Ayesha Nawab

STEM
Period 3
12/15/16
The Amazing Catalase Trials
The lab was conducted in order to separate H2O2 into H2O(water) and O2(oxygen).
Enzymes are proteins that increase the rate of reaction. Many enzymes are similar but have
specific substrates to bind with. The concept of this lab was to test how pH would change the
shape and affect the reaction. The enzyme's shape would deform and affect the rate of its
reaction. My group predicted that if the pH balance is decreased then the enzyme reaction rate
will not match the substrate and thus take long to react.

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 my groups hypothesis because our slope
decreased after the experimental trial. The slope went from 581
kPa/min(control) to 579 kPa/min(experimental). When the shape of the
enzyme changed due to the pH it toke it longer to find a substrate to bind
with and thus the reaction rate decreased.
Conclusion
One problem that occurred in the lab procedure is that the first time
we tried to collect the data but the the pressure sensor wasnt working
properly. We couldnt get our data for the control group. We could Improve
the experiment by having better materials to experiment on. Also, to have
more clear directions so we cant mess up the first time.