Enzymatic Activity of Salivary Amylase

Mark Mitchel N. Remorozo, Sabrina Ysabelle A. Reyes*, Chyna Mae P. Roca, Lemuel Salvador
Department of Psychology, University of Santo Tomas, Espana Boulevard, Manila

ABSTRACT: system of humans. Beta Amylase is used by

Salivary Amylase is an enzyme which is plants to process starch.
produced by the digestive process by
breaking down starch and converting it to
maltose, a smaller carbohydrate. In this
experiment, the effect of pH, that ranges at
4, 5, 6.7, 8 and 10, and temperature, that
ranges at 4 degrees, room temperature, 37
degrees, 50 degrees, 60 degrees and 70
degrees, to the enzymatic activity of
salivary amylase will be observed.

1. Introduction

Enzymes are to be found in any protein, Figure 1.​ Types of enzymes found in protein
most especially in globular proteins which
acts as a biological catalyst. A catalyst is a
substance that increases the rate of a 2. Materials and Methods
chemical reaction without itself undergoing
any permanent chemical change. The
2.1 Effect of Temperature
specific use of enzymes are for metabolic
reactions in the system.
A. Materials

The enzyme that was used in the Enzyme Solution (1mL saliva + 9mL distilled
experiment is known as ​“amylase” ​which H2O + 30mL 0.5% NaCl)
can be found in human or animal saliva.
This enzyme is used for the breaking down Buffered Starch (1% starch in phosphate
of starch which is essential for the digestive buffer pH 6.7)
process. There are two types of amylase
namely Alpha Amylase and Beta Amylase. 0.001 M Iodine Solution
Alpha Amylase is used for the digestive
Spot plate
 has been made and the optimim
Test tube temperature of the amylase was
determined.
Medicine Droppers

Constant Temerature bath (4 degrees,

Room temperature, 37 degrees, 50 degrees,
50 degrees and 70 degrees celsius)

B. Methods
Figure 2. ​ The procedure for temperature
In this experiment, the effect of
temperature and pH to the enzymatic
salivary amylase was obeserved. First, we
2.2 Effect of pH
will start with the effect of temperature.
Two large test tubes were used, labeled as
A. Materials
“A” and “B”. Test tube “A” contains 2mL of
the Buffered Starch Solution and test tube Enzyme Solution (1mL saliva + 9mL distilled
“B” contains the 2mL of Enzyme Solution. H2O + 30mL 0.5% NaCl)
Both were placed in a hot bath with the
assigned temperature for 10 minutes for 2% Unbuffered Solution
incubation. After incubating the 2 test
tubes, solution “A” and “B” were mixed as 0.001 M Iodine Solution
one. As the mixture was being created, two
drops of the Iodine Solution was placed in Acetate Buffer Solutions (pH 4 and 5)
each well of the spot plate. Three drops of
the mixture was then taken and placed in Phosphate Buffer Solutions (pH 6.7 and 8)
the first well on the post plate that contains
the iodine solution which was being Bicarbonate Buffer (pH 10)
observed as the zero minute. This was
repeated following the one-minute interval Spot plate
for each well until the solution is colorless
and until it has reached the 10 or 15 Test tube
minute. These steps were repeated for the
other temperatures (4 degrees, Room Medicine Droppers
temperature, 37 degrees, 50 degrees, 50
degrees and 70 degrees celsius). Finally, the Water bath set at 37 degrees celsius
plotting of the reciprocal time (1/t, min^-1)
B. Methods Table 1 presents the data collected in the
experiment for the effect of temperature to
The difference between the two mixtures is the enzymatic salivary amylase. The
that the test tube “A” contained 1mL of the enzymes work effectively at a certain
Unbuffered Starch + 1mL of the Acetate temperature wherein the reaction rate will
Buffer. The test tube “B” contained the be at a maximum.
same enzyme solution. The two test tubes Table 1. ​Experiment data in temperature
were incubated by hand for 10 minutes to TEMPERATURE RATE (1/t)
obtain the 37 degrees celsius. After
incubating the two test tubes, solution “A” 4 0
and “B” were mixed as one. As the mixture
was being created, two drops of the Iodine RT 0.13
Solution was placed in each well of the spot 37 0.20
plate. Three drops of the mixture was then
taken and placed in the first well on the 50 0.083
post plate that contains the iodine solution
which was being observed as the zero 60 0
minute. This was repeated following the
70 0
one-minute interval for each well until the
solution is colorless and until it has reached
the 10 or 15 minute. These steps were Table 2. ​Experiment data in pH
repeated for the other pH levels (4, 5, 6.7, 8 pH RATE (1/t)
& 10). Finally, the plotting of the reciprocal
time (1/t, min^-1) has been made and the 4 0
optimim pH of the amylase was
determined. 5 0.125
6.7 0.09
8 0
10 0.08

Figure 3. ​The procedure for pH

3. Results