1.

Title

Effect of Temperature on Enzyme Activity

2. Introduction
A. purpose

This experiment investigates the effect of temperature on the structure of an enzyme, which
relates to the function and the amount of product formed. Enzymes are made of proteins that
are used as biological catalysts. Catalysts are substances that lower the amount of energy
necessary for a chemical reaction process. we observe how catechol oxidase will be tested at
various temperatures to determine the temperature range and the optimum temperature for
the formation of benzoquinone and how it denature and change the effectiveness of an
enzyme.

Catechol oxidase

Catechol + oxygen benzoquinone + H20

(Colorless substrates) + (enzyme in potato extract) (colored product)

B. Hypothesis

There will be more enzymatic activity when the temperature is at 40 0 C, than the reaction is at
the other temperatures. The formation of benzoquinone will be more evident in test tube 3C
tested in a warm bath at the presence of catechol oxidase.

3. Materials or procedure: steps

A. Materials

1. 5 test tubes
2. Test tube rack
3. Metric ruler
4. China marker
5. 1 % catechol
 6. Potato extract with catechol oxidase
7. Ice bath
8. Room temperature bath
9. 40°C water bath
10. 60°C water bath
11. Hot plate

B. steps (procedure)

1. Fill 400 ml beaker half full of water and add 3-4 boiling chips. Bring to boil on hotplate, then
turn down.
2. Label the top of five test tubes with "3A, 3B, 3C, 3D, 3E': and initials for identification.
3. Lay test tubes alongside a metric ruler and mark from the bottom at 1 cm and 2cm.
4. Fill each tube to the 1 cm mark with potato extract containing catechol oxidase, DO NOT MIX.
5. Place each tube in the respective water bath as listed below.
3A = ice bath
3B = room temperature
3C = 40·C water bath
3D = 60·C water bath
3E = 1 OO·C boiling water
Observe the thermometer in each water bath and record the actual temperature
6. Allow the test tubes to remain at the various temperatures for 5 minutes.
7. Remove the tubes after 5 minutes and add catechol to the 2-cm line of each tube. Swirl to
mix.
8. Immediately observe the color intensity (0-5) of each tube and record in the "0 min" column
of the table
9. Return the test tubes to the respective water baths for 10 minutes swirling occasionally to
mix.
10. Remove the tubes after 10 minutes.
11. Observe the color intensity (0-5) of each tube and record in the "1 0 min" column of the
table
12. Plot the data from the 10 minute on the graph in Figure
13. Discuss the results and record
14. Determine if the hypothesis was rejected or accepted

4. observations
A. Results

Fig 1. Color intensity at o minute

Fig 2. Color intensity at 10 minutes
 Temperature and Color Intensity

Actual Color Intensity Color Intensity

Tube Temperature °C O min. 10 min.

0-C ice bath Light yellow yellow

3A

20-C room Light yellow orange

3B temperature

40·C water bath yellow Orange brown

3C

60·C water bath Light yellow pinkish

3D

1OO·C boiling Cloudy white Cloudy white

3E water

Table 1: Effect of Temperature on Enzyme Activity

 Intensity Color intensity description

0 clear

1 Cloudy white

2 Light pink/yellow

3 pinkish

4 orange

5 Orange brown

Table 2: Color intensity scale

After adding potato extract catechol oxidase in each five test tubes and each was held at a
different temperature. The solution in test tube 3a turned light yellow at 0 min, and tuned
yellow at 10 minutes. The solution in test tube 3b turned light yellow at 0 min, and tuned
orange at 10 minutes. The solution in test tube 3c turned yellow at 0 min, and tuned orange
brown at 10 minutes. The solution in test tube 3d turned light yellow at 0 min, and tuned
pinkish at 10 minutes. The solution in test tube 3e turned cloudy white at 0 min, and
remained cloudy white at 10 minutes.
 6

3 0 Min
10 Min

0
0 20 40 60 100

Fig 3, Effect of Temperature(0,20,40,60,100) on Catechol Oxidase Activity versus color

intensity(0,1,2,3,4,5)

B. Conclusion

The benzoquinone was evident in test tube 3C shows the temperature for an enzyme to
function properly in a reaction. Deferent enzymes have their specific temperature they
function the most. So it can be concluded that the enzyme used in this experiment, catechol
oxidase, functions the most at 40 0 C of tube 3C.the hypothesis can be proving.

C. Error Suggestion for Improvement

Using test clamps at all times is advised when removing test tubes from boiling bath.

5. Works cited

-Lab manual (lab 7)

-Lab module and videos

- https://www.studocu.com/en-us/document/north-lake-college/biology-for-science-
majors-i/other/biology-lab-report-2/6164011/view
-https://www.chegg.com/homework-help/questions-and-answers/date-ch101-lab-
section-5d2-instructor-whil-30--effect-enzyme-concentration-starch-test-ime-
q20617187

https://www.google.com/search?
rlz=1C1CHBD_enET877ET877&sxsrf=ALeKk016ud0TQqdjqMEeV0lxiCpeI1eI5g:16061755
49505&q=catechol+oxidase+and+potato+extract+lab+report&sa=X&ved=2ahUKEwi3x8y
Y7pntAhVSz4UKHfjABWoQ1QIoAXoECBQQAg&biw=1517&bih=694&dpr=0.9
-https://quizlet.com/98085533/enzymes-lab-report-experimental-method-and-the-
action-of-catechol-oxidase-flash-cards/
-https://www.orusclothing.com/introducing-potato-enzyme-lab-report/