Name: BATAC, JOSHUA KYLE C.

Date: __________________
Group: ______________________________________ Rating:_________________

PHINMA UNIVERSITY OF PANGASINAN

COLLEGE OF ALLIED HEALTH SCIENCES
DEPARTMENT OF NURSING

ACTIVITY NO. 6
DENATURATIONS OF PROTEINS

I. DISCUSSION:
Nature / Organization of Protein Molecule
Properties of Proteins

II. PROCEDURE:

A. COAGULATION OFPROTEINS

1. By Heat:
Place 3 ml. of egg albumin solution in a test tube and boil gently for a few
minutes. Describe the change in the albumin solution.
Result: When heated, the protein in egg whites solidified. Additionally, it becomes
white and coagulates, indicating that protein denaturation occurred but did not affect the
protein's primary or secondary structure. The heat causes coagulation in the yellow yolk as
well.

2. Heller’s Ring Test

Place 1 ml. of conc. nitric acid in a test tube (CAUTION!). Dilute 1 ml. of egg albumin
solution with q ml. of water and mix well. Incline the tube containing the nitric acid and
carefully pour the 3 ml. of diluted albumin solution down the side of the tube so that it
forms a layer above the acid solution. What do you observe at the juncture of the two
liquids?
Result: The egg albumin is denaturized by the additional concentrated nitric acid. The
albumin changes to a murky hue.

3. By acetone
Place 1 ml. of egg albumin in a test u\tube and add 1 ml acetone . What changes
occurs in the albumin solution?
Result: Acetone addition reduces the amount of water that is available in proteins,
thereby reducing their solubility. The egg albumin changes in the albumin solution, but only
at the top of the solution does it take on a white, foggy color. Acetone also has the ability to
denature proteins.
B. PRECIPITATION OF PROTEINS

1. By strong Acid (CAUTION!)

In each of 2 test tubes containing 3 ml. of clear 1% egg white solution, add 2ml of
hydrochloric acid . Note and describe the changes that occur.
Result: The formation of white, cloudy material indicates denaturation.

2. By Heavy Metal Cations

In each of 2 test tubes containing 2ml.of egg white solution, add several drops of5% lead
acetate solution.
Result: The precipitation of protein as a complex is caused by the addition of lead ion
to the proteins. A complex white-clouded solution was also produced.

3. By Alkaloidal Reagent
To 1 ml. o 1% egg white solution add 2 ml of saturated aqueous picric acid
Result: A cloudy substance is proof that picric acid has denaturized the protein.

Why is picric acid used in the treatment of burns?

Answer: By altering their tertiary structure, proteins can be denatured by picric acid.
By altering the H-bonding and Vander Waals dispersion forces, picric acid can precipitate
protein particles so that the hydrophilic end of the protein is in the middle, where picric acid is
normally bound, and the hydrophobic end is in the outer stratum. Your denatured protein
consequently precipitates as an insoluble precipitate. In a sense, obstructs the path for any
infection or bodily fluid to leave or enter the wound, including interstitial fluid. These support
your body's fibrinogens in their fight against infection and enable it to keep a healthy number
of monocytes in the affected area. Additionally, picric acid has an antiseptic quality that can
be used to treat wounds.

4. By organic solvent
To 2 ml of 0.1% egg white solution add few drops of ethanol.
Result: Alcohol increases the solubility of proteins by displacing water molecules in
protein solutions.

III. QUESTION:

1.Explain how proteins become denatured.

A protein becomes denatured when some of the hydrogen bonds in its normal structure
are broken. Weak hydrogen bonds can break when too much heat is applied to them or when
they are exposed to an acid. Proteins interact with one another as they unravel or deform,
forming interactions with other protein molecules that cause them to coagulate and become
insoluble in water.

2. What is meant by isoelectric point? What is its importance?

The isoelectric point (pI) is the pH at which a molecule carries no net electrical charge.
The pH of the environment around the molecule affects its net charge, which can change
when protons are gained or lost to become more positive or negative. The isoelectric point is
significant for protein purification because it indicates the pH where solubility is typically
lowest. In an electro-focusing system, the protein will gather at the isoelectric point, which is
the location where mobility is zero.

3. What is the protein in milk? Why is this protein precipitated as milk sours?
Casein is a kind of protein found in milk. Casein is precipitated by lowering the acidity
of the milk to a level that renders the protein insoluble while avoiding over acidification,
which results in the lactose hydrolyzing, which causes the milk to clot as it soured due to lactic
acid production.