NAME: GROUP NO.
: 6
BOLIVAR, Leonilyn SECTION: BSN1-A
LEGASPI, Ma. Lara Micaela INSTRUCTOR: SAMPOLLO, Leslie
SCORE:
OSUMO, Betiara Jazel
PELINGON, Robert Kris
PIO, Krea Joy
YSULAN, Cyky Mae DATE: December 11, 2020
Module 5: Precipitation, Denaturation and Coagulation
I. MATERIALS
PART A. Protein Denaturation
Test tubes
Beaker
Stirring rod
Water
Egg yolk
Funnel
Ring stand
Erlenmeyer flask
PART B. Methods of Protein Precipitation
Pipet Albumin solution
Acid
Test Tubes
Protein solution
Test Tube Rack
Precipitation of protein with heavy metals
Beaker Lead Acetate Solution
2 mL Acid Solution
Test Tubes
Pipet
Albumin and Picric Acid
Pipet Albumin solution
Picric Acid
Test Tubes
Precipitation of protein by organic solvents
Pipet Albumin solution
Ethanol
Test Tubes
Addition of alcohol to protein solution displaces water molecules
associated in protein thus, decreasing solubility
Pipet Albumin solution
Acetone
Test Tubes
II. PROCEDURES
PART A. Protein Denaturation
Separate in two 150 ml beakers an egg white from the yolk
Add the same volume of water to each and sir it.
Filtrate both solution and use the filtrate as the watery solution of protein to show
their denaturation.
Take a 3 ml of an egg white protein solution placed in a test tube and heat it over
the flame.
Repeat the experiment with the egg yolk solution
Observe the calculation due to the heat.
Concentrated nitric acid add it to an album of solution denaturates
Discards all the solutions in the train. The remained protein solution and the
acidic ones have to be diluted first.
PART B. Methods of Protein Precipitation
Albumin solution
Test tube
Piper
Acid
Protein solution
Precipitation of protein with heavy metals
Using pipet get 2 ml of albumin solution and put it inside the test tube
Add lead acetate solution.
As you can see the addition of lead iron to the protein results in the precipitation
protein as a complex.
Albumin and Picric Acid
Using pipet, get a 2 ml of albumin solution and put it into the test tube.
Transfer 1 ml of picric acid using pipet into the test tube with an albumin solution
inside it.
Observe.
Precipitation of protein by organic solvents
Using pipet, get a 2 ml of protein/albumin solution and put it into the test tube.
Add 2 ml of ethanol also into the test tube
Observe.
Addition of alcohol to protein solution displaces water molecules associated in
protein thus, decreasing solubility
Using pipet, get 2 ml of protein solution and put it into the test tube.
Add 2 ml of acetone into the test tube.
Observe.
III. RESULTS AND DISCUSSION
PART A. Protein Denaturation
EGG WHITE DISCUSSION
Initially, the color was transparent but after
heating the egg firms and the color of the
The egg white becomes solution turns white. Therefore,
HEAT
firm when heated. denaturation does not alter the protein
content and only its secondary and tertiary
structure.
Initially, the color of the egg white was
The egg white turns white transparent but after putting the acetone it
but has a transparent turned into a white color in the top part but
ACETONE
solution at the bottom part the bottom part stays transparent.
of the test tube. Therefore, acetone is also altering the
secondary and tertiary structure of the egg
albumin protein.
Initially, the color of the egg white was
transparent but after putting the
The egg white turns white
concentrated nitric acid, it turned into a
NITRIC at the bottom part and has
white color on the bottom part but the top
ACID a transparent part at the
part stays transparent. Therefore,
top.
concentrated nitric acid also denature the
egg albumin.
PART B. Methods of Protein Precipitation
EGG ALBUMIN DISCUSSION
The egg albumin
turned from Since, the solution turned white or cloudy,
HCl transparent to white we can conclude that denaturation of protein
color after adding the is present.
HCL.
The solution turned
White from Since the solution shows a cloudy material,
LEAD
transparent after we can also conclude that denaturation was
ACETATE
adding the lead present.
acetate.
The egg albumin
does not completely
mix with the picric
Even though the picric acid and egg albumin
acid. A yellow
does not mix totally we can still conclude
PICRIC ACID material was seen on
that picric acid denatures the protein
top and a white or
because a cloudy material is present.
cloudy material was
seen below the yellow
solution.
Same as with picric acid the egg albumin
and 95% ethanol does not mix completely
but we can conclude that denaturation of
Egg albumin also protein is present because a cloudy material
95% ETHANOL
turned cloudy. was seen in the solution. The solution does
not mix because the addition of alcohol to
the protein solution makes protein less
soluble.
POST-LAB QUESTIONS
1. Give the rationale of the use of picric acid for burns and tannic acid for
diarrhea.
Picric acid is used to treat burns because it is astringent and an antiseptic. Picric
acid is a much stronger acid than phenol it decomposes carbonates and may be
titrated with bases. Taking a product containing tannic acid and gelatin “gelatin
tannate” might improve symptoms in children who have had diarrhea for no more
than 3 days. Tannic acid is a naturally occurring plant polyphenol and can be
found in practically all aerial plant tissues. Tannic acid was historically used for
the treatment of diarrhea, topically to dress skin burns and rectally for treatment
of unspecified rectal disorder.
2. Discuss the difference between coagulation and denaturation.
Coagulation and denaturation are two processes that occur in molecules. Both
processes change the state of molecules from native state to a different state.
The difference between coagulation and denaturation is that denaturation is the
changing of the properties of a molecule while the coagulation is the action of
converting the liquid state molecules into the solid or semi-solid state by sticking
molecules together. These are the differences between coagulation and
denaturation.
3. Explain the process of salting out.
Salting out is a purification method that utilizes the reduced solubility of certain
molecules in a solution of very high ionic strength. It is typically, but not limited to,
the precipitation of large biomolecules such as proteins. It occurs in aqueous
solutions of high ionic strength that reduce the molecule’s solubility causing
certain proteins to precipitate. The process of salting out is when the salt
concentration is increased, some of the water molecules are attracted by salt
ions, which decreases the number of water molecules available to interact with
the charged part of the protein. A result of the increased demand for solvent
molecules, the protein-protein interactions are stronger than the solvent solute
interactions, the protein molecules coagulate by forming hydrophobic interactions
with each other.
4. Predict the results in the following tests
COLOR DENATURED COAGULATED
DISCUSSION
REACTION TEST PROTEINS PROTEINS
Initially, the egg
albumin was
colorless, after
adding concentrate
Yellow to orange nitric acid there is
XANTHOPROTEIC Yellow to orange
coloration no change
TEST observed. After
heating the
solution’s color
turned yellow.
Initially, the egg
albumin was
colorless and after
adding the amount
of NaOH solution
and 1% CuSO4
solution there is no
change seen but
when the solution
BIURET TEST Purple color Purple color was heated the
color turned violet
Violet ring In the solution, the
Purple-Violet ring
Violet ring appears indicates
HOPKINS-COLE
the presence of
TEST
Tryptophan.
