BIOCHEMISTRY LABORATORY

Name: MAANO, KRISLYN DIANE P. Date: __________________

Group: __________________________________ Rating:_________________

Laboratory Activity 9
FACTORS INFLUENCING ENZYME ACTION

I. DISCUSSION:
Enzymatic Activity
Optimum Conditions
Activators and Inhibitors

II. PROCEDURE:

A. SUBSTRATE CONCENTRATION
1. Label 8 test tubes 1, 2,3,4,5,6,7 and 8 and proceed.
2. Add 1 ml. of enzyme extract to each tube.
3. Add the substrate hydrogen peroxide(H2O2) as follows
Test tube 1= 0 drop
Test tube 2=3 drops
Test tube 3=6 drops
Test tube 4=9 drops
Test tube 5=12 drops
Test tube 6=15 drops
Test tube 7=18 drops
Test tube 8=21 drops
4. Examine the bubbles and the extent it travels on the tube.

Result: In terms of the height of the foam after 60 seconds.

Test tube 1= 0 drop
Test tube 2=3 drops
Test tube 3=6 drops
Test tube 4=9 drops
Test tube 5=12 drops
Test tube 6=15 drops
Test tube 7=18 drops
Test tube 8=21 drops

Conclusion: Therefore, the experiment suggests that the more drops of substrate
hydrogen peroxide substance to the enzyme, the higher the foams reach in the test
tubes.

B. ENZYME CONCENTRATION
1. Label 3 test tubes 1, 2 and 3 proceed as follows:
TEST TUBE 1= Place 1 ml enzyme
TEST TUBE 2= Place 2 ml enzyme
TEST TUBE 3= Place 3 ml enzyme
2. Add the same amount of substrate hydrogen peroxide in each test tube and observe. 3.
Examine tubes.

Result: Once hydrogen peroxide was added with the same amount in each test tubes
from started to be evident due to reaction of the chemical after 20 seconds that is
because of the work of the catalyst which has the presence of oxygen.
 Conclusion:

Therefore, when hydrogen peroxide is added to enzyme mixture there is a reaction of

catalyst that has the presence of oxygen and every mixture.

C. TEMPERATURE
1. A measured amount of starch solution is placed into 3 test tubes.
2. Label 3 test tubes A, B, and C that are immersed in water and kept at different temperatures.
Temperatures:
Tube A: 5 deg C
Tube B: 20 deg C
Tube C: 35 deg C

3. The same amount of salivary amylase is placed into these test tubes.
4. The spotting tile with iodine solution will monitor the reaction.
5. After 1 minute, a drop of solution will be put in the tile. Repeat every minute.

Result:
TIME TEST TUBE 1 TEST TUBE 2 TEST TUBE 3

1 MINUTE BLACK BLACK BLACK

COLORATION COLORATION COLORATION

2 MINUTES BLACK STARCH TURNED STARCH TURNED

COLORATION TO SUGAR TO SUGAR
(TRANSPARENT) (TRANSPARENT)

3 MINUTES BLACK STARCH TURNED STARCH TURNED

COLORATION TO SUGAR TO SUGAR
(TRANSPARENT) (TRANSPARENT)

4 MINUTES BLACK STARCH TURNED STARCH TURNED

COLORATION TO SUGAR TO SUGAR
(TRANSPARENT) (TRANSPARENT)

Conclusion: Therefore, the higher the temperature gets the more efficient enzymes turns into starch
into sugar
 D. pH
6. Prepare 6 clean test tubes and label 1, 2, 3, 4, 5,
6,.Label test tubes 1=ph7, 2=ph 9 and
3=ph11. Place 2ml amylase in the labelled
tubes.
7. Place 1 ml starch in the other 3 tubes. Add 1 drop of
iodine to each starch tube.
8. Pour 1 tube of starch solution into each amylase tube.
9. Start timer
10. Record observation at each time interval.
TEST 10 seconds 30 seconds 2 minutes 3 minutes 4 minutes 5 minutes
TUBES

1 with ph 7 BLACK BLACK BLACK BLACK BLACK BLACK

COLORATIO COLORATIO COLOR COLOR COLOR COLOR
N N

2 with ph 9 BLACK BLACK BLACK STARTED TO MIXTURE TRANSPARE

COLORATIO COLORATIO COLORATIO LOSE BECAME NT
N N N COLOR TRANSPARE
NT

3 with ph 11 BLACK STARTED TO MIXTURE MIXTURE MIXTURE TRANSPARE

COLORATIO LOSE BECAME BECAME BECAME NT
N COLOR TRANSPARE TRANSPARE TRANSPARE
NT NT NT

Result: it is a period of time the mistutes because as a result of the chemical change in the mixture.

Conclusion: therefore, the higher the pH level of the enzymes the faster it is because transparent as an
evidence of a positive result of the experiment.

III. QUESTIONS:
1. What are the optimum conditions for enzyme activity in the human body?
There is a certain temperature at which an enzyme's catalytic activity is at its
greatest (see graph). This optimal temperature is usually around human body
temperature (37.5 oC) for the enzymes in human cells.

2. Give other factors which influence enzyme activity and state the effect of each factor.
Enzyme activity can be affected by a variety of factors, such as temperature, pH,
and concentration. Enzymes work best within specific temperature and pH
ranges, and sub-optimal conditions can cause an enzyme to lose its ability to
bind to a substrate.