Experiment No.

-3 (COAGULATION OF PROTEINS)
POST LAB QUESTIONS:
1. Describe and compare the appearance of the solutions.
50 ml of 10% egg white and distilled water dispersion was prepared by 5 ml egg white
and 45ml distilled water and then 5 ml was added to 6 different test tubes labelled at
different temperatures 55℃, 60℃, 63℃, 65℃, 68°C and control(unheated). Then
temperature labelled test tubes were kept in water bath set at a desired temperature for 30
min. After 30 min., it was observed that the dispersion remains unchanged in unheated
control, there is no change in color and turbidity in the control. As the temperature rises
from 55°C-68°C, the dispersion becomes more turbid and the color of solution becomes
white.

Temperature (°C) Appearance

Control (Unheated) Unchanged dispersion (clear solution)

55 Clear solution as unheated control

60 A little bit cloudy then previous solution

63 Slightly cloudier

65 Almost pure white but still cloudy

68 Pure white

2. Plot temperature Vs Absorbance and compare the curve to visual observations. You
can use either electronic spreadsheet or scale paper to plot the data points.
On plotting a Temperature Vs Absorbance graph, a sigmoidal curve was obtained from
the data provided. As the temperature increases from 55 to 60°C, there is a slight
elevation in the curve with a surge in absorbance from 0.001 to 0.181. The curve shows
altitude in absorbance as the temperature reaches to 63°C with a value of absorbance
 equal to 0.309 and follows the similar trend as the temperature reaches to 65°C with an
absorbance of 0.692. There is an extreme rise in the curve as the temperature increased to
68°C having an absorbance value equal to 1.19. This shows with increasing temperature,
protein denaturation escalates, and turbidity rises.

Temprature vs absorbance value

1.4

1.2 1.19

1
Absorbance

0.8
0.69
0.6

0.4
0.31
0.2 0.18

00
55 60 63 65 68
Temperature (℃)

3. From the data, what is the effect of heat on proteins. Explain the pattern at all data
points.
Treatment of heat to proteins results in protein denaturation and then coagulation of
proteins after cooling a heat denatured protein. After denaturation of protein due to heat
there is the change in state of protein solution sample is coagulation which form liquid
solution to solid or semi-solid solution. Different temperatures are required for different
proteins to denature varying on the nature, pH and purity of protein. Protein structure is
disrupted by the heat by breaking hydrogen bonds and additional other interactions who
maintain the protein structure. Most of the proteins can be denatured at 45-50°C and its
rate of denaturation increases at 55°C. Non-covalent bonds are broken at lower
temperatures resulting in protein unfolding and as the temperature rises disulfide bonds
begin to break. In protein denaturation 10°C increase in temperature range effects in 600-
fold surge. Albumin is the major protein in egg white which occur as 3-D shape due to
interactions formed between the amino acids present in it. Application of heat disrupts the
protein structure by breaking the hydrophobic interactions between the amino acids The
 hydrophobic amino acids will stick to one another to form a protein network trying to get
away from the water surrounding them in the egg white, that gives the egg white structure
while turning it white and opaque[ CITATION Khand \l 4105 ]. The dispersion becomes
turbid at 55°C and is clear in unheated control. Turbidity in dispersion increases as the
temperature increases resulting in opaquer, white and gel-like dispersion at 68°C with
high absorbance due to protein coagulation present in egg white.

DISCUSSION
In this experiment, the lab data came out to be different from the expected data because of some
error during the experiment. The reasons why the lab data results might be inaccurate is that the
water bath of 65℃ in which egg white dispersion was placed was no accurately calibrated which
is why the absorbance of that egg white dispersion was not precise and was extremely less than
the expected absorbance.

 
REFERENCES
FST 224 Food Chemistry lab manual (2020) ecentennial. Coagulation of proteins, Centennial
college, Scarborough, Ontario. Retrieved February 12, 2020 from
https://e.centennialcollege.ca/d2l/le/content/509251/viewContent/5412198/View

Khanacademy. (n.d.). Orders of protein structure. Retrieved February 12, 2020, from
https://www.khanacademy.org/science/biology/macromolecules/proteins-and-amino-
acids/a/orders-of-protein-structure