UP-FA2-BSN1-16

BATAC, JOSHUA KYLE C.

PHINMA UNIVERSITY OF PANGASINAN

COLLEGE OF ALLIED HEALTH SCIENCES
DEPARTMENT OF NURSING

ACTIVITY 8 - 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:
Test tube
1-Only the solution's surface had a tiny coating of bubbles.
2-It has a coating of bubbles almost an inch thick, almost like white foam.
3-It features a bubble or white foam layer that is slightly thicker than an inch.
4-It has a bubble or white foam layer that is around 1.5 inches thick.
5-It has a thick layer of white froth or bubbles that is around 6.5 inches thick.
6-It has a roughly 7.5-inch-thick coating of white froth or bubbles on top.
7-A bubble or white foam layer covering it has a thickness of roughly 6.5 inches.
8-It shows a layer of white foam or bubbles that is roughly 6 inches thick.

Conclusion:.As substrate concentration increases, more substrate and enzyme collisions

occur, increasing the reaction rate and producing more products. However, once the
enzyme has been saturated with the substrate, further additions have no effect on the rate
because the catalytic site is occupied, and the enzyme's activity is the only factor
determining how much product is produced.
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:
Test tube 1:It produces the thinnest layer of white foam or bubbles among the results after 20
seconds.
Test tube 2:It creates a coating of bubbles or white foam that is nearly twice the thickness after 20
seconds.
the first tube's laver's thickness
Test tube 3:It produces a coating of bubbles or white foam after 20 seconds that is roughly three
thicker than the laver in the first tube by many times

Conclusion: There is a direct relationship between the two variables (enzyme

concentration and activity) as long as the enzyme concentration is less than the substrate
concentration. This indicates a proportionate relationship between the rate of the reaction
and the enzyme concentration. Simply put, adding an enzyme increases the rate by one
reaction per unit of time, while removing an enzyme decreases the rate by one reaction per
unit of time.

C.TEMPERATURE
Ameasured amount of starch solution is placed into 3 test tubes.
4. 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
5. The same amount of salivary amylase is placed into these test tubes.
6. The spotting tile with iodine solution will monitor the reaction.
7. 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 The iodine solution The iodine solution TIne Iodine solution
gets darker in color. gets darker in color. gets darker in color.
(Black (Black (Black)
2 MINUTES The iodine solution The iodine solution The iodine solution
gets darker in color. gets darker in color. gets a little darker In
(Black (Black color. (Dark brown)
3 MINUTES The iodine solution The iodine solution The iodine solution
gets darker in color. gets a little darker In gets very little darker
(Black color. (Dark brown) in color. (A little
lighter than dark
brown)
4 MINUTES The iodine solution The iodine solution The iodine solution
gets darker in color. gets very little darker gets less dark in color
(Black In color. (Lighter than than the previous one.
Dark brown) (A little lighter than
dark brown)

Conclusion:Heating starch to its boiling point causes it to degrade, resulting in shorter

dextrin chains and a color change. When glucose is reacted with iodine, no color change
occurs. The experiment reveals that amylase works best at 5 degrees Celsius but poorly at
higher temperatures such as 35 degrees Celsius. The iodine solution efficiently changes
color at 5 degrees Celsius, whereas at 20 and 35 degrees Celsius, two and three iodine
solutions, respectively, do not fully turn black.

C. 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 There’s no There’s no The top layer There is little There is little There is little
visible change visible change looks a little to no change in to no change in to no change in
white previous result previous result the previous
result.The
blackest
among the
others
2 with ph 9 The top and It closely The white The prior It continues to It’s getting
bottom are resembles the color at the white color grow whiter whiter, whiter
slowly turning prior outcome solution’s becomes even than test tube
white bottom is whiter one but not as
ascending white as test
upward tube three.
3 with ph 11 The top and The solution is Solution is The white Still getting It's getting
bottom now white than whiter than color is whiter whiter and has
displays a before.But before,yet becoming the whitest
more whitish some black some black opaque. color among
color than it’s color still color persists the others
test tube remains

Result: Each enzyme has a different pH range. When the pH of an enzyme is above or below its optimal,
its activity is reduced. The optimum pH of an enzyme is the same as the pH of the enzyme's natural
environment. For several enzymes, this corresponds to pH levels of around 7. However, according to the
results of the experiment, the highest pH level of 11 produced the fastest and most noticeable reaction at
all time intervals, while the lowest pH level of 7 produced no change at all.
 III. QUESTIONS:
1. What are the optimum conditions for enzyme activity in the human body?
-Enzymes typically function best in a specific temperature and pH range in the human body.
Close to body temperature (37°C or 98.6°F) and a neutral oH of 7 are ideal conditions for man-made
human enzymes. However. For maximum action, different enzymes may prefer different temperature
ranges and pH levels. Some enzymes are also dependent on specific cofactors or coenzymes. It is critical
that these ideal conditions remain as close to perfect as possible. because enzymatic denaturation or
inhibition is possible
2.Give other factors which influence enzyme activity and state the effect of each factor.
-Substrate Concentration: Increasing the substrate concentration often accelerates enzymatic
activity until all active sites are saturated. This assertion was debunked. More substrate has no
effect on reaction rate. As the concentration of the enzyme increases, more enzyme molecules are
available to catalyze the reaction, resulting in a faster reaction rate. provided there is sufficient
substrate.