Can Enzymes survive in the pH of 10?

Enzymes are proteins that speed up reactions like synthesis and digestion. Basically the
enzyme catalase lets the substrate (H2O2) so that it could be digested into H2O (Water) + O2
(Oxygen). Hydrogen peroxide is pretty toxic to to any living being organism, so we need to
digest it in order to prevent ourselves from getting extremely sick. Hydrogen peroxide is formed
whenever H2O and O2 is synthesized together with the catalase. When we digest, we break apart
the H2O2 and then separate the enzyme catalase from the H2O and the O2. In the lab, we had to
figure out the rate of reaction of the catalase as affected by the pH. So the prediction that my
team settled with was that if we increase the pH to 10, then the rate of reaction would increase.

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 uL 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. _____AG_____Open your laptop and open Logger Pro .

5. ____BG______Click File Open and open the folder in Biology by

Vernier. Then choose 06 Enzyme (Pressure).

6. ____MG____ Add 60 mL of pH 10.
7. __________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 Luer-

lock 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

Data Analysis
The results from our data did not support the original hypothesis if we increase the pH to
10, then the rate of reaction would increase. The slope of the data before we changed the pH was
0.008590/ kPa/min. But after we changed the pH to 10, the slope didnt change much. It went
down to -0.01136 kPa/min, which means that the pH of 10 killed the enzyme, and it had no
reaction. These results actually make sense because enzymes only works under a specific range
of the pH depending on the type of enzyme it is. This enzyme worked very well with the pH of
6.5. But the pH of 10 was too high and the function the enzyme had originally was changed
because of the pH.

Conclusion
There may have also been some contributing factors as to why we got the results we had
the first time. The first time, it couldve been because the reaction was delayed and the catalase
was entered in too early. The spinning rate also couldve been a factor because the speed
couldve messed up the reaction. The next time we do this experiment, we could possibly wait to
make sure everything is actually working correctly before even moving further in the process.