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Aim

Studying the effects of


temperature on the
activity of salivary
amylase
Problem
statement
What are the effects of
temperature on the
activity of salivary
amylase on starch ?
Variable
Variable
 Manipulated variable : temperature
of the medium
 Responding variable : rate of
enzymatic reaction
 Constant variable :volume of saliva
(enzyme)
Hypothesis
The rate of the activity
of salivary amylase on
starch increase with the
increase in temperature
until it reaches the
optimum temperature
of 37 C
Materials
1% starch
suspension , saliva
solution , iodine
solution , ice and
distilled water
Apparatus
Beakers , test tube , syringes
,dropper , glass rods , white tiles
with grooves ,thermometers ,
Bunsen burner , tripod stand , test
–tube rack ,wire gauze and
stopwatch
Technique
Test for the presence of starch
using the iodine test ,measure
and record the time taken for
the hydrolysis of starch to be
complete with a stopwatch .
Procedure
1 )The salivary collected in a beaker is diluted with an
equal volume of distilled water .(Rinse your mouth
before collecting the saliva )
2 ) 5ml of 1% starch suspension is added to each test
tube , labelled A(1) , B(1) ,C(1) ,D(1), and E(1)
respectively with the aid of a syringe .
3 ) 2 ml of saliva solution each is add to another set of
test tube , labelled A(2) , B(2) , C(2) , D(2) , and E(2)
respectively , using a new syringe .
4 ) Test tube A(1) and A(2) , B(1) and B(2) ,C(1) and
C(2) , D(1) and D(2) , E(1) and E(2) are immersed
respectively into five different water baths with
temperature which are kept constant at 0 C , 28 C , 37
C , 45 C and 60 C .
5 ) The test tube are left for 5 minutes .
6 ) A drop of dilute iodine solution is placed into
each groove of the white tile .
7 ) After 5 minutes , the starch suspension in test
tube A(1) is poured into the saliva solution in
test tube A(2) .
8 ) The mixture is stirred using a glass rod . A
stopwatch is activated and the time is recorded
as zero minutes .
9 ) A drop of mixture from test tube A(2) is
immediately placed in the first groove of the tile
containing iodine solution .
10 ) The iodine test is repeated every minute for
10 minutes .The dropper is rinsed in a beaker of
distilled water after each sampling .
11) The time taken for the hydrolysis of starch
to be completed is recorded , that is , when the
mixture no longer turns blue-black in colour
when tested with the iodine solution .
12) The test tube containing the mixture are kept
in their respective water bath through the
experiment .
13 ) Step 7 to 10 are repeated for test tube B , C ,
D , and E
14 .Thermometers are used to
ensure that the temperature remain
constant throughout the experiment .

15 ) The results are recorded in the


following table and a graph showing
the rate of enzymatic reaction , 1/t
against temperature (C) is plotted .
1/t against temperature
Test Temperatur Time taken for Rate of
tube e the hydrolysis enzymatic
(C) of starch to be reaction 1/t
completed (min-1)
(minutes
A(2) 0 No complete 0
after 10 minutes

B(2) 28 2 0.50
C(2) 37 1 1.00
D(2) 45 3 0.33
E(2) 60 No complete 0
after 10 minutes
Discussion
1) The test tube are immersed in their respective
water bath for 5 minutes at the beginning of the
experiment to allow both solution to reach the
temperature set .
2) Starch is hydrolysed by salivary amylase to a
reducing sugar (maltose) .
3) The Iodine solution is used to test for the
presence of starch . If the iodine solution
changes from brownish-yellow to blue black ,
this indicates that all the starch has been
hydrolysed by the salivary amylase .
4) The graph shows that the rate of enzyme activity
against temperature is a bell-shape curve .
a) At low temperature , for every 10 C increase in
temperature , the rate of enzymatic reaction is
doubled .
b) the maximum rate of reaction occurs at 37 C
,which is the optimum temperature for salivary
amylase .
c) The average human body temperature is also 37
C.
5) At 0 C, the enzymatic is not active. Therefore,
the salivary amylase cannot hydrolyse the starch .
6) As the temperature increase , the rate of
enzymatic reaction increase until it reach an
optimal temperature of 37 C .
7) The optimum temperature for the activity of
salivary amylase is 37C . This is because the
hydrolysis of starch is completed in a shorted
period .
8) Beyond the optimum temperature ,the rate of
enzymatic reaction decrease and ceases
altogether at 60 C . There is no enzyme activity
at 60 C . This is because the enzyme has
denatured due to the high temperature .
Conclusion
Change in temperature effect the
activity of salivary amylase on
starch .Salivary amylase in inactive
at 0 C and denatures at 60 C .The
rate of reaction catalysed by
salivary amylase is highest at 37 C
,which is the optimum temperature
.The hypothesisis accepted.