Experiment 14 Biochem

LEARNING ACTIVITY
No.14
Protein Digestion
I.OBJECTIVES:
At the end of the experiment, the
students are expected to learn how to:
1. Explain the process of digestion of
protien in food.
2. Write the chemical equation involved
in the digestion of protein.
3. Give the digestive function of pepsin
and trypsin.
4. Described the effect of pH and the
presence of mercury salt on the hydrolysis
of protein.
II.ASSIGNMENT:
1. Arrange the following structure
from most complex to the least;
amino acids, polypeptides, protein.
polypeptides
protein
amino acids
2. Compare the contrast pepsin and trypsin.

What are there digestive functions?
Differences
pH: pepsin is active or shows maximum activity
at acidic pH of 1.5 while trypsin is active at an
basic pH of 7.8
Source:Pepsin is secreted by the mucous lining
of the stomuch region while trypsin is secreted
by the pancreas and helps in digestion of food
in small intestine
Mode of action:pepsin mainly helps in
hydrolysis of proteins while trypsin breaks the
peptide bonds on the carboxylside of selective
amino acids such as lysine and arginine.
Pepsin:
1. It is secreted from the gastric glands of
stomach.
2. It is secreted in inactive form called
pepsinogen.
3. It acts on protein and converts them into
proteoses and peptones.
4. It catalyses in acidic medium.
Trypsin:
1. It is secreted from the exocrine pancreas.
2. It is secreted in inactive form called
trypsinogen.
3. It acts on proteoses and peptones and
converts them into polypeptides.
4. It catalyses in alkaline medium.
The main similarities is that both are

proteases and both are secreted in
an inactive form (zymogen). The
differences you stated are partially
correct: pepsin is a gastric protease
(requiring a strongly acidic pH) and
trypsin is a pacreatic protease
(requiring a slightly basic pH).
Because of their substrate
specificity, they each hydrolyze
specific peptide bonds.
3. Give the effect of pH and the presence of

mercury salt on hydrolysis of protein?
pH: enzyme are sensitive to changes in pH. They
contain groups that can act as acids and bases,
donating or accepting protons, when the pH
change. This can lead to altered protein
recognition. A pH that is very different from
optimum pH can cause denaturation.
Mercury: has a great affinity for a certain minerals,

as well as protein and non-protein molecules in the
body. Have a great affinity of sulfur and sulfhydryl
and are capable of affecting the transsulfuration
pathway in the body
III. DISCUSSION:
The digestion of protien s to

amino acids starts in the
stomach. There the enzyme
pepsin converts proteins to
polypeptides. The conversion is
completed in the small
intestine, where a series of
enzyme breakdown proteins
V.
PROCEDURE:
1.Place a very little piece of hardboiled egg white more or less than
same in size in each of five labeled
test tubes.
2.Add 5 mL of PEPSIN SOLUTION to
the 1st test tube.
3.Add 5 mL of ACIDIFIED PEPSIN
SOLUTION (3.5 mL Pepsin + 1.5 mL
0.7% HCL) to the 2nd test tube.
4. Add 5 mL of BASIC PANCREATIN SOLUTION (3.5

mL Pancreatin + 1.5 mL 0.6% Sodium carbonate)
to the 3rd test tube.
5. Add 5 mL distilled water to the 4th test tube.
6. Add 5 mL of acidified pepsin + mercuric
chloride solution to the 5th test tube.
7. Place the test tube into a warm water bath
(about 40 degree Celsius) and observe for 1 hour.
Record your observations every 10 min-interval
on the data table below.
VI. DATA AND

TIME(mi
RESULTS:
n)
PEPSIN ACIDIFIED
PEPSIN
Observation
BASIC
PANCREATI
N
DISTILLED
WATER
ACIDIFIED PEPSIN
+ Hg Cl2
START
Small
size of
egg
white( n
o
reaction
)
Small size
of egg
white( no
reaction)
Small size
of egg
white
( no
reaction)
Small size
of egg
white
( no
reaction)
Small size of egg

white
( no reaction)
10
The
same
appeara
nce from
the start
Slightly
shrink
The same
appearanc
e from the
start
It swells a
little
The same
appearance from
the start
20
Slightly
shrink
The same
sized(10
mins.) but
it starts
to
dissolve
Slightly
shrink
It swells a
little bit
more
Slightly shrink
TIME(mi
n)
Observation
PEPSIN
ACIDIFIED
PEPSIN
BASIC
PANCREA
TIN
DISTILLED
WATER
ACIDIFIED
PEPSIN + Hg
Cl2
30
It
Continue
It slowly
shrinks to dissolve dissolved
a little
slowly
and it
dissolve
a little
It swells a
little
It shrinks a
little
40
it
Continue
dissolve to dissolve
a little
slowly
Same
appearan
ce (30
mins.)
It becomes
bigger
compare to
the
previous
observation
Same size (30

mins.) but it
dissolve a
little
50
Same
It becomes it dissolve
appeara
slightly
a little
nce
cloudy in
(40mins appearanc
.)
e
It becomes
bigger and
does not
disolved
Continue to
dissolve
slowly
TIME(mi
n)
Observation
PEPSIN
60(1 Hr)
ACIDIFIED
PEPSIN
BASIC
PANCRE
ATIN
it
Continue Continue
dissolve to dissolve
to
a little
slowly
dissolve
slowly
DISTILLED
WATER
ACIDIFIED
PEPSIN + Hg
Cl2
Same
appearance
(50mins.)
Continue to
dissolve
slowly
VII. Conclusion:
Based on the experiment we
preformed, we therefore conclude
that most proteins are
decomposed to single amino acids
in digestion.
Digestion typically begins in the
stomach when pepsinogen is
converted to pepsin by the action
of hydrochloric acid. Hydrochloric
acid is used for breaking the
bonds between the proteins.
In the different test tubes we

observed that in:
Tube 1: thick solid white mass,
partially digested
Tube 2 - white cloudy liquid but
have small portion of solid,
extensively digested
Tube 3 - very small white
molecules settled down, very
little digestion
Tube 4 - white solid mass, not
digested
Tube 1 is only partially

digested because it also
needs HCl to complete
protein digestion.
Tube 2 is digested because
it is the ideal environment
of the stomach
Tube 3 shows little
digestion because sodium
carbonate is used and acts
Tube 4 is not digested

because it needs pepsin to
turn protein into amino
acids
tube 5 shows little
digestion because Mercuric
chloride.
POST LABORATORY
QUESTIONS:
Can the body store carbohydrates? Fat?

Protein? If so, in
what form?
Yes, our body has a limited storage capacity

for carbohydrates in the muscle glycogen
and the liver glycogen. When those supplies
are full, excess carbohydrates are stored as
fat. The body stores carbohydrate as
glycogen in the muscles and liver, however
its storage capacity is limited.
When you consume more carbohydrates
than you immediately need, your body
stores them as a substance called
glycogen.
The body does not store protein, rather it

takes what it needs from both the amino
acid pool and what is derived from food
sources and uses it for the necessary
functions mentioned. When more protein
is consumed than what the body needs,
the protein is processed through the liver,
filtered through the kidney and then
excreted through urine. For this reason,
Berardi recommends that women need
only to consume 20 to 30 g of protein at a
time, and for men, 40 to 60 g is sufficient.

When you eat more fats than you
immediately need, your body stores them
as a substance called triglyceride, or
triacylglycerol.
When you eat more fat than you need for
your short-term function, fatty acids from
your bloodstream combine inside your fat
cells with a substance called glycerol
and form triglyceride. When you need to
draw on your fat reserves, triglyceride is
broken apart and sent back into your
What is the normal daily requirement of

protein? What can affect this amount?
Protein is the essential nutrients for the

human body. They are one of the building
blocks of body tissue, and can also serve
as a fuel source. As fuel, proteins contain
4kcalper
gram,
just
likecarbohydratesand
unlikelipids,
which contain 9 kcal per gram.
Proteinsarepolymerchains
made
ofamino acidslinked together bypeptide
bonds.
According to US & CanadianDietary Reference

Intakeguidelines, women aged 1970 need to consume
46grams of protein per day, while men aged 1970
need to consume 56grams of protein per day to avoid a
deficiency. The American and Canadian guidelines
recommend a daily protein dietary allowance, measured
as intake per kilogram body weight, is 0.8 g/kg.
Protein requirements are complicated because the
amount we need changes with age.
Infants require about 10 grams a day.
Teenage boys need up to 52 grams a day.
Teenage girls need 46 grams a day.
Adult men need about 56 grams a day.
Adult women need about 46 grams a day.
Protein is found in the following foods: meats,

poultry, and fish, legumes (dry beans and peas),
tofu, eggs, nuts and seeds, milk and milk
products, grains, some vegetables, and some
fruits (provide only small amounts of protein
relative to other sources).
When a high dietary protein intake is consumed,
there is an increase in urea excretion, which
suggests that amino acid oxidation is increased.
Unlike fat and glucose, our body has little
capacity to store protein. If we were to stop
eating protein, our body would start to break
down muscle for its needs within a day or so.
THE
END
GROUP 6:
Tahad, Jojean
Taghap, Jesil
Tormis, Lady
Lee
Ramos, Maria

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LEARNING ACTIVITY

2. Compare the contrast pepsin and trypsin.

The main similarities is that both are

3. Give the effect of pH and the presence of

Mercury: has a great affinity for a certain minerals,

The digestion of protien s to

4. Add 5 mL of BASIC PANCREATIN SOLUTION (3.5

VI. DATA AND

Small size of egg

Same size (30

In the different test tubes we

Tube 1 is only partially

Tube 4 is not digested

Can the body store carbohydrates? Fat?

Yes, our body has a limited storage capacity

The body does not store protein, rather it

What is the normal daily requirement of

Protein is the essential nutrients for the

According to US & CanadianDietary Reference

Protein is found in the following foods: meats,

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