Kendriya Vidyalaya

Session-2016-2017
Study Of digestion Of Starch By
Salivary Amylase & Effect Of
pH & Temperature On It
Summited To
Summited By(P.G.T- Chemistry)
Class -12
Roll No.-

Certificate
This is to certify that is a bonafide
student of class XIIth Science during
session 2016-2017. He has
completed his project on Study of
digestion of starch by salivary
amylase and effect of pH and
temperature on it successfully under
my guidance and supervision.

(Principal)
(P.G.T-Chemistry)

Acknowledgement
Acknowledgement is the most beautiful page in any projects
final pages. More than a formality, this appears to me the best
opportunity to express my gratitude.
I must say thanks to my parents who provide me sufficient
money and help in making of this project.
Mr. Sangram Banerjee, you are real foundation of the project
done by me. My project would never have been possible without
your guidance. Thank you very much.
Special thanks to my principal who gave support to me and
library which provides me helpful books related to this project.
Next on my thanks lists are my friends, teachers, students
who really showered constructive feedbacks and suggestions
without which this project would not have been in present form.

INDEX

S.
No.

Contents

Objective

Introduction

Apparatus

Materials and Chemicals

Theory

6
Effect of Different Temperatures on the Activity of Salivary Amylase on
Starch
7
Effect Of Different pH On The Activity of Salivary Amylase On Starch
8

Bibliography

Objective
Our aim is to study the effect of different temperatures and pH on the activity of
salivary content, amylase on starch.

Temperatur
e

Conditio
n

pH

Introduction

An amylase is
an enzyme that catalyses the hydrolysis of starch into sugars.
Amylase is present in the saliva of humans and some other
mammals, where it begins the chemical process of digestion.
Foods that contain large amounts of starch but little sugar,
such as rice and potatoes, may acquire a slightly sweet taste
as they are chewed because amylase degrades some of their
starch into sugar. The pancreas and salivary gland make
amylase (alpha amylase) to hydrolyze dietary starch
into disaccharides andtrisaccharides which are converted by
other enzymes to glucose to supply the body with energy.
Plants and some bacteria also produce amylase. As diastase,
amylase was the first enzyme to be discovered and isolated
(by Anselme Payen in 1833). Specific amylase proteins are
designated by different Greek letters. All amylases
are glycoside hydrolases and act on -1, 4-glycosidic bonds.

Classification

-Amylase
The -amylases (EC 3.2.1.1 ) (CAS# 9014-71-5) (alternative names: 1,4-D-glucan glucanohydrolase; glycogenase) are calciummetalloenzymes,
completely unable to function in the absence of calcium. By acting at
random locations along the starch chain, -amylase breaks down longchain carbohydrates, ultimately
yielding maltotriose and maltose from amylose, or
maltose, glucose and"limit dextrin" from amylopectin. Because it can
act anywhere on the substrate, -amylase tends to be faster-acting
than -amylase. In animals, it is a major digestive enzyme, and its
optimum pH is 6.77.0.
In human physiology, both the salivary and pancreatic amylases are amylases.
The -amylases form is also found in plants, fungi (ascomycetes and
basidiomycetes) and bacteria (Bacillus).

-Amylase
Another form of amylase, -amylase (EC 3.2.1.2 ) (alternative names: 1,4--Dglucan maltohydrolase; glycogenase; saccharogen amylase) is also synthesized
by bacteria, fungi, and plants. Working from the non-reducing end, -amylase
catalyzes the hydrolysis of the second -1,4 glycosidic bond, cleaving off two
glucose units (maltose) at a time. During the ripening of fruit, -amylase breaks
starch into maltose, resulting in the sweet flavor of ripe fruit.
Both -amylase and -amylase are present in seeds; -amylase is present in an
inactive form prior to germination, whereas -amylase and proteases appear once
germination has begun. Many microbes also produce amylase to degrade
extracellular starches. Animal tissues do not contain -amylase, although it may be
present in microorganisms contained within the digestive tract. The optimum pH
for -amylase is 4.05.0[5]

-Amylase
-Amylase (EC 3.2.1.3 ) (alternative names: Glucan 1,4--glucosidase;
amyloglucosidase; Exo-1,4--glucosidase; glucoamylase; lysosomal -glucosidase;
1,4--D-glucan glucohydrolase) will cleave (16) glycosidic linkages, as well as the
last (14)glycosidic linkages at the nonreducing end of amylose and amylopectin,
yielding glucose. The -amylase has most acidic optimum pH of all amylases because
it is most active around pH 3.

Apparatus

Ice
Cubes
Bunse
n
Burne
r

Therm
omete
r

pH
Tablet
s
Test
Tube
Water

Dropp
er

Material & Chemicals

Required

NaCl Solution

Iodine Solution

Saliva Solution

Starch Solution

Theory

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 by the process of
digestion that breaks down the complex substance of starch into simpler molecules
of glucose, which are further metabolized into CO2 and water through the process
of glycolysis. The human digestive tract starts at the mouth and ends at the anus.

In the Beginning
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 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 activity of enzymes is strongly affected by several factors, such as

temperature and pH

Effect of Temperature

All enzymes are proteinaceous in nature. At a lower temperature, the enzyme

salivary amylase is deactivated and at the higher temperature, the enzyme is
denaturated. Therefore, more time will be taken by an enzyme to digest the starch
at lower and higher temperatures. Optimum temperature for the enzymatic
activity of salivary amylase ranges from 32 C to 37 C. The optimum temperature
means that the temperature at which the enzyme shows the maximum activity. At
this optimum temperature, the enzyme is most active and hence, takes less time to
digest the starch.

Effect of pH
The optimum pH for the enzymatic activity of salivary amylase ranges from 6 to 7.
Above and below this range, the reaction rate reduces as enzymes get
denaturated. The enzyme salivary amylase is most active at pH 6.8. Our stomach
has 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 Different Temperatures on the

Activity of Salivary Amylase on Starch
Procedure

Take beaker containing 15 ml of 1% starch solution + 3 ml of 1% NaCl solution.

Divide and pour this solution into three test tubes and mark them as A, B and C.

Maintain the temperature of the beaker containing ice cubes at 5C.

Take beaker containing ice cubes and keep it on the table.

Take another two beakers containing water and heat over the Bunsen burner.

Now transfer experimental tube A into a beaker containing ice.

Transfer the second experimental tube B into water bath set at 37C and third
experimental tube C into the beaker maintained at 50C.

Using a dropper, take 1 ml saliva solution and transfer the solution into test tube A.

Similarly, add 1 ml saliva solution into test tube B and test tube C.

Immediately, using a dropper, take few drops from experimental tube A and transfer this
into first series of test tubes having iodine solution.

Similarly, using fresh droppers, do the same procedure for test tube B and test tube C and
transfer the solution into second and third series of test tubes having iodine solution.

Note this time as zero minute reading.

After an interval of 2 minutes, again take a few drops from each tube and add to the iodine
tubes and note the change in colour of iodine.

Keep on repeating the experiment at an interval of every 2 minutes till colour of iodine does
not change.

Results
It takes less time to reach achromic point at 37C, as the enzyme is maximum active at this
temperature, while at higher and lower temperatures more time is taken to reach the achromic
point.

Conclusion
All enzymes are proteinaceous in nature. At lower temperatures, the enzyme salivary amylase is
deactivated and at higher temperatures, the enzyme is denaturated. Therefore, more time will be
taken by enzyme to digest the starch at lower and higher temperatures. At 37 C, the enzyme is
most active, hence, takes less time to digest the starch.

Effect of Different pH on the Activity of

Salivary
Amylase on Starch
Procedure

Take a beaker containing 15 ml of 1% starch solution + 3 ml of 1% NaCl solution.

Divide and pour this solution into three test tubes and mark them as A, B and C.

Add pH tablet 5 into test tube A, pH tablet 6.8 into test tube B and pH tablet 8 into test
tube C.

Now transfer experimental tube A, B and C into a beaker containing water and a
thermometer for recording temperature. Temperature of this beaker is to be maintained at
37C.

Using a dropper, take 3 ml saliva solution and add 1 ml of solution to each of the three test
tubes.

Immediately using a dropper, take few drops from experimental tube A and transfer this
into the first series of test tubes having iodine solution.

Similarly, do the same procedure for test tube B and test tube C and transfer the solution
into second and third series of test tubes having iodine solution.

Note this time as zero minute reading.

After an interval of 2 minutes, again take a drop from each tube and add to the iodine tubes
and note the change in colour of iodine.

Keep on repeating the experiment at an interval of every 2 minutes till colour of iodine does
not change.

Results
pH 5 is acidic and pH 8 is alkaline, therefore salivary amylase did not act in these tubes. Whereas,
the enzyme acted in the tube with pH 6.8 (i.e., slightly acidic) and digested the starch.

Bibliography
Books:
Laboratory Manual in Biology by Dr. J.P Sharma (Plus Two): - Laxmi Publications (P) Ltd.

Websites:

http://www.britannica.com/science/amylase

http://www.eng.umd.edu/~nsw/ench485/lab5.htm

https://en.wikipedia.org/wiki/Amylase

http://www.pua.edu.eg/PUASite/uploads/file/Pharmacy/Courses/fall%202013/PHR
%20374/practical%2012.pdf

https://www.apsu.edu/sites/apsu.edu/files/chemistry/SP11_1021_BREAKING_DOWN_STA
RCH_USING_SALIVARY_ENZYMES.pdf