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Potato Enzyme Lab

INTRODUCTION
An enzyme is a protein that speeds up or slows down a specific chemical reaction in an organism. A good rule of thumb
is to remember that enzyme names end in -ase. This will help in identifying enzymes in further readings. Generally
enzymes are catalysts.
Hydrogen peroxide is a toxic chemical that is produced in many organisms during metabolism. Organisms must get rid
of this toxin to survive. One reaction turns the hydrogen peroxide into water and oxygen. The enzyme that helps with
this reaction is called catalase. This is found in both plants and animals. In this lab we will use potatoes as our catalase
source. The reaction equation is:
Catalase
2H
2
O
2
2H
2
O + O
2


Additional Information that May Help: "Hydrogen peroxide bubbles when it comes into contact with an enzyme
called catalase. Most cells in the body contain catalase, so when the tissue is damaged the enzyme is
released and becomes available to react with the peroxide. Catalase allows hydrogen peroxide (H
2
O
2
to be
broken down into water (H
2
O) and oxygen (O
2
). The bubbles you see when you pour oxygen on a cut are
bubbles of oxygen gas. Blood, cells, and some bacteria (e.g., staphylococcus) contain catalase, but it is not
found on the surface of your skin so pouring peroxide on unbroken skin will not cause bubbles to form. Also,
because it is so reactive, hydrogen peroxide has a shelf life once it has been opened, so if you don't see
bubbles form when peroxide is applied to an infected wound or bloody cut, there is a chance your peroxide
is no longer active." http://chemistry.about.com/od/medicalhealth/f/Why-Does-Hydrogen-Peroxide-Bubble-On-A-Wound.htm
Background:
What is an enzyme? A catalyst? A chemical reaction? A metabolism?
What is hydrogen peroxide?
Where is catalase found?
When is catalase released?
What are the bubbles evidence of? What are the bubbles made of?
What can happen to the enzymes when you heat/cook potatoes, plants, animals?
PURPOSE
1. Observe the breakdown of hydrogen peroxide toxin by potatos enzyme catalase.
2. Determine factors that influence how quickly the reaction takes place.
3. Determine factors that influence how well enzymes function.
4. Use graphic analysis (graphing) to analyze our results.
(Use points 1-3 above to write your own purpose)
PRELAB QUESTIONS
1. Read the purpose and the procedure. After having read these what is the dependent and independent variable
in the experiment?
Independent Variable: State of the potato (diced, ground, cooked)
Dependent Variable: Reaction time/speed, amount of bubbles.

2. What test tube is the control group? Why?
The test tube with the sand because it is not living and does not contain an enzyme in it.
3. Formulate an If/then hypothesis. Remember if/then hypothesis always follow the same form of if the
________________(dependent variable) is related to the _______________(independent variable) in ________ way, then given
_____(condition) I predict ______________(outcome).
If the reaction time is related to the state of the potato, the amount of bubbles will be an indicator of the enzyme
catalyzing the hydrogen peroxide break down.
If the potato is cooked, then there will be no reaction because the enzymes were denatured when the potato was heated.
If the potato is diced, then there will be a reaction; it will be slower because there is less surface area and less tissue
broken compared to the ground potato.
If the potato is ground, then there will be a significant reaction because many tissue cells in the potato were broken and
there is greater surface area for the chemical reaction with hydrogen peroxide to break down.
4. What is the point of adding sand to test tube one?
To ensure that hydrogen peroxide doesnt react with the container it is in and doesnt react with non living things.
Materials: 4 test tubes, test tube rack, graduated cylinder, hydrogen peroxide, potato (ground, diced, diced and
cooked), sand, timer (You can copy this exactly)
PROCEDURE (You can copy this exactly- attach your comic strip)
1. Mark test tubes 1-4.
2. Fill each test tube with 5mL hydrogen peroxide.
3. Make initial observations of test tube one.
4. Add a pinch of sand to test tube one.
5. Observe and record observations.
6. Obtain about 1g ground potato. To grind the potato use a mortar and pestle and a pinch of sand. Record the
exact mass and add it to test tube 2.
7. Time how long it takes for the reaction to take place. Youll know the reaction is taking place because bubbles
are made.
8. Obtain about 1g of diced potato. Record the exact mass and add it to test tube 3.
9. Time how long it takes for the reaction to take place. Youll know the reaction is taking place because bubbles
are made.
10. Obtain about 1g of cooked potato. Record the exact mass and add it to test tube 4.
11. Time how long it takes for the reaction to take place. Youll know the reaction is taking place because bubbles
are made.
DATA AND OBSERVATIONS (Copy this exact table into the lab)
Test tube Observations Time for reaction (s)
1.







2.







3.







4.









DATA ANALYSIS
1. Calculate a rate of reaction per gram of potato for each trial. Rate = time/grams
2. Graph the data. Include a title, labels, units, and a key.
Type of graph: bar/column graph
Conclusion: use complete sentences. Each of the following answers MUST include the word because
This begins the analysis section of the lab report.
1. Summarize and explain the results.
You can use your own data or another groups data, but explain what is shown in the graph and tables.
2. Did you support or refute your hypothesis? Why? What data leads you to that conclusion?
The evidence shows.
The results indicate.
From the data it can be concluded
3. What did the catalase do? How do you know?
The catalase triggered the chemical reaction and breakdown of hydrogen peroxide into water and oxygen. The
bubbles were evidence as well as the products of the chemical reaction.

4. What did grinding the potato up do to the rate? Why did this change the rate?
Grinding the potato caused the chemical reaction to begin- the tissue of the potato was broken and the enzymes
could reach and react with hydrogen peroxide. Grinding the potato also increased the surface area and therefore
the rate of the chemical reaction.
5. What did cooking the potato do to the rate? What conclusion can you draw about that result?
Cooking the potato slowed down the chemical reaction because the enzyme and other proteins that catalyze
this reaction were denatured- or killed, destroyed, broken apart, not viable, dont work as they should.

Potential Errors: (Analysis Section)
Cooked potato not cooked enough
Errors in timing depending on recognizing bubble formation
Started timer only when you saw the first bubbles; started timing when potato was put in and stopped timing when we
saw bubbles.
Old hydrogen peroxide (already separated into water and oxygen)
Potato not ground well- didnt break enough tissue to release enzyme

Conclusion:
Hypothesis proven or not (1 sentence)
Future experiments dealing with hydrogen peroxide and catalase in living organisms (plants, animals, bacteria)



Part B- pH
Each enzyme has an optimal pH.
A change in pH can alter the ionization of the R groups of the amino acids. When the charges on the
amino acids change, hydrogen bonding within the protein molecule change and the molecule changes
shape. The new shape may not be effective.
The diagram below shows that pepsin functions best in an acid environment. This makes sense because
pepsin is an enzyme that is normally found in the stomach where the pH is low due to the presence of
hydrochloric acid. Trypsin is found in the duodenum, and therefore, its optimum pH is in the neutral
range to match the pH of the duodenum.

Most cells form hydrogen peroxide (H
2
O
2
) as a waste product of aerobic respiration. Hydrogen
peroxide is toxic and must be converted to water and oxygen by the enzyme catalase.

Hydrogen peroxide is also commonly used as a household disinfectant. It bubbles when it is applied to
cuts and scrapes because catalase is present in the fluids of the broken cells. As the equation above
shows, the bubbles are oxygen gas (O
2
).
In the experiment below, bubbling will be used as an indication that a reaction is occurring. A faster
reaction will have more bubbling.
B1. Predict how pH will affect the rate of reaction in the three different situations described below and
write your hypothesis or hypotheses on the answer sheet. At this point, you do not know if your
hypothesis is true or false. It is acceptable to create a hypothesis which will be shown to be false.
B2. Mark three test tubes with a wax pencil 3 cm from the bottom and 6 cm from the bottom.
B3. Add 3 cm of hydrogen peroxide to each tube.
B4. Fill one of the tubes to the 6 cm mark with 5 m HCl.
B5. Fill another tube to the 6 cm mark with 5 m NaOH.
B6. Fill the third tube to the 6 cm mark with distilled water.
B7. Cut three pieces of potato that are one cubic centimeter (cc) each.
B8. Crush each of the three pieces of potato separately with a mortar and pestle and add one crushed
piece to each of the three tubes that you prepared in steps 2-6 above. It may be helpful to use a small
spatula when transferring the potato to the tube.
B9. Write your observation of the amount of bubbling in table 2. We are only interested in the amount
of bubbling. We are not interested in any change in color or whether the potato floats or sinks.
B10. Use pH paper to measure the pH of each tube and record your measurements in table 2 on the
answer sheet.
B11. At what pH did catalase function best in this experiment?
B12. Do your results support your hypothesis? Explain.
B13. In part A of this lab, you worked with rennin, an enzyme found in the stomach of mammals. The
pH of the stomach is normally about 2.0. At what pH do you predict rennin works best?
B14. What can you say about pH and enzyme functioning? Is there a single pH that enzymes function
best at or does it depend on the enzyme?