CONCLUSION

The process of digestion of starch by salivary amylase is well

studied as the effect of temperature. pH and disinfectant on
the activity of salivary amylase on starch was well
understood. The optimum temperature for the enzymatic
activity of salivary amylase is 37 C, and the optimum pH for
the enzymatic activity of salivary amylase is 7 while there
was no detectable disappearance of starch using disinfectant
above and below this range, the reaction rate reduces as
enzymes get denaturated. the above analyses is proven to be
right due to the color changes at different temperatures and
pH, meanwhile 37C temperature, and the known reference
for amylase and iodine test.
INTRODUCTION
Enzymes are synthesized in a living cell and speed up
reactions efficiently decreasing the energy of activation. The
effectiveness of enzymes as catalysts is demonstrated by the
very high reaction rates at physiological temperatures. In the
following experiments, the highly specific nature of this is
shown. Enzymes are extracted from plants and biological
materials by different methods. Amylase is an enzyme found
in human saliva and pancreas. It is the digestive enzyme that
is needed to break down starch molecules. Amylase must be
kept at a certain condition to function at its optimum level.
This experiment will explore the effect of PH, temperature,
and disinfectant on the activity of amylase to hydrolyze the
starch.
Starch is mainly composed of two components, and
amylopectin presents a great challenge to hydrolytic enzyme
systems. This is due to the residues involved in a- 1,6-
glycosidic links yet the a-1,6- glucosidic links must also be
cleaved for the complete hydrolysis of amylopectin to
glucose.

MATERIALS
Equipment

1)Triple beam balance

 2) spot plate
3) 100- ml beaker
4) hot plate
5) thermometer
6) 10ml test tubes

REAGENT
 Lysol
 NaHCO3
 2% starch
 PH 4 buffer solution
 PH 10 buffer solution
 PH 7 buffer solution
 25-ml graduate cylinder
 Amylase
 0.2 M citric acid
 0.2 KH2PO4
 0.01 M NaOH
 0.5% NaCl
 Iodine solution
 Starch solution
PROCEDURE

 Preparation of a-amylase from saliva

In a 25-graduated cylinder, 5 ml of saliva Added 15
ml of distilled water, and 5 ml of 0.5% NaCl were
also added.
 Preparation of starch solution
 10g of soluble corn starch was mixed in 50 ml of
cold water.
 While stirring the slurry was added to 400 ml of
gently boiling water in a large beaker.
 The gelatinized starch solution was well mixed
and cooled to room temperature.
 More water was added to bring the total volume
to 500 ml
 More water was added to bring the total vol to
500 ml
 A few drops of the starch solution were placed on
a glass plate. 1 drop of iodine reagent was added
and a deep was observed developing.

 Preparation of buffers
 PH 4: 0.2 M citric acid was adjusted acid to PH
14 with NaOH.
 PH 7:0.2 M monosodium dihydrogen phosphate
was adjusted to PH 7 with NaOH.
  PH 10: 0.2 M sodium bicarbonate was adjusted
to PH 10 with NAOH.
 Effect of PH
 Three test tubes were prepared and labeled 1-3.
To the first test tube, 1 ml of buffer (pH 4) was
added; to the second, pH 7 buffer, and to the
third, pH 10 buffer. To each tube 1ml of 2%
starch was added
 Another three test tubes were prepared and 2
ml. of enzyme solution was placed in each test
Etube. All six tubes were incubated at 37 C for
10 minutes.
 Pairs of tubes were mixed and the reaction at
once was monitored at the one-minute interval
for 10 min by taking 3 drops and it was
transferred to a spot plate containing iodine
solution. The change in color of the iodine was
observed.

 Effect of temperature
2 ml of enzyme solution was placed in three
separate test tubes and 2 ml of buffered 1% starch
(pH7) solution in another 3 separate test tubes.
Pairs of tubes were incubated for 10 min at either
0,37 or 70, then mixed and processed as in
procedure D (3). The change in color in the iodine
solution was observed.
  Effect of disinfectant.
In one test tube, one drop of Lysol disinfectant was
Added and 2ml of enzyme solution and at 37C it
was pre-incubated for 10min along with 2 ml of 1%
of buffered starch in a separate tube. the two test
tubes were mixed immediately and processed as
in procedure D (3). The change in color in the iodine
solution was observed.

RESULT

o Effect of pH
P
H
1 2 3 4 5 6 7 8 9 10

4 bro Bro bro bro Dar Dar Dar Dar blac Blac
wn wn wn wn k k k k k k
bro bro bro bro
wn wn wn wn
7 Sligh Slih Ligh Ligh blu blu Dee Dee bro Bro
 tly tly t t e e p p wn wn
blue blu blu blu blu blu
e e e e e
Blac Blac Bla Bla blac blac blac blac blac Blac
k k ck ck k k k k k k

Ph 4 ph7 Ph 10

A brown color was recorded for the first time in 4 minutes

dark brown for the 5-7th minutes, and dark color for 9 and
10th minutes for the pH 4 test. For pH 7 and 8th minutes,
brownish blue for 9th and brown for 10th minutes respectively.
While pH 10 showed a black color all through.

o Effect of temperature
Tempr
ature
 1 2 3 4 5 6 7 8 9 10
0 bro bro bro bro Dar Dar Dar Dar bla bla
wn wn wn wn k k k k ck ck
bro bro bro bro
wn wn wn wn
37 Slig Slig Lig Lig blu Blu De De bro bro
htly htly ht ht e e ep ep wn wn
blu blu blu blu blu blu
e e e e e e
70 Bla Bla bla Bla Bla Bla Bla Bla Bla bla
ck ck ck ck ck ck ck ck ck ck

for the effect of temperature 0 record brown for the first 4

minutes, dark brown for 5-8th minutes, and black for 9 and
10th minutes. For 37C slightly blue color for the first 2
minutes, light blue for 3 and 4th minutes, blue color for 5 and
6th minutes, and deep blue for 7 and 8th minutes, brownish
blue for 9th and brown for 10th minutes respectively. While
70C also showed a black color all through.

Discussion

All living beings need energy to survive. It is from the food we

consume that we get our energy. We know that the energy
we are getting is from the process of digestion that breaks
down the complex substances of starch into simpler
molecules of glucose, which are further metabolism into CO2
and water through the process of glycolysis. The human
digestive tract starts at the mouth and ends at the anus. The
digestion of the food starts as soon as we put food in our
mouth. Our teeth cut the food into small pieces and the
salivary glands secrete saliva glands secrete saliva that mixes
with these food materials. The saliva contains an enzyme
called salivary amylase. Which hydrolyses starch into
maltose. The complete digestion of starch occurs only in the
small intestine by the action of pancreatic amylase.
The effect of temperature, pH, and disinfectant on the
activity of salivary amylase.

Effect of ph
The above result obtained on the effect of pH on the
enzymatic activity of salivary amylase, we observed that
the optimum pH on the enzymatic activity of salivary
amylase is pH 7 this is because there is theoretical proof
that the positive test to that above and below this
range, that pH 10 and pH 4 the reaction rate reduces as
enzymes get denatured. The enzyme salivary amylase is
most active at pH 6.8. our stomach has a high level of
acidity which causes the salivary amylase to denature
and change its shape. so the salivary amylase does not
function once it enters the stomach.
Effect of temperature
All enzymes are proteinaceous in nature. At a lower
temperature, the enzyme salivary amylase to
deactivated, and its a higher temperature, the enzyme is
denatured. Therefore, more time will be taken by an
enzyme to digest the starch at lower and higher
temperatures. From the results obtained on the effect of
temperature from above result obtained on the effect of
temperature on the enzymatic activity of salivary
amylase, we observed that the optimum temperature
for the enzymatic activity of salivary amylase is 37C. This
is also because there is theoretical proof that the
positive test of salivary amylase on iodine is blue and
only at 37C temperature showed a positive test to that
optimum temperature for the enzymatic activity of
salivary amylase ranges from 32C to 37C. To optimum
temperature means that the tempreture at which the
enz