LABORATORY REPORT

FST 261 INTRODUCTION TO FOOD CHEMISTRY

EXPERIMENT 5: DENATURATION OF EGG WHITE PROTEIN

INTRODUCTION

Proteins, which are polymers of amino acids, are crucial to the proper functioning of every
living organism. The amino acid chains that make up proteins are very lengthy and very
complex. Proteins have three different types of structure: primary, secondary, and tertiary.
The amino acid sequence is the fundamental building block of proteins. The folded state of a
protein is determined by its secondary and tertiary structures. Proteins are essential for many
cellular processes, including facilitating the transport of molecules, responding to signals,
speeding up events, and replicating DNA, but they are rendered ineffective if they lose their
appropriate folded structure (Moeller, 2018). Proteins deform easily if the right circumstances
are not met. Proteins are held in their natural structures for investigation. The protein
structure is held together by this synergy of forces. Hydrophilic interactions, hydrogen
bonds, disulfide bridges, and ionic interactions are examples of these types of forces.
Denaturation is the process of altering the structure of a protein in order to temporarily or
permanently impair its structural forces. Proteins lose their structure when subjected to
circumstances that are outside of their tolerance range. Proteins are often water-soluble in
their natural shape. This causes changes in solubility by disrupting secondary and tertiary
structures. The solution becomes cloudy and changes color to white as precipitates develop.
Coagulation is the process by which a denatured protein becomes precipitated. We can
denature the proteins by disrupting the H-bonds that are within the structure. When this
happens the overall shape of the protein changes and new properties can be observed. The
shape of a protein is associated with food processing properties, such as solubility, gel
formation, and enzyme activity. In the egg whites, the albumin will change from clear to
white. We will explore how the following denatures egg albumin.

The objectives of this experiment are to experiment with different methods of denaturing the
protein found in egg white (albumin).

Firstly, we will denature the egg white by heat which is commonly done by cooking.
Secondly, using acids and bases can form ions on some side groups of amino acids. Thirdly,
organic compounds will form their own hydrogen bonds with the amino acids. Lastly, adding
heavy metals such as silver nitrate will react with disulfide bonds and cause denaturation.

In conclusion, it is hypothesized that as more changes in color and appearance of the egg
white such as coagulation, the more denatured the egg white protein is.

MATERIALS / APPARATUS

Raw eggs 10% sodium chloride (NaCl)

10% sodium bicarbonate (NaHCO3) 1% silver nitrate (AgNO3)
Lemon juice Test tube
Test tube rack Spatula
Tong Water bath
Beaker, 400mL Vortex mixer
Label sticker

PROCEDURE

1. 300mL of water was placed in a 400mL beaker and was heated to a boil.

2. Test tubes were labeled 1 to 5.

3. 5 eggs were separated, the egg white was placed in a test tube until half filled. The egg
yolk was discarded.

4. Test tube 1 was placed in the boiling water and allowed to cook till the egg turned white.

5. 5mL of 10% sodium chloride was added to test tube 2 and stirred.
6. 5mL of 10% sodium bicarbonate was added to test tube 3 and stirred.

7. 5mL of lemon juice was added to test tube 4 and stirred.

8. 5mL of 1% AgNO3 was added to test tube 5 and stirred.

9. Observations were recorded in the table below.

RESULT

Table 5.1: Denaturation of Egg White Protein

Test Treatment Observations

tube
1 Heat The egg white turns from colorless to white color and it
becomes solid

2 NaCl – ionic Clear color remained

compound

3 NaHCO3 – base Clear color remained

4 Lemon juice – acid Formed white precipitate

5 AgNO3 – heavy metal Cloudy on top of the layer solution

DISCUSSION

Based on this experiment, the albumen in the white egg which is the protein are denatured by
5 different types of methods. Firstly, the white egg (test tube 1) was heated, followed by NaCl
ionic compound (test tube 2). Next, add NaHCO3 solution (test tube 3) and lemon juice (test
tube 4). Lastly, by adding AgNO3 to the white egg. The denaturation of protein (albumen) by
the disruption of secondary and tertiary structures in the white egg.

Based on Table 1.1, the observations by heating the white egg in boiling water, the white egg
was denatured and turned into a solid and the clear color turned to white color. The albumen
becomes denatured due to the heat breaking apart the hydrogens bond that holds the proteins
in their original shape. In the tertiary structure, the side chains such as salt bridges, non-polar
hydrophobic, disulfide bonds, and hydrogen bonds are disrupted thus breaking the helical
structure. After that, the protein is unfolded causing more space are taking up and hardening
them in place next to one another. (Moeller, Breaking Proteins, 2018)

Next, NaCl solution was added to the white egg. After the solution was added, the clear color
of the white egg remained. By adding NaCl, due to the high concentration of salt and the
presence of water, the hydrogen bonds in the white egg were disrupted by the interaction
between ions and the protein molecules thus denaturing the protein happened. This can cause
molecules of protein in the white egg to unfold and changes their original shape to become
more liquid. (Li, 2018)

After that, the third method is by adding the NaHCO3 solution, and the clear color of the
white egg has remained. This is because adding NaHCO3 increases the solubility of the white
egg. The hydrophobic residues are exposed and induce calcium binding inactivation and
protein unfolding resulting in the myofibrillar protein denatured. This also caused the loss of
the original shape and become more liquid. (Yan-Ping Li, 2021)

For Test tube 4, lemon juice was added to the white egg. The observation of this method is
the formation of a white precipitate at the top of the solution. The acidity or the pH from
lemon juice causes some of the hydrogen bonds broken and the salt bridge formed between
the side chains that hold the shape of molecules of protein is disrupted and changes the
original shape thus becoming tangled. The lemon juice also causes the protein molecules to
unfold and loses the tertiary structure. Because of this, the white egg becomes solidified and
opaque. (How do acids and bases denature a protein?, 2020) . But based on our result, the
white egg does not turn completely solid because it is not being heated.

For the last test tube (Test tube 5), after the addition of AgNO3, a cloudy precipitate was
formed at the top layer. This is due to the formation of silver albuminate (after the
combination of albumen and positive metal ion) being insoluble and causing denaturation in
the white egg. The ionic compound disrupts the salt bridges and changes the original shape of
the protein’s structure and becomes opaque and solidifies. (Kanazariya, 2018)
CONCLUSION

The objective of this experiment has been achieved successfully. This is because we did
different methods to denature the protein found in the white egg which is heating, using
sodium chloride, sodium bicarbonate, lemon juice, and silver nitrate. We also got different
results for each method we did. When heated, we observed that the white egg turned into a
white solid. Clear color remained when used sodium chloride and sodium bicarbonate. A
white precipitate was formed when using lemon juice as a treatment. Lastly, cloudy on top of
the layer solution formed when using silver nitrate.
QUESTIONS

1. Which method appeared to have the most dramatic denaturing affect on egg albumin? Why
do you think this method had a greater affect?

The heating method was the most dramatic denaturing effect on egg albumin because the
physical appearance of egg white dramatically changed in colour from transparent to white
and liquid to solid texture because the egg white has completely denatured. The increased
effectiveness can be attributed to the fact that heat can be employed to break hydrogen bonds
and non-polar hydrophobic interactions (Moeller, 2018). Heat may boost and give the kinetic
energy needed to cause the atoms in protein molecules to vibrate so swiftly and violently that
the bonds are broken. This is the quickest and most efficient way that heat may denature egg
albumin.

2. Of the methods you tested, which would be more likely to be used in the food industry?

Heating methods would be more likely to be used in the food industry because heat treatment
improved the solubility, foaming properties, and emulsifying activity of the egg white. For
example, Besides that, the heating method can completely denature egg albumin causing
bacteria that can cause food poisoning such as Salmonella can be eliminated. Hence, egg
white is safe to eat for consumers.
References
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https://www.aatbio.com/resources/faq-frequently-asked-questions/How-do-acids-and-
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Kanazariya, S. (2018, November 12). Precipitation Reactions of Protein. Retrieved from

https://www.zmchdahod.org/pdf/college/Reactions_of_Protein-01-11-2018.pdf

Li, J. Z. (2018, January 31). Combination effects of NaOH and NaCl on the rheology and gel
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Yan-Ping Li, X.-H. Z.-L. (2021). Effect of sodium bicarbonate and sodium chloride on aggregation and
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