Hydrogen Peroxide Vs.

Catalse
Introduction:
The lab we conducted was to identify how long it takes for the enzyme catalase to digest
H2O2 into H2O and O2. The enzyme Catalase is the only enzyme that can do this because it has
the specific shape that corresponds to the shape of the substrate. Although it can digest on its
own, Catalase speeds up the process. We decided to test how pH would affect the change the rate
of the digestion. We decided to change the pH because enzymes work best in a specific range so
by altering its pH, we would change the rate at which how fast it can digest the substrate. My
group and I hypothesised that if the pH balance of the enzyme is decreased, then the enzyme will
not work as fast because the shape will not match the shape of the H2O2.

Catalase Lab Procedure

1. _____all_____Check that you have all the equipment below.
Computer with Internet access and Vernier LoggerPro software
LabQuest Mini
Vernier Gas Pressure Sensor
Two-hole black rubber stopper (top)
Plastic tubing with Luer-lock connectors
20-200 L micropippetor set to 100 L
micropipette tips
50mL graduated cylinder
125 mL or 250 mL Erlenmeyer flask
Magnetic stirrer
Stirring bar

Thermometer
Metal clamp stand
At the central table:
Hydrogen peroxide
Catalase enzyme suspension
2. ____all______Check that every person is wearing apron and goggles.
3. ____all______Check that everyone who will touch the enzyme or liquids is
wearing thick latex gloves (blue and yellow gloves)
On the blank lines, write the initials of the teammate who completed the task:
4. __________Open your laptop and open Logger Pro .
5. __________Click File Open and open the folder in Biology by
Vernier. Then choose 06 Enzyme (Pressure).
6. __________Connect the LabQuest Mini to the computer using the USB
cable, and connect the Gas Pressure Sensor to CH 1 of the LabQuest Mini.
Set up the laboratory apparatus as seen in the picture:
7. __________Measure out 50mL of 1.5% H2O2
and pour into an Erlenmeyer flask.
8. __________Carefully place a magnetic stir bar in
the flask.

9. __________Place the flask on a magnetic stir

plate. Use a clamp to fasten the flask to the ring
stand as shown. Position the flask at the center
of the magnetic stirrer.
10. __________Test the stirrer at 100 rpm.
11. __________Stop the stirrer.
12. __________Use the plastic tubing with two Luerlock connectors to connect the two-hole rubber
stopper assembly to the Gas Pressure Sensor as shown in the image. The
valve connected to the stopper should stay closed during this investigation.
Its closed when its flat, parallel to the floor
Complete all the steps below quickly to complete your test reaction.
13. __________Start data collection: click green Collect button on Logger Pro.
14. __________Using a micropipette, add 100 L of enzyme suspension to the
contents of the flask.
15. __________IMMEDIATELY tightly seal the flask by placing the stopper in and
HOLDING it in carefully.
16. __________IMMEDIATELY Turn the stir plate on to 100.
NOTE: If the pressure exceeds 130 kPa, the pressure inside the flask will be
too great and the rubber stopper will likely pop off. HOLD DOWN the

stopper or be ready to have uncovered chemicals on your desk

WAIT 200 seconds (3.3 minutes) while data is
collecting- Do NOT click STOP. Data collection will automatically stop
after 200 seconds.
17. __________Turn off the stir plate.
18. __________Carefully remove the stopper from the flask to relieve the
pressure.
19. __________Use a thermometer to test and record the temperature of the
liquid in your lab packet
20. __________Pour all chemical waste into a RED bucket.
21. __________Remove the magnetic bar.
22. __________ Dispose of your used micropipette tip.
23. ___all_______Take off your safety gear and place it neatly away.
24. Observe the graph generated using LoggerPro software (example shown
below).
25. Hold down Control on the keyboard and press j to zoom in the graph.
26. Highlight the section of the graph where the slope is increasing, by clicking
and dragging your mouse across it.
27. Click the Analyze tab at the top of the page, and choose Linear Fit. A

statistics box will appear for your highlighted section of the graph.
28. Record the slope of the line, m, as the rate of catalase activity in kPa/s in
your lab packet (page 13)
29. Click File, Save As, and save with the file name:
(Your Names) Sam, Anabel, Jenni, Jose Control Trial Logger Pro Data
Email the graph to Ms. Lenowitz by following these final steps!
30. Click File and choose Print Graph. In the drop down menu of printers, choose
Cute PDF Writer in the drop down list. This will export your graph as a
PDF file. Be patient- it takes a few seconds to pop up.
31. Save the PDF with the this file name:
(Your Names) Sam, Anabel, Jenni, Jose Control Trial Data PDF
32. Open your MESA email and Compose a new email to
glenowitz@mesacharter.org
33. Attach the PDF to the email.
34. Press send!
Data Analysis:
The overall results support our hypothesis because our slope decreased after the
experiment trial. The slope decreased from 581.9 (control trial) to 579.1 (experimental trial).
When the shape of the enzyme changed because of the pH, it took longer to bind with the H2O2
which decreased the reaction rate.

Conclusion:
One problem that occurred in the lab procedure is that the first time we tried to collect the
data but the gas pressure sensor wasnt working properly so we could not collect any data for the
control. Our hypothesis was supported because we successfully decreased the reaction time. Next
time, to improve the experiment, I would lower the pH level to 1.