Isolation of Casein and Albumin
Isolation of Casein and Albumin
Isolation of Casein and Albumin
ABSTRACT
Selected proteins were isolated from their commercially available sources using different isolation techniques for each.
The group was assigned to isolate the proteins casein and albumin from a sample of skimmed or non-fat milk by
means of isoelectric precipitation and heat denaturation, respectively. The group used a 50 mL sample of Nestle nonfat milk which was first heated to 40 oC, after which 10% acetic acid was added dropwise until a curd-like precipitate
was formed, indicating that the isoelectric pH of casein has been reached. The precipitated casein was filtered by
gravity filtration and appeared as a white amorphous precipitate. The filtrate from which albumin was isolated
appeared as a cloudy yellowish liquid. It was placed in a 75 oC water bath. No precipitate was formed as the liquid
stayed in the water bath longer than the indicated 5 minutes.
INTRODUCTION
Milk is one of the substances that are abundant
in different types proteins. The primary protein
component of milk is a protein group called
caseins. Casein is the general term for a family of
related phosphoproteins. These make up about
80% of the proteins in cows milk and around 2045% of the proteins in human milk. The casein
family of proteins consist of several types of
casein, namely -s1, -s2, , and 6 [3]. These
form a multi-molecular granular structure called a
micelle, along with water and some salts [1].
They bear a specific amino acid composition that
is essential to for growth and development, which
makes milk one of the most essential
components of a healthy human diet. One of the
significant properties of caseins is that it
precipitates or coagulates at pH 4.6.
Aside from this, certain albumins can also be
found in milk in the form of lactalbumins.
Albumins are globular proteins that are soluble in
water and in dilute salt solutions and have the
ability to become denatured and coagulated by
heat application [2]. A specific example of a
lactalbumin is -lactalbumin, which is classified
as a whey protein. -Lactalbumin plays an
important role in the synthesis of lactose and is
essential in the process of milk synthesis [1].
Once casein has been isolated from the milk and
the milk has been made acidic, lactalbumin can
be precipitated out by heating the mixture.
In this experiment, several proteins were
isolated from their commercially available sources
which is needed in order to be able to study their
structure, properties, and different functions of
proteins. There are various methods by which
proteins can be isolated that involve different
principles. One of these methods is isoelectric
precipitation. The isoelectric pH (pI) is the pH
value at which the protein has zero net charge.
Isoelectric precipitation is done by adjusting the
pH of the protein source until the pI is reached.
This reduces the solubility of the protein, since it
EXPERIMENTAL
A. Sample used
Nestle Non-Fat Milk
B. Procedure
1. Isolation of casein from skimmed
milk
50 mL of the liquid milk sample was placed in a
100-mL beaker, and then heated on a hot plate
while maintaining the temperature at 40oC. Upon
reaching this temperature, 10% acetic acid was
added dropwise into the milk sample while the
solution was stirred gently after every 5 drops.
This was done continuously until the pH of the
2.
after
water
DESCRIPTION
Clumps of yellowish white solid
precipitate (amorphous)
Yellow cloudy liquid
of
the
Figure
Albumin filtrated
heating in
bath
The principle
behind the
separation
albumin involves
REFERENCES
[1] Hurley, Walter L. Milk Composition: Proteins.
http://ansci.illinois.edu/static/ansc438/Milkco
mpsynth/milkcomp_protein.html 03/28/2016
[2] Minard, R. Isolation of Casein, Lactose and
Albumin
from
Milk.
http://courses.chem.psu.edu/chem36/New
%20Syn%2036%20pdf/Exp112.pdf
03/29/2016
[3] n.a. Milk Protein.
http://www.milkfacts.info/Milk
%20Composition/Protein.htm 03/29/2016
[4] Ophardt, Charles E. Denaturation of Proteins.
http://chemistry.elmhurst.edu/vchembook/56
8denaturation.html 03/28/2016
[5] Young, Derek R. Isoelectric Precipitation.
http://www.rpi.edu/dept/chem-eng/BiotechEnviron/PRECIP/precpph.html 03/28/2016
[6] Young, Derek R. Salt Induced Precipitation of
Proteins.
http://www.rpi.edu/dept/chemeng/Biotech-Environ/PRECIP/precpsalt.html
03/29/2016