ENZYMES AND

DIGESTION

Mark Anthony Batitis

Jed Riel Bautista
Mariella Kaycee Caluag
Louise Michaela Cerdea
1NUR3 Group 2
Introduction
 Enzymes
Cells produce special proteins
Are important molecules, they are particularly important in
digestion as they help break down large, insoluble food
molecules into small, soluble molecules which can be easily
absorbed.
Catalyzes biological reactions by lowering the activation
energy of reaction
Subsrate a reactant broken down by an enzyme
Are either simple proteins or protein (apoenzyme) combined
with a non-protein unit (cofactor)
 Enzymes and cells
Enzymes catalyze the thousands of reactions that need to take place in order to maintain
life. What are some of these reactions?

digestion

respiration

photosynthesis (plants and some bacteria)

protein synthesis.
 Functional Unit of
Enzymes
Coenzyme cofactor is an organic unit
Metal-ion activator cofactor is metal-ion
The two parts (protein & non-protein)
constitutes a Haloenzyme
FUNCTION OF
ENZYMES
Their basic function is to speed up the process and efficiency of a
reaction without themselves being consumed in the process.

Enzymes are responsible for moving large parts of a cells internal

structure, such as pulling chromosomes apart when a cell divides.
Enzymes make the energy molecules that are constantly needed for
the cell to survive.

And they break down molecules, recycle the old parts and make new
molecules that allow the cell to grow.
 What are digestive
enzymes?
Not all enzymes work inside cells. In what process do enzymes work outside cells?

Digestive enzymes are produced by specialized cells in the pancreas and digestive tract.

These enzymes pass out of the

cells and into the stomach and
small intestine.

Here the enzymes help to break down

large food molecules into smaller
molecules that are more easily
absorbed.
Digestion in the stomach
When food enters the stomach it stimulates the secretion of hydrochloric (HCl) acid from
the stomach wall.
Hydrochloric acid increases the acidity of the stomach to about pH2. This is the optimum pH
for stomach enzymes.
oesophagus

mucus cells

gastric gland

parietal cells
(acid-producing)

duodenum
 Digestion in the small intestine
Digestive enzymes found in the small intestine are damaged by a strongly acidic pH.
How does the body avoid this problem?

The liver produces bile (an alkali), which is stored in the gall bladder and released into the
small intestine.

Bile neutralizes the acidic contents hepatic

coming from the stomach, creating duct
the alkaline environment that the
intestinal enzymes need to work. gall
bladder

pancreas

duodenum bile duct

 Lock and Key Model
Each type of enzyme catalyses a particular reaction or type
of reaction because enzymes only work for a specific
substrate. Which is the enzyme is the lock and the specific
substrates is the key. They can do this because the long
chains of amino acids which make up the enzyme are folded
into a particular shape known as theactive site. The
substrates of the reaction fit into the active site of the
enzyme. This works like a lock and key. Once the substrate
is in place in the active site it binds to the enzyme and the
reaction then takes place rapidly. The products of the
reaction are released from the surface of the enzyme.
Lock and Key Model
What is a substrate?

a substrate is typically the chemical

species being observed in a chemical
reaction, which reacts with reagent to
generate new product.
How do enzymes act on substrate?

In the lock-and-key model, the active site of an enzyme is

precisely shaped to hold specific substrates. In the induced-fit
model, the active site and substrate don't fit perfectly together;
instead, they both alter their shape to connect.

Whatever the case, the reactions that occur accelerate greatly

over a millionfold once the substrates bind to the active site of
the enzyme. The chemical reactions result in a new product or
molecule that then separates from the enzyme, which goes on to
catalyze other reactions.
4) How can one classify enyzmes?
Metabolism influences building or replacement of tissue, conversion of food to
energy, disposal of waste materials, reproduction etc. Catalysis is defined as
the acceleration of a chemical reaction by a substance which itself undergoes
no permanent chemical change. Most biochemical reactions do not take place
spontaneously and enzyme catalysis plays an important role in biochemical
reactions necessary for all life processes. Without enzymes, these reactions
would take place at a rate far too slow for effective metabolism.

Enzymes can be classified by the kind of chemical reaction they catalyze. One
such scheme of enzyme classification is defined by IUBMB.

The IUBMB assigns a 4-digit code to each enzyme. Each enzyme is prefixed by
EC, followed by the digits.
 1. The first digit denotes Class of the enzyme

2. The second digit indicates, Sub-class of the

enzyme

3. The third digit gives Sub sub-class of the enzyme

4. The fourth digit in the code is Serial number of

the enzyme
Group Name Type of Example
ReactionCatalysed
Oxidoreductases Oxidation-reduction Alcoholoxidoreductase(E
reactions C 1.1)
Transferases Transfer of functional Methyltransferase(EC
groups 2.1)
Hydrolases Hydrolysis reactions Lipase (EC 3.1)
Lyases Addition to double bonds Decarboxylases(EC 4.1)
or single bonds

Isomerases Isomerizationreactions EpimerasesandRacemas

es(EC 5.1)

Ligases Formation of bonds with Enzymes forming carbon-

ATP cleavage oxygen bonds (EC 6.1)
Digestion
In order for the nutrients in food to be absorbed, they
must first be broken down into particles that are small
enough to be transported through carrier proteins into
the epithelial cells that form the mucosal lining of the
digestive tract.
Occurs primarily within three particular segments of the
digestive tract: the mouth, the stomach, and the small
intestine.
Physical Digestion
large chunks of food are ground into tiny particles
Chemical Digestion
whereby through the use of enzymes released into the
digestive tract large polymeric biomolecules are broken
into individual monomers or oligomers (e.g. dimers or
trimers).
Digestive enzymes greatly enhance the rate at which
the covalent bonds that link subunits together to form
polymeric biomolecules are broken.
ENZYMES AND
DIGESTION
A. FACTORS AFFECTING ENZYMATIC
ACTIVITY
 1. Effect of Temperature
Temperature can effect an enzyme in two ways. One is a direct
influence on the reaction rate constant, and the other is in thermal
denturization of the enzyme at elevated temperatures.

To relate the effect of temperature to the reaction rate constant,

the Arrhenius equation is used:
Temperature affects thereaction rate of enzymes, as do pH, substrate concentration
and enzyme concentration. Atlow temperatures, enzymes havelow activity. As the
temperature rises therate of reaction increases, usually2-fold for every 10 degree
Celsius rise.
Optimum temperature is the temperature at which the enzyme has it maximum activity
(highest rate of reaction) Beyond the optimum temperature the activity of the enzyme
decreases. At extreme temperatures, the enzymes aredenaturedand activity ceases.
Since pancreatic amylase is a digestive enzyme its usual
optimumtemperature would be body temperature ie.37oC
All reactions are faster at a higher temperature. However, enzyme-catalyzed
reactions become slower or stop if the temperature becomes too high, because
enzymes become denatured at high temperatures .
STARCH IS THE SUBSTRATE. SUBSTRATE OF BOTH
TEST
 RANKING OF THE TUBES

2 1
Dark Blue Presence of Starch
Violet or red Partial digestion of starch
Colorless Complete digestion

370 BOILING W.B. ICE W.B.

Observation & contrast of different
Temp.
Test tube B came through the process of 370 C where
digestion came up faster among the 2 tubes
Test tube C was the slowest among them maybe due to the
temperature and acidity.
 How can iodine determine the degree
of digestion in the given tubes?

Used to test for the presence or absence of starch in the solutions using
iodine (I2). Iodine forms a blue to black complex with starch, but does not
react with glucose.
*SAME ANSWERS OF BOTH TESTS*
 If iodine is added to a glucose solution, the only color seen is the
red or yellow color of the iodine. Therefore, the faster the blue color
of starch is lost, the faster the enzyme amylase is working. If the
amylase is inactivated, it can no longer hydrolyze starch, so the
blue color of the starch-iodine complex will persist.
 Relevance to Human
health
Enzymes are proteins with three-dimensional shapes that define their function. The active site is
the most crucial part of the enzyme and is akin to a mouth with teeth that are properly in place.
The atoms making up protein molecules can vibrate to different degrees depending on the
temperature.
Enzymes have comfort zones, outside of which they stop working. Boiling or freezing an enzyme
completely stops its activity. However, it is interesting to note that what would be considered as
boiling and freezing temperatures that are inhospitable to human life still have enzymes that
function normally therein.
Relevance to Human health
Too much jiggling of enzyme,
due to boiling
Too little movement of
enzyme, due to freezing, will
slow down an enzymes
activity
 2. Effect of pH
Most proteins, and therefore enzymes, are active only within a narrow pH range usually
between 5 and 9. Several factors are influenced directly by the pH in which the reaction takes
place.

the binding of substrate to the enzyme

the ionization states of the amino acid residues involved in the catalytic activity of the
enzyme.
the ionization of the substrate
variation in the protein structure at extreme pH.
Each enzyme has an optimum pH. Above or below an enzymes
optimum pH, its activity is lower. The optimum pH of a particular
enzyme corresponds to the pH of its natural environment. For many
enzymes, this corresponds to pH values of around 7. (check the
relevance in health category)
STARCH IS THE SUBSTRATE. SUBSTRATE OF BOTH
TEST
 REMEMBER:
Both of tests requires fresh amylase.
A fresh amylase solution by collecting about 1-2 mL of your
own saliva in a small beaker or vial but there are diff. types of
amylase and this depends on the persons amylase that may
cause failure or success throughout the reaction of this test!
Most failed in this test. I also failed at the ranking of these pH.
RANKING OF THE TUBES

3 1 2
2 1
Dark Blue Presence of Starch
Violet or red Partial digestion of starch
Colorless Complete digestion

pH 4 pH 7 pH 10
Observation & contrast of different
pHs
pH of 4 wasn't clear at the end of 10 minutes.
pH of 7 took about 10 minutes to turn clear.
pH of 10 took about 7 minutes to turn clear.
 Hypothesis of this test
The neutral pH's would turn clear first because the extreme acidic or
extreme basic would effect the enzyme negatively by denaturing it.
We did not put into consideration the optimal pH of the amylase of the
most acidic acid that would break down the starch into sugars. Our
hypothesis was not supported due to the fact that the pH of 10 turned
clear first and the neutral pH's of 7.
 How can iodine determine the degree
of digestion in the given tubes?

Used to test for the presence or absence of starch in the solutions using
iodine (I2). Iodine forms a blue to black complex with starch, but does not
react with glucose.
*SAME ANSWERS OF BOTH TESTS*
 If iodine is added to a glucose solution, the only color seen is the
red or yellow color of the iodine. Therefore, the faster the blue color
of starch is lost, the faster the enzyme amylase is working. If the
amylase is inactivated, it can no longer hydrolyze starch, so the
blue color of the starch-iodine complex will persist.
 Relevance to Human
health
Components in saliva help keep the pH in your mouth between 6.5 and 7 so
that the enzyme salivary amylase can start to break down carbohydrates.
The enzymes that help digest food in the stomach, such as pepsin, work
best at a pH around 2, while those that function in the intestines, including
peptidases and maltase, work best at a pH around 7.5
Effect of Enzyme Concentration

REPORTER: Jed Riel D. Bautsita 1 NUR 3

 Control
standard experimental
treatment which
includes all the
factors thought to be
the rate of a chemical reaction is affected by the
significant, exept one, total number of enzymes

so that details of its

effects may be
discovered by
comparison.
RESULTS
Effect of Enzyme Concentration

The 'lock and key' hypothesis explains how

enzymes only work with a specific substrate.
The hypothesis presents the enzyme as the
'lock, and the specific substrate as 'key'. The
active site binds the substrate, forms a
product, which is then released. The more the
enzyme concentration it will increase the rate
of reaction, as more enzymes will be colliding
with the substrate which is the starch.
PRINCIPLE
how can Iodine solution determine the degree of
digestion?

Answer:
using the chemical reaction of Iodine to identify the product or reaction of salavi
with the starch using its color reaction. Iodine solution react with start a bluish
color is obtained indicating the presence of starch.Some polysaccharides have have
the property of adsorption for iodine.So, they adsorb iodine and give coloration.
HEALTH IMPLICATIONS
Starch is a carbohydrate onsisting of a large number of
glucose units joined by glycosidic bonds. this
polysaccharide is produced by most green plants as an
energy store. It is the most common carbohydrate in
human diets and is contained in large amounts in staple
foods. It is the main carbohydrrate in human nutrition.
 SUBSTRATE
CONCENTRATION
Results: negative

The positive result should be brick red solution is

a + results which means that the starch was
fully digested. The blue color sol. is a negative
result.Sugar needs to be decomposed into its
components glucose and fructose then the
glucose test would be positive but the starch
test would still be negative.
 Substrate concentrations affect the rate of reaction of an enzyme-
catalysed reaction. Controlling these factors in a cell is one way that
an organism regulates its enzyme activity and so its Metabolism.
Changing the concentration of a substance only affects the rate of
reaction if it is the limiting factor: that is, it the factor that is
stopping a reaction from preceding at a higher rate.
If it is the limiting factor, increasing concentration will increase the
rate of reaction up to a point, after which any increase will not affect
the rate of reaction. This is because it will no longer be the limiting
factor and another factor will be limiting the maximum rate of reaction.
As a reaction proceeds, the rate of reaction will decrease, since the
Substrate will get used up. The highest rate of reaction, known as the
Initial Reaction Rate is the maximum reaction rate for an enzyme in an
experimental situation.
Benedicts Principle:
When Benedicts solution and simple
carbohydrates are heated, the solution
changes to orange red/ brick red. This
reaction is caused by the reducing
property of simple carbohydrates. The
copper (II) ions in the Benedicts
solution are reduced to Copper (I) ions,
which causes the color change.

Reducing sugars are oxidized by the

copper ion in solution to form a
carboxylic acid and a reddish
precipitate of copper (I) oxide.
Effect of Metal-ion
Poisons on Enzyme
Activity
 Purpose:
To determine how metal ions affect
enzyme activity.
Reagents:
Lead Acetate, Silver Nitrate
Principle:
Denaturation of enzymes by the heavy
metals
Metal-Ion

Enzyme:
Amylase (Salivary)
Substrate:
Starch
Manipulated Variable:
Heavy Metal-ion
Procedures:
Prepare 3 clean and dry test tubes and label them
from 1 to 3. Place 1.0 mL of saliva into each of the
tubes.
To the first tube, add 3.0 mL of distilled water 3.0
mL of lead acetate solution to tube 2; and 3.0 mL of
1% silver nitrate solution to tube 3. Mix thoroughly.
Place all tubes in the water bath set at 37C for 10
minutes. Remove the tubes from the water bath at
the same time.
Prepare 3 clean and dry vials and label as 4 to 6. put
3.0 mL of 1% starch solution into each of the vials.
Procedures:
Transfer 10 drops of the solutions into the vials: tube
1 to vial 4, tube 2 to vial 5 and tube 3 to vial 6.
incubate at room temperature for 10 minutes while
periodically mixing the contents of the vial.
Record the color of the solutions and indicate if
digestion took place
With Complete digestion ------------------- Colorless
With Partial digestion --------------------- Violet or
Red
No digestion------------------------------------Dark shade
of Blue
Heavy Metal Ion
Heavy Metal ions affect the activity of
the amylase (Enzyme). It binds to the
enzyme, outside the active site in the
different place then substrate. The
enzyme active site binds substrate but
cannot convert it into a product.
Heavy metal salts act to denature
proteins in much the same manner as
What is the Health implication of this procedure?

This reaction is used in reverse

in cases of acute heavy metal
poisoning. In such a situation, a
person may have swallowed a
significant quantity of a heavy
metal salt.
What is the Health implication of this procedure?

As an antidote, a protein such as

milk or egg whites may be
administered to precipitate the
poisonous salt. Then an emetic
is given to induce vomiting so
that the precipitated metal
protein is discharged from the
Enzyme Inhibitors
Enzyme inhibitors are molecules that
interact in some way with the enzyme to
prevent it from working in the normal
manner.
It is the prevention of an enzymic process
as a result of the interaction of some
substances with the enzyme so as to
decrease the rate of the reaction.

There are 2 type of inhibitions:

Heavy Metal as an inhibitor
Heavy Metals are noncompetitive Inhibitor.
Non-Competative inhibitors are substances
that do not bind to the active site of an
enzyme, but rather to the other parts of
the enzyme. In doing so, they may change
the conformation of the active site as has
already been explained.
Contents VIAL 4 VIAL 5 VIAL 6
SALIVA
Starch

Solution
Distilled Lead Silver
REAGENTS
Water Acetate Nitrate
Pb(C2H3O2)
( H20 ) AgNO3
2
Pancreatic
Amylase
 Purpose:
To determine what kind of enzyme from
the pancreas digest starches
To differentiate salivary amylase from
pancreatic amylase
Reagents:
Benedicts Reagent, Barfoeds Reagent
Principle:
Hydrolysis by Amylase, Oxidation of Sugar
Pancreatic Amylase
Enzyme:
Salivary Amylase
Pancreatic Amylase
Substrate:
Starch
Manipulated Variable:
Barfoeds and Benedicts Reagent
Procedures:
Put 1.0 mL of 1% starch on each of 2
clean and dry test tubes.
To the first tube, add 1.0mL of saliva.
And to tube 2, add 1.0mL of 5%
pancreatin and 1.0 mL 0.5% Sodium
Carbonate solution. Mix thoroughly
and place in a water bath set at 37C
for 10 minutes.
Procedures:
To the first portion, add 3.0 mL of
Benedicts Reagent and warm in a
boiling water bath for 5 minutes. Note
the color of the resulting solution and
precipitate, if any.
Record all observations. A brick-red
precipitate as well as a green to
yellow or to an orange colored
Procedures:
To the first portion, add 3.0 mL of
Barfoeds Reagent and warm in a
boiling water bath for 2 minutes. Note
the color of the resulting solution and
precipitate, if any.
Record all observations. A brick-red
precipitate is a positive result. A blue
solution is a negative result.
Pancreatic Amylase
Pancreatic Amylase is an important
metabolic enzyme. Its function is to
catalyze the hydrolysis of starch into
glucose. This particular enzyme which
is found in all mammals speeds up
digestive processes which take place
along the digestive track running from
the mouth to the small intestines.
What is the principle behind the use
of Benedicts and Barfoeds Test?
Benedicts test - to detect
presence of reducing sugars
Barfoeds test - to detect
presence of
monosaccharides
What is the principle behind the use
of Benedicts and Barfoeds Test?
In Benedicts Test, it is a test to
determine a reducing sugar. Since the
salivary amylase can break down
starch, a polysaccharide, into
disaccharides, a maltose which is a
composition of two glucose, therefore a
reducing sugar, it will be detected by
the Benedicts Reagent, and therefore,
would precipitate.
What is the principle behind the use
of Benedicts and Barfoeds Test?
In Barfoeds Test, since it is a test to
determine a reducing monosaccharide
and since the pancreatic amylase can
break down starch, a polysaccharide,
into monosaccharides, glucose, and
the sodium carbonate acted as a
catalyst in the process, it will be
detected by the Barfoeds Reagent,
therefore, would precipitate.
Salivary Amylase
- an enzyme made by salivary
gland or found in saliva
- acts on starch to break it into
many molecules of maltose.
Discussion of results
 Pancreatic Amylase:
- produced by pancreas
- acts on starch to convert it to maltose
that occurs in duodenum (first part of the
small intestine immediately beyond the
stomach).
- during digestion, starch is partially
transformed into maltose by the pancreatic
or salivary enzymes called amylases;
RESULTS
Sodium carbonate
acts as a catalyst
for releasing
enzymes from the
pancreatin to
break down
carbohydrates,
lipids, or proteins
(mainly starch).
 Test Tube Benedicts Test Barfoeds Test
Blue solution (negative)
Yellow-green
- negative because of
precipitate (positive)
presence of
Saliva - positive of presence
disaccharide (maltose)
of reducing sugar
and not further
(maltose)
digestion of maltose
Blue solution (negative)
Yellow precipitate
- Negative because of
Pancreatin (positive)
presence of
and Sodium - positive because of
disaccharide (maltose)
Carbonate presence of reducing
further digestion of
sugar (glucose)
maltose
SUMMARY
Salivary amylase
- enzymes produced in the salivary glands
- gets mixed with the other components of saliva when
food is chewed in the mouth
- partial digestion of carbohydrates is already ongoing in
the mouth even before food goes into the stomach and small
intestines

Pancreatic amylases
- enzymes produced in the pancreas
- pancreas is the main digestive organ, producing powerful
 PANCREATIC
PROTEASES
Louise Michaela Cerdea (Group 2)
Objectives
To describe the role of enzymes in the digestive
process
To be able to identify the tests used in the
experiment and its principles
To be able to identify the products of digestion of
proteins in the digestive tract.
ProCedure
a. Prepare 3 clean and dry test tubes and transfer the
following solutions as follows:
SOLUTION TEST TUBE 1 TEST TUBE 2 TEST TUBE 3
Egg White Solution 2.0 mL 2.0 mL 2.0 mL
CuSo4, 1% 5 drops 5 drops 5 drops
3N NaOH 2.0 mL 2.0 mL Add until basic to
litmus
3N HCl - Add until acidic to -
litmus
Pepsin - 2.0 mL -
Pancreatin - - 2.0 mL
b. Add the pepsin and
pancreatin simultaneously.
Quickly mix the contents of
all tubes thoroughly.
c. Immerse all tubes in a water
bath set at 37C until a
change in color of the
solutions is observed
d. Remove all tubes from the
water bath at the same time.
Compare the intensity of the
colored solutions produced.
Record all your observations.
RESULTS
TEST TUBE

(1) Violet(No
No enzyme Digestion
present
(2) Light Brown
Pepsin with white
precipitate(Digestio
n)

(3) Brown(Digestion
Pancreatin )
Discussion

Proteases, also called proteinases or peptidases are

important enzymes that function by breaking proteins
into smaller peptides. Proteases occur in a number of
organs in the gastrointestinal tract and are essential
for absorption of peptides by the intestinal mucosa.
The digestion of proteins is initiated by Pepsin in the
stomach, but the bulk of the protein digestion is due
to the pancreatic proteases.
Biuret Test is used to determine the presence
ofpeptide bondsin the protein which is based on the
ability of Cu (II) ions to form a violet-coloured chelate
complexwith peptide bonds in alkaline conditions. The
greater theconcentrationof peptide bonds, the greater
thecolor intensity. Copper(II) Sulfate (CuSO4 1%) and
Sodium Hydroxide (3N NaOH) is used as the reagent.
Reduction and Complexation
(Test Tube 1)
The two most important for initiating protein digestion
are Hydrochloric acid (HCl) and the protease
Pepsin.

HCl is secreted by parietal cells in the gastric mucosa.

The low pH (~2) of the gastric juice aids protein
digestion in 2 ways:
First, the low pH denatures the tertiary structures of
ingested protein, making them easier to digest
enzymatically.
Secondly, the low pH is required for the activation of
pepsin.
Pepsin is produced by chief cells of
the gastric mucosa in an inactive
(zymogenic) form called
pepsinogen. Pepsinogen is inactive
when released in the gastric pits,
but once it diffuses into the lumen
of the stomach the acidic conditions
enable it to have a weak proteolytic Hydrolysis of Amide Linkages
activity.

Denaturation Hydrolysis by Pepsin

What is Pancreatic Juice?
Pancreatic juiceis a liquid secreted by thepancreas,
which contains a variety ofenzymes. These juices
consist primarily of water, NaCl (salt), and NaHCO3
(sodium bicarbonate). The purpose of the sodium
bicarbonate is to neutralize the high acidity of the
chyme (food plus stomach acid) raising it to an
alkaline pH of 7.1-8.2. This both stops the action of
gastric pepsins and stomach acid and prepares
chyme for the process of nutrient absorption, which
takes place in the small intestine. Pancreatin contains
the enzyme pancreatic protease that aids in the
digestion of protein.
The proteases found in the pancreatic juice,
trypsin and chymotrypsin, are
endopeptidases, or enzymes that attack internal
peptide bonds. These two pancreatic digestive
enzymes work together and, with aid from the
pepsins located in the stomach, they are
responsible for beginning the digestive process.
Pepsin Pepsin is a digestive enzyme found in
gastric juice that catalyzes the breakdown of
protein to peptides. Pepsin is one of the
three principal protein-degrading, or
proteolytic enzymes in the digestive system.
It degrades food proteins in the stomach
and acts as a potent proteolytic enzyme
cleaving proteins into peptides in the gastric
lumen at a low pH.
Chymotrypsin Chymotrypsin is a pancreatic digestive
enzyme that catalyzes the hydrolysis of
certain proteins in the small intestine into
polypeptides and amino acids.
Chymotrypsin hydrolyzes the peptide bond
of amino acids with large hydrophobic side
chains, such as phenylalanine, tryptophan,
and tyrosine.
Trypsin Trypsin is a proteolytic enzyme that
hydrolyzes peptide bonds on the carboxyl
side of the amino acids arginine and lysine.
Trypsin catalyzes the cleavage and
PANCREATIC LIPASE
AND BILE
Objectives
To describe the role of enzymes in the digestive
process
To be able to identify the tests and principles
used in the experiment
To be able to identify the products of digestion of
fats in the digestive tract.
 Procedure
a. Prepare 3 clean dry test tubes and transfer the
contents as follows:
Content Test Test Test
s Tube 1 Tube 2 Tube 3
Vegetable 20 drops 20 drops 20 drops
Oil
Pancreati 3.0mL - 3.0mL
n
Sodium - 3.0mL 3.0mL
Choleate
Distilled 3.0mL 3.0mL -
Water
b. Mix the contents of all tubes thoroughly.
c. Note the initial pH using pH paper. Record the
initial pH. Adjust the initial pH to 7 (or a little bit
higher) by adding 0.1% NaOH dropwise.
d. Immerse all the tubes in a water bath set at 37C
for 1 hour. Check the pH of your solutions every 15
minutes. Removes all tubes from the water bath at
the same time. Record the final pH.
 RESULTS
Test Tube 1 Test Tube 2 Test Tube 3

Initial pH 11 49 11

Final pH 11 10 11
Discussion

Pancreatic Lipase(found in Pancreatin) is a

water-soluble enzyme secreted by the
pancreas. Like other lipases, its function is to
break down lipids (fats) in the intestinal tract.
However, unlike other pancreatic enzymes such
as trypsinogen and chymotrypsin, pancreatic
lipase is secreted in pancreatic juice as an
active enzyme and doesnt need to undergo
conversion to digest lipids.
 This enzyme possesses the ability to break down
dietary fats via hydrolysis by breaking the ester
linkages. One of the primary tasks of pancreatic
lipase is to break down triglycerides(vegetable
oil) into a2-monoglyceride and two free fatty
acids in alkaline condition. This is critical since
these particular lipids cannot be absorbed
through the intestinal lining without first
undergoing hydrolysis.
Lipids are insoluble while lipase is soluble in water. Thus,
lipase cannot break down lipids directly. Hence an emulsifier
(bile salt---sodium choleate) is essential. Bile is a digestive
fluid that is synthesized in the liver. Bile salts are
amphipathic. By making the lipids soluble in water, bile salts
enable lipids to be digested
There are two fundamentally important functions of bile in
all species:
(1) Bile contains bile acids, which are critical for digestion
and absorption of fats and fat-soluble vitamins in the small
intestine
(2) Many waste products, including bilirubin, are eliminated
from the body by secretion into bile and elimination in feces.
Correct results
Initial pH 7 7 8

Final pH 7 7 6

Since lipids contain fatty acids, when

hydrolyzed, there should be a
decrease in pH.