Group Name: Date: June, 2023

Course & Year: BSF-2 Subject: Biochemistry

Laboratory Report No. 4

Denaturation of Proteins

Abstract

Proteins function in the biological system as hormones, oxygen carriers, and

catalysts or enzymes. Protein structural changes, both large and tiny, can occur
under conditions different from those of the protein's environment. Denaturation, a
process that causes structural disturbance, involves the dissociation of peptide
bonds. An overview of the various ways that protein disruption occurs is given in this
abstract. The findings of the albumin solution experimented with a water bath, NaCl
solution, NaHCO3, lemon juice, rubbing alcohol, AgNO3, detergent solution, and a
vigorous shaking will be covered in this paper.

This study also includes observations made while testing several albumin
solutions in test tubes. The test tubes with the labels 2 through 9 are tested under
various settings, with the first test tube serving as the control. The albumin solution's
color, consistency, pH level, and protein composition were all observed, as well.

The most prevalent circulating protein in plasma is albumin. In healthy human

patients, it makes up half of the plasma's total protein content. Hepatocytes in the
liver produce albumin, which is then swiftly discharged into the bloodstream at a rate
of between 10 and 15 grams per day.

An egg white that has been diluted in water is used to create the albumin
solution required in this exercise. As a result, equally distribute the 2 ml of albumin
solution among the nine test tubes. This article discusses the method, findings, and
solutions to a number of protein denaturation-related queries.
Introduction

This section offers a summary of the activity. The objective of the lab
experiment is to compare the effects of various circumstances on a 2 ml albumin
solution across nine test tubes. The task's objective is to ascertain how protein is
disrupted in diverse conditions in order to ascertain what physical or chemical agent
denaturingly affects egg albumin and what protein structure is disturbed.

Process and Procedure

The group has made an albumin solution from two eggs in this activity by
separating the egg white from the yolk. After that, cheesecloth is used to filter and
squeeze the egg white. For the albumin solution to be utilized for the subsequent
stages, a measurement of 5 ml of albumin diluted to 100 ml of water was made. The
following are the various conditions tested in 2 ml of albumin solution in each test
tube:

1. The first test tube is the control and all other test tube will be compared to it.
2. On the second test tube, place on water bath (beaker filled with water). Heat
until the water bath boils.
3. On test tube #3, add 2 ml NaCl solution to test tube #3 and shake slowly.
Record observations.
4. Add 2 ml NaHCO, to test tube #4 and shake slowly.
5. Check the pH of the egg white. Add 2 ml of lemon juice or HCI solution to test
tube #5 and shake slowly. Check the pH again..
6. Add 2 ml rubbing alcohol to test tube #6 and shake slowly.
7. Add 10 drops of 1% AgNO, to test tube #7 and shake.
8. On test tube #8, add 2 ml detergent solution and shake.
9. Stir test tube #9 with stirring rod and shake vigorously.
Results and Observations

Observations are carefully recorded when the albumin solutions are tested under
various circumstances. The findings from test tubes 1 through 9 are listed below:

Denaturation of Proteins
Name (Last name, First
name):
Group Name:
Test Tube No. Denaturing Agent Observations
1 (Control)
2 Heat The albumin coagulate, it
scattered and hardened
inside the test tube, it is
very visible and widely
spread. The color
transformed from
transparent to opaque
white.
3 KCl solution There is a minimal
compressed coagulation
of the albumin. The
solution becomes
transparent with visible
bubbles at the top.
4 NaOH solution There is a minimal spread
coagulation of the
albumin. The solution
becomes transparent with
heavy bubbles at top.
5 Lemon juice Albumin solution’s pH level
is 8.5 (Basic), after adding
the lemon juice it becomes
6.0 (Acidic). The color
changed into a pale yellow
with small bubbles at top
of the solution.
6 Alcohol The albumin coagulated
and becomes insoluble,
thus very visible to the
naked eye. The solution’s
color becomes transparent
with no bubbles.
7 AgNO3 The solution’s color
becomes creamy white
with visible bubbles. It is
 hard to determine if the
albumin coagulated or not.
8 Detergent The albumin coagulated
and becomes insoluble,
thus very visible to the
naked eye (more visible
than alcohol solution). The
solution’s color becomes
opaque white with
bubbles.
9 Mechanical stress The solution’s color
(stirring) becomes opaque white
visible bubbles and there
is a minimal spread
coagulation of the
albumin.

Following careful investigation, it was discovered that the coagulation,

hardening, and insoluble character of albumin is a sign of a denatured protein in an
albumin solution. After treatments in the solutions, the albumin's solubility drastically
decreased.

Below are the answered questions after performing the laboratory activity:

1. Which is physical/chemical agent appeared to have the most evident

denaturing effect on egg albumin? Why?
- The most pronounced denaturing action is heat on egg albumin, it is seen
in the solution, where the albumin coagulates and abruptly hardens.
Compared to other treatments, the coagulation is quite evident to the
naked eye.
2. What structure of protein is disrupted using the methods above?
- Denaturation implies the destruction of the tertiary structure of a protein
molecule and the formation of random polypeptide chains.
3. Among the methods you tested, which would be more likely to be used in the
food industry?
- Applying heat would be more likely to be used in food industry as sensory
properties are better retained by the use of higher temperatures.
4. Why heat and alcohol used to disinfect medical equipment?
 - These two possess the ability to kill microorganisms. Also, they cause cell
lysis and membrane damage in bacterias by denaturing proteins.
5. Why is milk given to someone who accidentally ingests a heavy metal ions
such as silver and mercury?
- Casein and whey, two proteins found in milk, have the ability to bind heavy
metal ions, preventing them from attacking other bodily parts. In order to
make the heavy metal ions less reactive and enable safe removal from the
body, they can also be reduced.

Conclusion

The experiment's findings demonstrated that a protein's insoluble state is a

sign of denatured protein. Heat is found to have a more pronounced denaturing
effect on an albumin egg than the other 9 treatments. This explains why medical
equipment is cleaned with heat and alcohol.

Consequently, applying treatments and waiting for the protein to congeal or harden
are two ways to denature it.
Appendices

Results

Observations
GROUP NAME:

Group members: