PANCREATIC

AMYLASE
Test 1

A.) IN THIS PROCEDURE, IDENTIFY THE

ENZYME, THE SUBSTRATE, AND THE
MANIPULATED VARIABLE.
Enzyme: AMYLASE

Substrate: STARCH

Manipulated Variable: TYPE OF ENZYME

B.) WHAT IS THE PRINCIPLE BEHIND THE

USE OF THE BENEDICTS AND BARFOEDS
TESTS?
Benedictss test- it is a test used to determine the presence of simple

sugar.

Barfoeds test- test used for the detection of monosaccharides.

REACTION INVOLVED

REACTION INVOLVED

Barfoeds Reagent
RCHO + 2Cu2+ + 2H2O RCOOH + Cu2O +
4H+

SUBSTANCES
Benedicts Reagent
Brick-red precipitate
Green to yellow / orange solution

Blue solution
Barfoeds Reagent
Brick-red precipitate

Blue solution


Sodium carbonate: It acts as a catalyst

for releasing enzymes from the

pancreatin so it can break down carbs,
lipids, or protein (mainly starch, though).

WHAT IS THE PURPOSE OF 0.5% SODIUM

CARBONATE SOLUTION IN THE
EXPERIMENT?

It acts as a catalyst for

releasing enzymes from the

pancreatin so it can break
down carbs, lipids, or protein
(mainly starch, though).

HOW DOES SALIVARY AMYLASE COMPARE

WITH PANCREATIC AMYLASE IN ITS ABILITY
TO DIGEST CARBOHYDRATES?
SALIVARY AMYLASE

Chewing breaks food into small molecules that combine with saliva
secreted by the salivary glands in the mouth. Along with mucin and
buffers, saliva contains the enzyme salivary amylase, which acts on the
starch in food and breaks it down to maltose. Salivary amylase continues
for the short duration that the carbohydrates are in the mouth, after which
the mixture of the partially digested carbohydrates travels down the
esophagus into the stomach. Due to the inhibition of salivary amylase
activity by the acidic gastric juices, digestion of carbohydrates does not
occur in the stomach.

PANCREATIC AMYLASE
As the combination of gastric juices and partially

digested food enters the small intestine, the pancreas

secretes pancreatic juices, which contain the enzyme
pancreatic amylase. This enzyme acts on the
remaining polysaccharides and breaks them into
disaccharide units of maltose. In the final step of
complex carbohydrate digestion, the enzyme maltase
present in the lining of the small intestine breaks
maltose into two units of glucose. Glucose is then
absorbed and enters the bloodstream.

PANCREATIC AMYLASE
SALIVARY AMYLASE vs. PANCREATIC AMYLASE
SALIVARY

AMYLASE is produced in the salivary

glands and digestion of simple carbohydrates starts
in the mouth.

PANCREATIC AMYLASE are enzymes produced in the

pancreas, acting more on complex carbohydrates.

PANCREATIC
PROTEASES
Test 2

FUNCTION OF EVERY
COMPONENT USED IN THE
TEST
CuSO4- reduces the property of simple carbohydrates
NaOH-raises the pH of the solution to alkaline
HCl- denatures protein structure; activates

pepsinogen to pepsin

Pepsin- hydrolyzes proteins to smaller polypeptide

and some free amino acids

Pancreatin- a mixture of enzyme from the pancreatin

juice

PURPOSE OF 3N NAOH AND

3N HCL
The protease enzymes produced in the

stomach work best in acidic conditions.

This enzyme is called pepsin. The stomach
produces hydrochloric acid so the enzyme
produced here must be adapted to work
best in these conditions. The optimum pH
for stomach pepsin is approx. pH2

 Pancreatin works best under basic

environment. It has the optimum pH of 7.8

to 8.3.
The pH in the stomach is very acidic,
which would destroy pancreatin unless it is
specifically buffered.

PEPSIN
-an

enzyme that breaks down proteins into smaller

peptides (that is, a protease). It is produced in the
stomach and is one of the main digestive enzymes in
the digestive systems of humans and many other
animals, where it helps digest the proteins in food.

PANCREATIN
Pancreatin contains the pancreatic enzymes
trypsin, amylase and lipase. A similar mixture of
enzymes is sold aspancrelipase, which contains
more active lipase enzyme than pancreatin. The
trypsin found in pancreatin works to hydrolyze
proteins into oligopeptides; amylase hydrolyzes
starches into oligosaccharides and the
disaccharide maltose; and lipase hydrolyzes
triglycerides into fatty acids and glycerols.

Pancreatin is an effective enzyme supplement

for replacing missing pancreatic enzymes, and
aids in the digestion of foods in cases of
pancreatic insufficiency.

PANCREATIC JUICE
a liquid secreted by the pancreas, which contains a

variety of enzymes, including trypsinogen,

chymotrypsinogen, elastase,
carboxypeptidase,pancreaticlipase, nucleases and
amylase.

WHERE ARE THESE

SYNTHESIZED AND WHAT
TRIGGERS THEIR RELEASE?

PEPSIN

Pepsin is most active in acidic environments between 37C

and 42C.Accordingly, its primary site of synthesis and

activity is in the stomach (pH 1.5 to 2). Pepsin will digest up
to 20% of ingested amide bonds by cleaving preferentially
after the N-terminal of aromatic amino acids such as
phenylalanine,tryptophan, andtyrosine.Pepsin exhibits
preferential cleavage for hydrophobic, preferably aromatic,
residues in P1 and P1' positions.

Pepsin'sproenzyme,pepsinogen, is released by the

chief cellsin the stomach wall, and upon mixing with the
hydrochloric acidof thegastric juice, pepsinogen activates to
become pepsin. Pepsin is anaspartic protease, using a
catalytic aspartate in itsactive site.

PANCREATIN
Produced by the exocrine cells of

the pancreas

PANCREATIC JUICE
Pancreatic juice secretion is regulated by the

hormonessecretinandcholecystokinin, which
is produced by the walls of theduodenum
upon detection of acid food, proteins, fats and
vitamins.

WHAT ARE PROTEASES FOUND IN PANCREATIC JUICE?

SPECIFY THE PEPTIDE BONDS BROKEN BY EACH TYPE OF
PROTEASE
Trypsinogen

- precursor form of zymogen of trypsin. It is activated by

enteropeptidase to form trypsin. Trypsin cleaves the
peptide bond on the carboxyl side of basic amino acids
such as arginine and lysine.
Elastase
An enzyme form the class of proteases that break down

elastin

Carboxypeptidase

Protease enzyme that cleaves a peptide

bond at the carboxyl terminal end.
Nuclease

Enzyme capable of cleaving the

phosphodiester bonds between the
nucleotide subunits of nucleic acids

Lipase

Used to break down triglycerides into

free fatty acids and monoglycerides
Chymotrypsinogen
Used to break peptide amide bonds

where the carboxyl side of the amide

bond is large hydrophobic amino acid
(tyrosine, tryptophan and phenylalanine)

Amylase

breaks down starch into

dextrose and maltose

HOW DOES PEPSIN COMPARE WITH PANCREATIC

PROTEASES IN ITS ABILITY TO DIGEST PROTEINS?
Pepsin is an enzyme that breaks down proteins into

smaller particles. In digestion process, these enzymes,

each of which is specialized in severing links between
particular amino acids, collaborate to break down
dietary proteins into their components which can be
readily absorbed by the small intestine. While the
pancreatic protease breaks down most of the types of
proteins. It secreted in the lumen and must be
converted into their active form to digest proteins.