EXPERIMENT 5

DENATURATION OF EGG WHITE PROTEIN

Introduction
Egg white is of great interest for many culinary and industrial applications. Egg white is used
for coating, gluing, thickening and so on in pasta, desserts, etc. There is thus a great interest from
the industrial point of view to better know this raw material, used in very large amounts in the
dessert production for example, and to obtain egg white fractions with different functional
properties.
Protein enzymes, like militant labourers, wont work if the conditions arent right. Too hot, too
cold, or too much pressure, and their delicately tuned molecular structures turn to useless
spaghetti. You see the problem every time you cook an egg. Egg white is largely a protein called
albumin, and heat changes it from a translucent, soluble form to a solid white mass. In cooked
egg white, albumins exquisitely folded chain of amino acids is shaken apart and entangled with
others. This loss of structure in a protein molecule is called denaturation.
Objective
To study the different methods of denaturing the protein found in egg white (albumin).
Materials / Apparatus
Raw eggs
10% sodium chloride (NaCl)
10% sodium bicarbonate (NaHCO3)
1% silver nitrite (Ag NO3)
Lemon juice
Test tube
Test tube rack
Spatula
Tong
Hot plate or bunsen burner
Beaker, 400 mL
Stirring rod or glass rod
Label sticker

Procedure
1. 300 mL of water was placed in a 400 mL beaker and was heated to boil.
2. Test tubes was labeled 1 to 5.
3. 5 eggs was separated, placing the egg white in a test tube until half filled. The egg yolk was
discarded.
4. Test tube 1 was placed in the boiling water and was allowed to cook till egg turns white.
5. 5 mL of 10% sodium chloride was added to test tube 2 and stirred.
6. 5 mL 0f 10% sodium bicarbonate was added to test tube 3 and stirred.
7. 5 mL of lemon juice was added to test tube 4 and stirred.
8. 5 mL of 1% AgNO3 was added to test tube 5 and stirred.
9. The observations was recorded on the table below.
Results
EXPERIMENT 5: DENATURATION OF EGG WHITE PROTEIN
Table 5.1: Denaturation of Egg White Protein
Test Tube
1
2
3
4
5

Treatment
Heat
NaCl ionic compound
NaHCO3 base
Vinegar acid
AgNO3 heavy metal

Observations
White solid
Clear and have very little
white precipitate
Little white precipitate
A lot of white precipitate
White precipitate layer on the
surface

Discussions
Denaturing is the process of breaking down the tertiary and secondary structure of the protein
by applying some sort of external force to separate its hydrogen bonds. In the proteins found in
egg albumin, denaturing will cause it to turn from a clear liquid into a more solid white color.
For our experiment, we studied the denaturing of proteins in egg albumin. We did this by
comparing the effects of different methods proteins denaturation of egg albumin. Proteins can be
denatured by a number of different things, including heat, strong acids or bases, organic
compounds, and heavy metals. For this experiment, we cooked the egg albumin in boiling water
to test the effect of heat, vinegar to test the effect of an acid, sodium bicarbonate (NaHCO) to test
the effect of organic compounds, silver nitrate (AgNO3) to test the effect of a heavy metal, and
salt (NaCl) to test the effect of organic compounds.
With heat, the tertiary structure of the protein is manipulated because heat increases kinetic
energy, thus causing the molecules to vibrate so rapidly that the amino acid bonds are disrupted.
The application of heat resulted in a solidly gelatinous, white material; it is exactly what one
would find the egg whites to be like should he cook a hard-boiled egg. The disruption of
hydrogen-bonds and hydrophobic interactions makes the albumin in egg white denatures faster
and almost instantly turns the clear egg albumin to white solid. We then mixed NaCl in another
test tube, and rapidly shook it. After the shaking, only slight signs of denaturation were indeed
present, this is because the salt concentration is not too high and not that sufficient to disrupt salt
bridges that can make the denaturation of egg albumin more visible. When you apply an ionic
compound, the amino acids become separated from each other, thus also manipulating the
structure of the protein. Denaturation in acids and bases is viable, especially when scientists have
tested its reactant properties with salt bridges, a combination of two noncovalent interactions:
hydrogen bonding and electrostatic interactions. The application of NaHCO produced similar
results to the NaCl, but the NaHCO resulted in a slightly larger volume of denaturation. Upon the
application of an acid or base, the amino acid structure becomes straightened. In another test,
scientists found that heavy metals have the potential to disrupt disulfide bonds because of their
affinity for sulfur, thus also making it a potential catalyst for protein denaturation. That is why a
clear white precipitate layer can be clearly seen on top of the surface of the egg white.
Conclusion
In conclusion, this experiment proved that all methods used would cause a denaturing effect in
the albumin, but the degree of the denaturation is different. From our observation, the method to
denature the protein in egg white can be clearly seen by using heat, and then by using silver
nitrate (AgNO3). Methods of adding NaCl and NaHCO3 only cause a little denaturation in egg
albumin. The denaturation of egg albumin by using vinegar shows a slightly greater denaturation
effect that NaCl and NaHCO3 because the vinegar have high concentration of acid.

Questions
1. Which method appeared to have the most dramatic affect on egg albumin? Why do you think
this method had a greater affect?
The source that produced the most effective denaturation results was the heat. A possible reason
for this is that the heat produces much kinetic energy, which in turn gives you a fast reaction
because of the extents to which the amino acids are affected. On the other hand, the ionic
compounds, bases, etc. would produce a slower result because of the fact that kinetic energy is
not present to further facilitate the process.
2. Of the methods you tested, which would be more likely to be used in the food industry?
In our food industry, it is most likely to use the method of heat, as in fact, boiling an egg is
already a favorite of many when it comes to different cooking methods. The application of heat
is safe for the food industry to use because the use of any external elements (elements referring
to chemicals such as AgNO3) could cause potential health complications in consumers.
References
- Protein denaturation, Proteins unravelled, December 2009, Chemistry World.
- www.chemistryworld.org
- M. Ferreira, C. Hofer and A. Raemy, A Calotimetric Study Of Egg White Proteins, Escola
Superior de Biotecnologia, R. Dr. Ant6nio Bernardino de Almeida, 4200 Porto, Portugal, Nestle
Research Center, Vers-chez-les-Blanc, EO. Box 44, CH-100 Lausanne 26, Switzerland.
- Zoubida Akkouche, Lyes Aissat, Khodir Madani, Effect of Heat on Egg White Proteins, Faculty
of Nature and Life Sciences, 3BS Laboratory A. Mira University, Bejaia, Algeria.