Catalase Enzyme Lab

INTRODUCTION:
What would happen to your cells if they made a poisonous chemical? You might think that they would die. In
fact, your cells are always making poisonous chemicals. They do not die because your cells use enzymes to
break down these poisonous chemicals into harmless substances. Enzymes are proteins that speed up the rate
of reactions that would otherwise happen more slowly. You have hundreds of different enzymes in each of
your cells. Each of these enzymes is responsible for one particular reaction that occurs in the cell. The enzyme
is not altered by the reaction.

In this lab, you will study an enzyme that is found in the cells of many living tissues. The name of the enzyme
is catalase (KAT-uh-LAYSS); it speeds up a reaction which breaks down hydrogen peroxide (H2O2), a toxic
chemical, into 2 harmless substances--water (H2O) and oxygen (O2).

The reaction is as follows: 2H2O2 ----> 2H2O + O2

This reaction is important to cells because hydrogen peroxide (H2O2) is produced as a byproduct of many
normal cellular reactions. If the cells did NOT break down the hydrogen peroxide, they would be poisoned.

PART A - NORMAL CATALASE ACTIVITY

1) Use a transfer pipette to transfer approximately 3 mL of the 3% hydrogen peroxide solution into a clean test
tube. Add a small piece of liver to the test tube.
● Observe the bubbles; what gas is being released? O2

2) Write down notes & qualitative observations of the reaction.

● Bubbled a lot
● Liver scrunches up
● Liver deteriates

**Throughout this investigation you will estimate the rate of the reaction (how rapidly the solution
bubbles) on a scale of 0-5 (0=no reaction, 1=slow...., 5=very fast).
Assume that the reaction during this step proceeded at a rate of "4".

3) Recall that a reaction that absorbs heat is endothermic; a reaction that gives off heat is exothermic.
● After the initial reaction, did the test tube get warmer or colder? warmer

● Is the reaction endothermic or exothermic? exothermic

PART B - IS CATALASE REUSABLE?
4) Pour the liquid in the test tube off into a second clean test tube. Assume the reaction is complete.

● What is this liquid composed of? Oxygen & Hydrogen

5) Add another piece of liver to this 2nd tube.

● What happened? Explain why this occurred.

Nothing happened because the oxygen inside the hydrogen peroxide already escaped and it’s only water.

6) Add another approximately 3 ml of hydrogen peroxide to the remaining liver in the ORIGINAL test tube.

* What happened when more hydrogen peroxide was added to the original liver? Explain WHY this
occurred.
It bubbled more because more hydrogen peroxide was introduced for the liver to react again. The liver acted
as catalase and broke down the hydrogen peroxide compound again.

● Are enzymes reusable? Briefly Explain.

Yes enzymes are reusable because the liver reacted again after the first reaction.

PART C – FACTORS THAT AFFECT THE ENZYME ACTIVITY (DATA)

Both temperature, pH and ionic conditions (like salts) can affect the rate of enzymatic activity. Extremes of
both conditions can alter the shape or denature the enzyme thus rendering it useless. You will investigate how
varying temperatures, ion (salt) concentrations OR pH levels surrounding the liver impacts the enzyme
catalase.
● What factor will you be observing & analyzing? (temp, ions, or pH) ions

LAB DESIGN: You will work with your group to design a lab to test your specific factor. Your goal is to test
different levels / amounts of the factor for at least 2-3 trials each. The general outline for the lab design is:

1) Place individual pieces of liver in each test tube / small beaker

2) If doing pH or ion concentration, add the specific solution to test to the test tube (should cover the liver)
3) Let liver soak in the solution for at least 5 min., remove liver from solution and place in a new separate
test tube with H2O2 to test
4) If doing temp, place liver in a test tube with some water and place in specific temp for at least 5 min.
5) Remove the liver from the water and place in a new separate test tube with H2O2 to test
6) Be sure to rate each reaction on the same 1-4 scale from Part A & B & make qualitative observations
You actual design should be more specific and include your # of trials, the specific ion concentrations, pH /
or temperatures you will test. Also include all the materials you plan to use.

If you are doing temperatures, there are hot plates, ice, etc. needed to create the ice / hot water bath to test the
temperatures you would like (and thermometers, of course - in Celsius!). Below are the solutions we will have
available for you to test, if you like. You DO NOT have to test them all.

pH - 3, 5, 7, 9, 11

Ion Concentration (salinity) - distilled water (0%), 0.5%, 1%, 3% and 5%

Materials: Procedure: (# each step & be specific; at least 2-3 trails)

● Distilled water (0%)
● 3% Ion Concentration
● 5% Ion Concentration
● 3 Clean test tubes
● 3 Beakers
● 3 Plates
● Dropper for hydrogen
peroxide
● Tweezer
● 3 Liver pieces
● Hydrogen Peroxide
● 3 Stopwatches
1. Gather Materials
2. Place a piece of liver in each test tube / small beaker
3. Add the 0%, 3%, and 5% solutions to test to a beaker each
(should cover the liver)
4. Let liver soak in each solution for at least 5 min.
5. Remove a piece of liver from each solution and place in a plate
6. Add 3mL of Hydrogen Peroxide to 3 different clean test tubes
using a dropper
7. Place a piece of liver from each solution to the new test tubes
8. Start a stopwatch for every experiment
9. Record the time of each experiment
10. Clean the 3 test tubes
11. Repeat steps 6 through 9 two more times.

Hypothesis:
The lower ionic concentration the liver was soaked in, the faster the reaction occurred
and finished/ the more effective the enzyme catalase was.

CLICK HERE to view the Google Sheets Template -- Create a single copy & share with all of your group
members. Use this to record data & create your final graph(s).
● Step 1: Refer to the corresponding table with data for your selected factor; use this to fill in the table
below. Include the factor you chose, units, and the data for ALL TRIALS.
 ● Step 2: Calculate the average of the 3 trials. You may do this “by hand” or through Google Sheets
functions (select desired cells and input “=AVG”).

● Step 3: Use Google Sheets to create ONE line graph of your data; this should illustrate ALL trials. You
may include the average if you’d like. (Be sure to include a title, labels, etc.)

● Step 4: Transfer the graph you created from Google Sheets to the space provided below the data table.
(Adjust sizing as needed.). To transfer the graph use command C to COPY and command V to PASTE
(or right click).
Data Table:

FACTOR REACTION RATES

ionic concentration TRIAL #1 TRIAL #2 TRIAL #3
(%) (Seconds) (Seconds) (Seconds) AVERAGE
0 25.9 15.03 17.6 19.51
3 30.13 24.73 26.77 27.21
5 30.35 33.36 23.25 28.99

ionic concentration AVERAGE

(%) (Seconds)
0 19.51
3 27.21
5 28.99

Qualitative Observations
● The higher concentration ion groups didn’t bubble as high compared to lower concentration ion
groups
● Lower concentration ion groups bubbled faster than higher concentration ion groups

Graph:
PART D – DATA ANALYSIS (CER)
● Use the CER process to write a 1-2 paragraph analysis of the results of the lab on the next page
● 1st → make a CLAIM of what happened in the experiment regarding your selected factor.
● 2nd → provide AT LEAST 3 pieces of evidence (data) that support your claim
○ Refer to the data in the tables & graphs you created as evidence
○ Be sure to reference SPECIFIC quantitative data as your evidence
● 3rd → explain how each piece of evidence supports the claim (this is your REASONING)
○ You are basically explaining what happened in the experiment using evidence as support!!
In the experiment, the enzyme catalase’s effectiveness was higher when
it did not absorb any ionic solutions. The enzyme catalase’s effectiveness was lowered
significantly the higher concentration ionic solutions it absorbed. It can be clearly inferred
from the “Ionic Concentration vs Average Reaction Rate in Seconds” chart, that the lower
concentration of ionic solutions the enzyme catalases absorbed, the faster their reactions
occurred and finished. For example, the test group that did not absorb any ionic solution had
the fastest/most effective reaction by resulting in 19.51 seconds compared to the 3%
concentration group that had an average reaction time of 27.21 seconds and the 5%
concentration group that had an average reaction time of 28.99 seconds. To put these pieces of
evidence in context, the faster a reaction happens and finishes, the more effective the enzyme
catalase that was present in the reaction was. Additionally, we observed that the lower
concentration ion groups bubbled faster than higher concentration ion groups. This qualitative
data that shows test groups with enzyme catalases that absorbed the lower ionic
concentrations were more effective than those that absorbed higher ionic concentrations.

PART E -- POST-LAB QUESTIONS

1. Define enzyme. What group of macromolecules do enzymes fall under?
An enzyme is a protein that acts as a catalyst, which speeds up the rate of chemical reactions.

2. What is the name of the enzyme used in this lab? What was the substrate?
The name of the enzyme used in this lab is called catalase and the substrate was hydrogen peroxide.

3. Write the reaction for this lab; Highlight reactants in YELLOW and products in LIGHT BLUE.
2H2O2 ----> 2H2O + O2

4. What does it mean to “denature a protein”? What factors can cause proteins to denature?
Denaturing a protein means modifying the molecular structure of a protein. Chemical reactions can cause
proteins to denature.

5. A similar experiment is performed similar to this lab. The person uses 3 pieces of liver that weigh
0.5 gram, 1.0 gram and 2.0 grams. In terms of reaction rate, predict what you feel the results of
this experiment would be. Explain your reasoning.
The chemical reactions with heavier liver pieces would happen faster and end faster because there are
more enzymes available to speed up the reaction and vice versa with the lighter liver pieces.