INDEX

Sr.NO. TOPIC PAGE No.

1. INTRODUCTON 2-3
2. COMPOSITION 4-5
3. Structure 6-7
4. Models 8
5. Characteristics 9
6. USES 10-14
7. CONTROVERSIES 15-18
8. EXPERIMENT 19-22
9. BIBLIOGRAPHY 23

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Casein is the name of related phosphor proteins. These
proteins are commonly found in mammalian milk,
making up 80%of the proteins in cow milk and between
20% and 45% of the proteins in human milk. Casein has
a wide variety of uses, from being a major component
of cheese, to use as a food additive, to a binder for
safety matches. As a food source, casein supplies for
amino acids, carbohydrates and two inorganic
elements, calcium and phosphorus.

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 Casein is a complete protein meaning that it contains
all of the essential amino acids, which the body cannot
manufacture on its own. When dried, it is a white,
amorphous powder without taste and odour. It is a
mixed phosphoprotein and occurs in milk as calcium
salt (calcium caseinate) in the form of micelle. The
micelle has a negative charge. When an acid is added to
the milk, the negative charges are neutralized.

Calcium caseinate + acetic acid → casein (s) + calcium acetate(aq)

The quantity, quality and fat-content from the

various milk samples differ with the type of particular
mammals and their fodder.

Milk casein in powdered form

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 Casein contains a fairly high number of proline
residues, which do not interact. There are also no
disulfide bridges. As a result hydrophobic, making it
poorly soluble in water. It is found in milk as a
suspension of particles called “casein micelles” which
show only limited resemblance with surfactant-type
micellae in a sense that the hydrophilic parts reside at
the surface and they are spherical. However, in sharp
contrast to surfactant micelles, the interior of a casein
micelle is highly hydrated.

Casein micelles under electron microscope

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The caseins in the micelles are held together by calcium
ions and hydrophobic interactions. Several models
account for the special conformation of casein in the
micelles. One of them proposes the micellar nucleus is
formed by several sub micelles, the periphery
consisting of microvellosites of K-casein. Another
model suggests the nucleus is formed by casein-
interlinked fibrils.

Finally, the most recent model proposes a double link

among the caseins for gelling to take place. All three
models consider micelles as colloidal particles formed
by casein aggregates wrapped up in soluble K-casein
molecules.

Casein micelle is composed of four types of protein,

namely alpha s1, alpha s2, beta and kappa casein.

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The four casein molecules (αS1-, αS2-, β-, and κ-caseins)
are well known in terms of amino acid composition,
molecular weight, concentrations in milk, and global
physico-chemical properties. All these casein molecule
are more or less phosphorylated. This confers to these
proteins a good capacity to bind calcium phosphate.
Another important characteristic is the glycosylation of
5096 of K—casein. Due to this glycosylation, its C-
terminal part is hydrophilic. This characteristic is
important for the micellar structure and stability. On
the other hand, casein molecules have high contents in
prolyl residues conferring open and flexible
conformations. Due to their high flexibility, casein
molecules have excellent surface-active and stabilizing
properties.

Structure of casein

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 The casein molecules are associated together to form
casein micelles. The casein micelles contain αs1-, αs2-,
β-, and κ-caseins in proportions 40, 1o, 35, and 15%
(w/w), respectively. Casein molecules are associated
together by protein-protein interactions (hydrophobic,
hydrogen and electrostatic binding) and also by the
presence of calcium and phosphate (6–8% in weight).
They have an average diameter close to 150 nm with a
huge polydispersity, a zeta potential of about −20 mV,
and a high amount of associated water
(4 g of water/g of protein).

Casein micelle and submicelle

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 Different models of micellar organization were
proposed thanks to the improvement of the
methods used to their characterization. The main
proposed models are:

(1) A submicellar model where the micelles correspond

to submicelle assemblages linked together by
nanoclusters of calcium phosphate (Morr; 1967a;
Schmidt 1982; Ono and Obata 1989; Walstra 1999).

(2) A model with an open structure (Holt and Horne

1996).

(3) An open model like sponge (Marchin et a1. 2007;

Bouchoux et a1. 2010; Dalgleish 2011).

In this last proposed structure, dense regions

composed of caseins in interaction with nanoclusters of
calcium phosphate and water channels containing salts
and caseins are described. In all of these models, the
glycosylated forms of K-casein are located at the
surface of casein micelles, conferring them a negative
charge and stability due to electrostatic repulsion and
steric hindrance.

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Main physico-chemical characteristics of casein micelle:

Sr. No. Characteristics Values

1. Diameter 50-150 nm

2. Surface area 8 × 10−10 cm2

3. Volume 2.1 x 10-15 cm

4. Density(hydrated) 1.0632 g.cm-3

5. Mass 2.2 x 10-15 g

6. Water content 63%

7. Hydration 3.7 g H2O.g-1

Molecular
8. 1.3 x 109 g.mol-1
weight(hydrated)
Molecular
9. 5 x 108 g.mol-1
weight(dehydrated)
10. Mean free distance 240 nm

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 1. PAINT
Casein paint is a fast drying, water soluble medium
used by artists. Casein paint has been used since
ancient Egyptian times as a form of tempera paint, and
was widely used by commercial illustrators as the
material of choice until the late 1960s when, with the
advent of acrylic paint, casein became less popular. It is
still widely used by scene painters, although acrylic has
made inroads in that field as well. Casein paste paints,
to be diluted with water, are made by pigmenting
weakly alkaline solutions of casein. Casein is also used
as the emulsifying agent in emulsion paints, which are
oil paints that can be thinned with water.

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 2. GLUE
Casein- based glues were popular for woodworking,
including for aircraft, as late as the de Hacilland
Mosquito. Casein glue is also used in transformer
manufacturing (especially transformer board) due to its
oil permeability. While largely replaced by synthetic
resins, casein-based glues still have a use in certain
niche applications, such as laminating fireproof doors
and the labeling of bottles.
Casein paste paints, to be diluted with water, are made
by pigmenting weakly alkaline solutions of casein.
Casein is also used as the emulsifying agent in emulsion
paints, which are oil paints that can be thinned with
water.

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 3. CHEESE MAKING
Cheese consists of proteins and fat from milk, usually
the milk of cows, buffalo, goats or sheep. It is produced
by coagulation of casein. Typically, the milk is acidified
and then coagulated by the addition of rennet,
containing a protrolytic enzyme, typically obtained
from the stomachs of calves. The solids are separated
and pressed into final form. Unlike many proteins,
casein is not coagulated by heat. During the process of
clotting, milk-clotting proteases act on the soluble
portion of the caseins, K-casein, thus originating an
unstable micellar state that results in clot formation.
When coagulated with chymosin EC 3.4.23.4) is an
aspartic protease that specially hydrolyzes the peptide
cond in Phe105-Met106 of K-casein, and is considered
to be the most efficient protease for the cheese-
making industry (Rao et al., 1968).

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 4. PLASTIC AND FIBER
Some of the earliest plastics were based on casein. In
particular, galalith was well known for use in buttons.
Fiber can be made from extruded casein. Lanital, a
fabric made from casein fiber (known as Aralac in the
United States), was particularly popular in Italy during
the 1930s. Recent innovations such as QMilch are
offering a more refined use of the fiber for modern
fabrics. Molded plastics of casein resemble horn, ivory,
ebony, and marble in appearance. The principal use is
for buttons for clothing. Wool-like textile fibers are
made from casein by extruding an alkaline solution of
casein through spinnerets of the kind used in making
rayon and then hardening the fibers in an acid bath
containing formaldehyde. There was renewed interest
in casein fiber in the 21st century as manufacturers
sought to explore sustainable alternatives to
petroleum-based synthetic fibers such as nylon.

White GALALITH Australian

Royal Airforce pre-1953

Buttons

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 5. PROTIEN SUPPLEMENTS
An attractive property of the casein molecule is its
ability to form a gel or clot in the stomach, which
makes it very efficient in nutrient supply. The clot is
able to provide a sustained slow release of amino acids
into the blood stream, sometimes lasting for several
hours. Often casein is available as hydrolyzed casein,
whereby it is hydrolyzed by a protease such as trypsin.
Hydrolyzed forms are noted to taste bitter and such
supplements are often refused by infants and lab
animals in favor of intact casein.

6. MEDICAL AND DENTAL USES

Casein-derived compounds are used in tooth
remineralization products to stabilize amorphous
calcium phosphate (ACP) and release the ACP onto
tooth surfaces, where it can facilitate remineralization.

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 CONTROVERSIES AND SIDE-
EFFECTS

1. AUTISM
Although research has shown high rates of use of
complementary and alternative therapies for children
with autism, including gluten and/or casein exclusion
diets, as of 2008 there is a lack of evidence for the
efficacy of these diets. A 2006 review of seven studies
indicated that, although all reported benefits of
exclusive diets in reducing autism symptoms, all
suffered design flaws, and there was not enough
evidence overall to justify recommending exclusion
diets to patients.

3. CANCER
T. Colin Campbell’s The China Study (2005), a book,
describes a direct correlation between casein
administered to rats and the promotion of cancer cell
growth when exposed to carcinogens. Aflatoxin

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(a potent carcinogen) was administered to these rats
over a 2 week dosing period. The rats were given a 1
week post-dosing period before beginning the test
(promotion period). During the promotion period, one
group of rats was put on a 5% casein protein diet and
another group on a 20% casein protein diet. None of
the rats on 5% casein protein developed foci,
precursors to cancerous cell growth, and every rat on
20% casein protein developed the precancer foci. It
should be noted that all test groups were fed a 20%
casein diet for a total of 5 weeks (2-wk acclimation, 2-
wk dosing, 1-wk post-dosing) prior to the 12 week
promotion period in order to survive the initial
aflatoxin B1(AFB1) dosing, regardless of whether they
were in the 5% or 20% test groups. Campbell has
performed additional studies using a range of different
carcinogens and other experimental animals, and
claims to have found a consistent correlation between
cancer growth and the amount of casein protein in
diet. A 2001 study suggests another milk protein, when
protein, may play a protective role against colon
tumors in rats. According to a study from the
Australian Dairy Council, casein has antimutagenic
effects

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 3. A1/A2 beta caseins in milk
A1 and A2 beta-casein are genetic variants of the beta-
casein milk protein that differ by one amino acids; a
proline occurs at position 67 in the chain of amino
acids that make up the A2 beta-casein, while in A1
beta-casein a histidine occurs at that position. Due to
the way that beta-casein interacts with enzymes found
in the digestive system, A1 and A2 are processed
differently by digestive enzymes, and a seven-amino
peptides, beta-casomorphine-7, (BCM-7) can be
released by digestion of A1-beta-casein. The A1 beta-
casein type is the most common type found in cow's
milk in Europe (excluding France), the United States,
Australia, and New Zealand.
Interest in the distinction between A1 and A2 beta-
casein proteins began in the early 1990s via
epidemiological research and animal studies initially
conducted by scientists in New Zealand, which found
correlations between the prevalence of milk with A1
beta-casein proteins and various chronic diseases. The
research generated interest in the media, among some
in the scientific community, and entrepreneurs. A
company, A2 Corporation, was founded in New

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 Zealand in the early 2000s to commercialize the test
and market "A2 Milk" as a premium milk that is
healthier due to the lack of peptides from A1. A2 Milk
even petitioned the Food Standards Australia New
Zealand regulatory authority to require a health
warning on ordinary milk.
Responding to public interest, the marketing of A2
milk, and the scientific evidence that had been
published, the European Food Safety Authority (EFSA)
reviewed the scientific literature and published a
review in 2009 that found no relationship between
chronic diseases and drinking milk with the A1 protein.
An independent review published in 2005 also found
no relationship between drinking A1 or A2 milk and
chronic diseases.

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 AIM:

To determine amount Of CASEIN

present IN DIFFERENT SAMPLES OF MILK.

REQUIREMENTS:

APPARATUS
• 250ML BEAKERS
• FUNNEL, GLASS ROD
• PORCELAIN DISH
• CHEMICAL BALANCES
• TEST TUBES
• FILTRATION FLASK
• BURNER

CHEMICALS
• DIFFERENT SAMPLES OF MILK
• 1% OF ACETIC ACID SOLUTION
• SATURATED AMMONIUM SULPHATE SOLUTION

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 THEORY:
Natural milk is an opaque white fluid secreted by the
mammary glands of female mammal. The main
constituents of natural milk are protein, carbohydrate,
mineral vitamins, fats and water and are a complete
balanced diet. Fresh milk is sweetish in taste. However,
when it is kept for long time at a temperature of 5
degree it become sour because of bacteria present in
air. These bacteria convert lactose of milk into lactic
acid which is sour in taste. In acidic condition casein of
milk starts separating out as a precipitate. When the
acidity in milk is sufficient and temperature is around
36 degree, it forms semi-solid mass, called curd. The
Protein, Casein, Exist in milk as the calcium salt,
calcium casein etc. The salt has a complex structure
and form micelles. These micelles have negative
charges on the outer surface.

Isolation of casein
from milk

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 PROCEDURE:
1. A clean dry beaker has been taken, followed by
putting 20 ml of cow’s milk into it and adding 20 ml of
saturated ammonium sulphate slowly and with stirring.
Fat along with casein was precipitate out.
2. The solution was filtered and transferred the
precipitates in another beaker. Added about 30 ml of
water to the precipitate. Only casein dissolves in water
forming milky solution leaving fat undissolved.
3. The milky solution was heated to about 40° C and 1%
acetic acid solution drop-wise, when casein got
precipitated.
4. Filtered the precipitate, washed with water and the
precipitate was allowed to dry.
5. Weighed the dry solid mass in a previously weighed
watch glass.
6. The experiment was repeated with other samples of
milk.

Different samples of milk

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 OBSERVATIONS:

Weight of % of
Source of Volume of
Sr.No. casein casein
milk milk taken
present present
1. Cow 20 ml 0.62 g 3.1%
2. Goat 20 ml 0.56 g 2.8%
3. Buffalo 20 ml 0.66 g 3.3%
4. Sheep 20 ml 0.9 g 4.9%
5. Camel 20 ml 0.923 g 2.7%

RESULTS:
This study clearly indicated that the amount of casein
precipitated from the sheep milk was higher than that of the
other milk samples. The quantitative analysis of casein
precipitated from the various milk samples provide the ample
scope to the cottage cheese manufacture. Thus, the milk is
suitable for the best muscle growth and basic body building
achievements.

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 WEBSITES:
 wikipedia.org
 academia.edu
 iosrjournals.org
 seminarsonly.com

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