laundry detergent I
use has protease and lipase. The lipase in the detergent breaks down the fats and oils.
Protease works to break down the protein chains.

Enzyme Catalysis

©eScience Labs, LLC 2018

 Enzyme Catalysis

EXPERIMENT 1: ENZYMES IN FOOD

Data Tables

Purpose (What question is this experiment designed to answer?):

Using the starch indicator, iodine-potassium iodide, will amylase be present in the food samples?

Hypothesis (Based on what you’ve learned in the pre-lab materials, write and If/Then statement
regarding the outcome of this experiment.):
If two different foods are tested against a starch and an amylase control, then the results would
show which foods have each. It is expected that both the banana and apple will contain both
since fruits naturally contain starch and amylase and both contain carbohydrates.

Table 1: Presence of Starch and Amylase

Resulting Color
Substance (where sample was Starch Present? Amylase Present?
applied)
Reddish brown No Yes
Negative Control
Deep purple Yes Yes
Starch Control
Reddish brown No Yes
Saliva
Deep purple Yes Yes
Fruit/Vegetable 1
Deep purple Yes Yes
Fruit/Vegetable 2

Claim (What are the results of your experiment?):

Both the Banana and apple contained starch and therefore contained amylase.

Evidence (What data did you collect that supports your claim? Explain the meaning behind the
data and the calculations.):
By comparing the fruits to the controls, I found that both contained starch and the enzyme
amylase. I know this because, both samples became a dark purple when using the starch
indicator. The reason they turned that color is because that is the color that shows the presence of
starch as we determined by the starch control. Since they both contain starch, they will contain
amylase. This is known by the white spot in the center of the dark purple. All starch contains
amylase in order for the starch to be broken down

Post-Lab Questions

1. What is the function of amylase? What does amylase do to starch?

©eScience Labs, LLC 2018

Enzyme Catalysis

Amylase is a digestive enzyme that acts on starch in food, breaking it down into smaller
carbohydrate molecules.

5. What were your controls for this experiment? What did they demonstrate? Why was
saliva included in this experiment?

There was a starch control that showed if the samples contained starch. The next control
was the negative control which showed if the samples contained amylase. Another
control was the saliva sample. Saliva contains amylase; therefore, it can serve as a
comparison for the enzyme. It is important to note that if a certain food doesn't contain
amylase, your saliva provides the amylase needed to start the digestive process as you
chew your food.

6. Explain how this experiment works. What type of result indicates that the sample does
contain amylase? What type of result indicates that the sample does not contain amylase?

When amylase is present the starch sprayed on the paper towel will be converted to sugar.
IKI turns starch purple. When all the starch has been converted to sugar by the enzyme
amylase you will get a white spot on the paper where the sample containing amylase was
applied, if this spot has the same purple color as the surrounding area that shows the
sample did not contain amylase.

7. Which of the foods that you tested contained amylase? Which did not? What
experimental evidence supports your claim?

Both the banana and apple had. Amylase. I know this because both samples had a white
spot on the paper as noted in the above question.

8. Did you get the expected results? Why or why not? If you were to repeat this experiment,
what would you do differently to improve the experiment?

The results of this experiment were what I had expected. My hypothesis stated that both
fruit samples would contain starch and amylase, which was correct as both samples had
dark/deep purple circles with the white amylase spot. If I were to repeat this experiment, I
would test a variety of fruits.

9. Looking at the chemical reaction provided here, propose a different method that could be
used to measure amylase activity: Starch + H2O maltose

©eScience Labs, LLC 2018

 Enzyme Catalysis

I am actually unsure of that what this question is asking.

10. In addition to amylase, there is another digestive enzyme that is secreted by the salivary
glands. Conduct outside research to determine the name of this enzyme, what substrate it
acts on, where in the body it is activated, and what it is used for. Include references to
information sources used.

Another digestive enzyme that is secreted by the salivary gland is the lysozyme enzyme.
Lysozyme is used to kill any bacteria that enters your mouth through the food you’re
eating (Becker,2019). It’s been found to provide defense against gram-positive bacteria
like Bacillus Streptococcus. It works by breaking down the polysaccharides that are
present in the cell wall and once the cell wall is broken down, the bacteria die. This
action is called lysis, the enzyme that does this is lysozyme.

11. The stomach contains enzymes that aid in digestion, including proteases which digest
proteins. Why don’t these enzymes digest the stomach and small intestine, which are
partially composed of protein?

Pepsin is secreted as pepsinogen in the stomach and converted into active pepsin by HCl
present in gastric juice. HCl and pepsin both can be very corrosive to the stomach lining.
To prevent this from happening, there is a thick layer of mucus that keeps the stomach
lining and small intestine covered.

12. Saliva does not contain amylase until babies are two months old. How could this affect an
infant’s digestion?

Babies under two months of age are only fed breast milk and or formula. The breast milk
and formula only contain simple sugars that don’t require amylase in order to be broken
down.

Reflection (Discuss what you have learned by doing this experiment. How have your ideas
changed? Do you have any new questions? What connections did you make between the lab and
lecture?):

In this experiment I learned the importance of enzymes. I had a very basic knowledge of
enzymes prior to this lab but I didn’t realize its true extent. I didn’t realize that enzymes are in
almost everything around me and the things I use or eat daily. A connection I made ding this
experiment was that enzymes are very selective. Enzyme Catalysis
1.

©eScience Labs, LLC 2018

 Enzyme Catalysis

EXPERIMENT 2: EFFECT OF TEMPERATURE ON ENZYME

ACTIVITY
Data Tables

Purpose (What question is this experiment designed to answer?):

What are the effects of temperature on enzyme activity based on the amount of oxygen
produced?

Hypothesis (Based on what you’ve learned in the pre-lab materials, write and If/Then statement
regarding the outcome of this experiment.):

If the sample of hydrogen peroxide is submerged into hot water and yeast is added, then the
volume of the balloon will increase.

Table 2: Balloon Volume vs. Temperature

Test Tube Number Temperature Final Balloon Final Balloon Final Balloon
and Condition (˚C) Length (cm) Width (cm) Volume (cm3)

1 - (Room
24 10.9 6.2 1755.08cm^3
Temperature)

2 - (Cold) 12 11 5.8 1550.02cm^3

3 - (Hot) 42 11.2 7.1 2364.96cm^3

Claim (What are the results of your experiment?):

The results of the experiment show that the greater the temp, the great the balloon volume will
be.

Evidence (What data did you collect that supports your claim? Explain the meaning behind the
data and the calculations.):

The balloon with the sample placed in the fridge had the lowest volume (1550.02 cm^3). The
balloon with the sample kept at room temperature was second (1755.08 cm^3). The balloon with
the sample placed in hot water came in highest for volume (2364.96 cm^3). Gas expands when
it’s heated that is why the warmest sampled had the largest balloon.

Post-Lab Questions

1. What is the enzyme in this reaction?

Zymase

©eScience Labs, LLC 2018

Enzyme Catalysis

13. What is the substrate in this reaction?

Glucose

14. What are the two products in this reaction? Explain how one of the products is used to
measure enzyme activity.

Ethanol and carbon dioxide were the two products. Carbon dioxide inflates the balloon
and in turn is used to measure the enzyme activity. By measuring the balloon, we are able
to see how the enzyme reacted since enzyme activity is affected by temperature.

15. Create a graph of your results, plotting balloon volume vs. temperature. Describe any
correlations you observe.

Balloon Volume vs. Temperature

2500

2000

1500

1000

500

0
24 12 Balloon Volume (cm^3) 42

16. State a detailed conclusion about the effect of temperature on this specific enzyme based
on your results.

©eScience Labs, LLC 2018

Enzyme Catalysis

If the temperature is increased, then the final balloon volume will be greater. When
working with the zymase enzyme the optimal temperature is 40 degrees Celsius.
Anything below this won't perform as well. Enzymes are very selective when it comes to
their conditions and this is the prime condition for zymase.

17. Does your result match what you had expected based on how you know enzyme activity
is affected by temperature? Explain.

1. The results in this experiment did match my hypothesis. For this particular enzyme, it
reacts better to an environment at 40 degrees Celsius or more. Sine there was a sample at
room temperature and one sample was placed in the fridge, I knew that they wouldn't
perform as well because they wouldn’t be at a high enough temperature due to the fact
that enzymes are selective when it comes to their environment.

18. What changes would you make to improve this experiment? Provide at least one
suggestion for improvement.

One thing I noticed is that in doing this experiment alone I can’t do the part of the
experiment where you take the temperature, place the balloon and measure the balloon all
at the same time. If there were more people, we could each take a test tube to work on
that way that part could be completed all at the same time. With just myself, the sample
in the fridge and the sample in the hot water stayed in their environment longer then the
sample at room temperature which could affect the results.

19. What could you do to increase enzyme activity in this experiment? Provide at least one
suggestion.

To increase enzyme activity, I would but the samples in even warmer conditions and
potentially use more yeast.

20. Do animals have an enzyme that breaks down hydrogen peroxide? How could you test
for the presence of such an enzyme?

Catalase is an enzyme animals have which is present in the liver. Catalase breaks down
hydrogen peroxide into oxygen and water. To test the presence of Catalase you can use a
sample of animal liver. Put the sample n a test tube and add hydrogen peroxide. The
reaction will create foam if the enzyme is present.

©eScience Labs, LLC 2018

 Enzyme Catalysis

21. What changes would you make to this experiment to determine the optimal temperature
for this particular enzyme? Do a web-search to find the optional temperature for this
enzyme. Include references to information sources.

Increasing the temperature of all of the samples can be done to find the optimal
temperature. 30-40 degrees Celsius is the. Optimal temp range for zymase.

Reflection (Discuss what you have learned by doing this experiment. How have your ideas
changed? Do you have any new questions? What connections did you make between the lab and
lecture?):

During this lab experiment I realized how important and impactful temperature is on enzyme
activity. Before taking this class and completing this experiment I had thought that the majority
of things performed better at room temp, which is not the case as I have learned.

Sources

Becker, A. (2019, March 02). Names of the Enzymes in the Mouth & Esophagus. Retrieved July
10, 2019, from https://sciencing.com/names-enzymes-mouth-esophagus-17242.html

Buddies, S. (2012, March 08). The Liver: Helping Enzymes Help You! Retrieved July 10, 2019,
from https://www.scientificamerican.com/article/bring-science-home-liver-helping-enzymes/

OpenStax CNX. (2018, March 28).  Biology. Retrieved

from https://openstax.org/details/books/biology-2e.

©eScience Labs, LLC 2018