Enzymatic Activity of

Human Saliva against temperature

Dela Cruz, Juan [1] Dela Cruz, Juana [1] Dela Cruz, Juane [1] Christian Jayvon C. Laluna [2]

[1]
SHS student, Senior High School Department, Pasay City National Science High School.
[2]
Biology Instructor, Senior High School Department, Pasay City National Science High School.

ABSTRACT

INTRODUCTION activity decreasing at values above and

An enzyme is a protein molecule that below that point due to denaturation. For

is a biological catalyst with three enzymes, denaturation can be defined as the

characteristics. First, the basic function of an loss of enough structure rendering the

enzyme is to increase the rate of a reaction. enzyme inactive. This is not surprising

Second, most enzymes act specifically with considering the importance of tertiary

only one reactant, called a substrate, to structure in enzyme function and non-

produce products. The third and most covalent forces in determining the shape of

remarkable characteristic is that enzymes are enzymes [2].

regulated from a state of low activity to high Salivary amylase is the enzyme

activity and vice versa [1]

. The activity of produced by the salivary glands. Formerly

enzymes is strongly affected by changes in known as ptyalin, it breaks down starch into

pH and temperature. Each enzyme works maltose and isomaltose. Amylase, like other

best at a certain pH and temperature, its

enzymes, works as a catalyst. All catalysts enzyme solution was put in a large test tube

are enzymes, but not all enzymes are and labeled as 4°C. Two mL of the buffered

catalysts. A catalyst is a substance that starch solution (1% stach in phosphate

hastens a chemical reaction but does not buffer pH 6.7) was added in a separate large

become part of the product. Amylase digests test tube. Both the test tubes were incubated

starch by catalyzing hydrolysis, which is for ten minutes in an ice bath (4°C). The

splitting by the addition of a water molecule. solutions were immediately mixed. Three

The presence and absence of starch can be drops of the mixture was taken quickly and

confirmed by several tests such as the iodine two drops of the 0.001 M iodine solution

test, Benedict’s and Fehling’s test. In was added simultaneously onto a spot plate

general, a blue-black color indicates the (first well). This was the zero minute. After

presence of starch [3]. one minute interval (incubation continued),

The objectives of this experiment are three drops of the mixture was taken again

to examine the enzymatic activity and and two drops of the iodine solution was

specificity of salivary amylase depending on added simultaneously onto the second well.

changes temperature. This experiment also This was the one minute. Step 5 was

aims to determine the narrow range of repeated until a light-yellow colored

temperature and pH values at which salivary solution was observed. The time (t) was

amylase exhibits its optimum activity. noted. For the other temperatures (room

MATERIALS AND METHODOLOGY temperature, 37, 50 & 70 °C), steps 1 to 6

An enzyme solution was prepared by were repeated following the desired

mixing 1mL saliva with 9mL distilled water incubation temperature. The reciprocal of

and 30mL 0.5% NaCl. Two mL of the time (1/time, min-1) in step 6 versus the
temperature (T) was plotted. The optimum activity of salivary amylase is affected by

temperature of the amylase was determined. temperature.

Temperature Time (t)

RESULTS (T) °C min 1/t (min1-)

The effect of temperature on the 4 °C ∞ 0

enzymatic activity of salivary amylase was Room temp. 32

1.00 1.000
determined by measuring the rates of °C

reaction in varying temperatures and pH. 37 °C 11.0 0.091

The 0.5% NaCl added in the enzyme 50 °C 6.00 0.167

solution activates the salivary amylase to 70 °C ∞ 0

perform its function – to hydrolyze starch. Table 1. Results for the effect of

The hydrolysis or breakdown of starch due temperature on salivary amylase activity

to the action of salivary amylase is indicated Extreme temperatures cause the

by the change in color of the starch solution native folded structure of proteins to uncoil

from a blue-black color to a light-yellow into random configuration. As a result, the

colored solution. protein loses its biological enzymatic

DISCUSSION activity. This denaturization

Each enzyme has an optimum consequently, leads to loss of activity.

temperature at which it performs best. Figure 1 shows the graph of the reciprocal of

Below or above this temperature, the time against temperature based on the data

enzyme loses its functionality. Table 1 from Table 1.

shows the results obtained on how enzyme

 within these temperatures. After 37°C, the

graph then steeply declines as a result of loss

of activity. At 50°C and 70°C, salivary

amylase is denatured. The molecular

conformation of the enzyme becomes

altered as the hydrogen bonds responsible

for its secondary, tertiary, and quaternary

Figure 1. Plot of the reciprocal of time
structures are broken [4].
against temperature for the enzymatic

activity of salivary amylase

REFERENCES
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Role of Enzymes in Biochemical
The graph produced a bell-shaped
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[2]
Enzymes. (2011). Retrieved on January 3,
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http://users.rcn.com/jkimball.ma.ultranet/Bi
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[3]
Sethi, R. (2009). Biology. Rachna Sagar
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Pvt. Ltd.: New Delhi
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[4]
Solomon E., Berg, L. & Martin, D.
salivary amylase is suitable to function
(2005). Biology. Thomson Learning In