Acknowledgement

Any accomplishment requires the effort of many people and this project is no different
after enclosing this assignment. I am very glad and very much appreciate the support and
guidance provided to me by many people and ceaseless indebted to a great many
individuals, though space prohibited mentioning everyone who so kindly assisted in this
venture, I wish to express my appreciation to at least some of those individuals.

Writing this project was a labor of love and there are so many people I owe for helping me
and guiding me to successfully completion of this project.

I firstly would like to thank our school library and computer facilitators for free access of
information and same goes to chemistry teachers for guiding and helping in every needs.
Furthermore, I would like to thank to my friends for helping me to carry out the
experiment, and their parents for giving me the resources.

I owe huge gratitude to my guider Mr. Pema Dorji Tamang for allowing me to come up
with our own interested topic. I am very much pleased with the school management for
giving us full support as well as for giving us the opportunity to use library. And also to all
the people mentioned above for they have created miracle and lead magic happen, and for
such hard work and belief in my work.

I take this opportunity to express my sense of gratitude for my brother’s invaluable

guidance, constant encouragement, constructive comments, sympathetic attitude and
immense motivation, which has sustained my efforts at all stages of this project work.

I can’t forget to offer my sincere thanks to my family who helped me to carry out this
project work successfully and for their valuable advice and support, which I received from
time to time.

Thank you all once again…

1|Page
Table of content
Sl. No. Content Page No

1 Acknowledgement 1

2 Hypothesis 3

3 Introduction 3-5
4 Whey VS. Casein protein 5-6

5 Whey and casein are better together 6-8

6 Milk or casein allergy prevention 8

7 Uses of casein 8-9

8 Aims, materials, procedures and observations 9-13

9 Result and Conclusion 13-14

10 Bibliography 15

2|Page
 Hypothesis
If protein contained in the buffalo’s milk is more comparing to other sources, then the
quantity of casein present in it will also be more than that of cow, goat, or sheep.

Introduction
Nowadays many people are suffering from different types of diseases such as allergies, cough
and cold and many other water and air borne diseases. Those are the reasons due to lack of
diets and proteins in their body. They consume more of carbohydrates and less of other
proteins, there is imbalance in the diet.

This leads to many illness and become old at very young age. So, I came up with this research
in order to help the people by providing good information about the diets and proteins. Milk
contains proteins consisting of casein and whey. They help us in our body to become healthier
and maintain diets.Milk contains two types of proteins that is casein and whey. Casein is 80%
of the milk protein, while whey is 20%. Casein protein is digested slowly, while whey
protein digested quickly. Much like whey, casein has been repeatedly shown to increase
muscle growth and strength when combined with resistance training. It may also help with fat
loss.
Some preliminary studies have found that casein can have other impressive benefits,
including:

 Antibacterial and immune benefits: Some cell studies suggest it may provide
antibacterial and immune benefits and reduce high blood pressure.
 Triglyceride levels: One study in 10 overweight individual is found that it reduced
triglyceride levels after a meal by 22%.
 Reduction in free radicals: Some of the peptides in casein protein powder may have
antioxidant effects and fight the build-up of harmful free radicals.
 Fat loss: One 12-week training study found the average fat loss among people taking the
supplement was three times greater than in a placebo group.

Different sources of milk contain different quantity of proteins. Hence, I am going to do

research on proteins of milk that which source gives more proteins. That’s how I landed up
with this topic with the notion that it might help our poor people. Since milk is one of the
daily consumption of the people, so with the intention that it may help the society by studying
about milk and its protein and its effects to our body.

3|Page
Background Information
Milk is a white fluid secreted by the mammary glands of living organisms. It is the food of
exceptional inters probability. Milk is also contains all sorts of micro-nutrients essential for
the body of an organism. The major milk protein casein is found only in milk and nowhere in
the world. The composition of casein is not constant and depends upon the source of milk.

Average composition of milk from different sources is given below:

Source Water Mineral Protein Fats Carbohydrates
of milk % % % % %
Cow 87.1 0.7 3.4 3.9 4.9
Sheep 87.4 0.2 1.4 4.0 4.9
Goats 87.0 0.7 3.3 4.2 4.8
Buffalo 87.5 0.8 4 4.5 5.2

Casein is a major protein constituent in milk and is a mixed phosphor-protein. Casein has
isoelectric pH of about 4.7 and can be easily separated around this isoelectric pH. It readily
dissolves in dilute acids and alkalies. Casein is present in milk as calcium caseinate in the
form of micelles. These micelles have negative charge and on adding acid to milk the
negative charges are neutralized.

Ca2+-Caseinate + 2CH3COOH (aq) Casein+(CH3C00)2Ca

Casein, from Latincaseus “cheese” is the name for a family of related phosphoproteins
(ds1, ds2, B, K). These proteins are commonly found in mammalian milk, making up 80%
of the proteins in cow’s 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 amino acids,
carbohydrates, and the two inorganic elements calcium and phosphorus.

Composition
Casein contains a fairly high number of praline residues, which do not interact. There are
also no disulfide bridges. As a result, it has relatively little tertiary structure. It is relatively
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
micelles in a sense that hydrophilic parts reside at the surface and they are spherical.

4|Page
 However, in sharp contrast to surfactant micelles, the interior of casein micelles is highly
hydrated. The casein in the micelles is held together by calcium ions and hydrophobic
interactions. Any of several molecular models could 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 microvellosities 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.

The iso-electric point of casein is 4.6. Since milk’s pH is 6.6, casein has a negative charge
in milk. The purified protein is water insoluble. While it s also insoluble in neutral salt
solutions, it is readily dispersible in dilute alkalis and in salt solutions such as aqueous
sodium oxalate and sodium acetate.

The enzyme trypsin can hydrolyze a phosphate containing peptone. It is used to form a type
organic adhesive.

Whey vs. casein protein

Proteins provide the body critical amino acids that serve as building blocks for the
formation of new muscle. But not all dietary proteins are equal. The major proteins
in milk are casein and whey. These two milk proteins are both excellent sources of
all the essential amino acids, but they differ in one important aspect that whey is a
fast-digesting protein and casein is a slow-digesting protein.

5|Page
 Whey stimulates protein synthesis
Fast-digesting whey means it is emptied from the stomach quickly, resulting in a
rapid and large increase in plasma amino acids. This translates into a quick but
transient increase in protein synthesis, while protein breakdown is not affected.
Whey also has higher levels of leucine, a potent amino acid that stimulates protein
synthesis. Whey protein is superior at augmenting protein synthesis rapidly, but this
positive effect is short-lived. Consuming repeated doses of whey allows for
sustained high levels of blood amino acids and repeated bursts of protein synthesis
that provide superior effects on muscle protein balance.

Casein offers a positive protein balance

Casein is the most abundant protein in milk. It is relatively insoluble and tends to
form structures called micelles that increase solubility in water. During the
processing of milk, which usually involves heat or acid, the casein peptides and
micelle structure become disturbed or denatured to form simpler structures. As a
result, a gelatinous material is formed. This is the basis for why casein has a slower
rate of digestion, and results in a slow but steady release of amino acids into
circulation.

Whey protein resulted in a rapid increase in blood amino acids and protein
synthesis, but it was short-lived. Casein, on the other hand, resulted in a prolonged
increase in blood amino acids that resulted in a 34% reduction in protein
breakdown. The net protein balance remained more positive after intake of casein
protein over a 7-hour period. The superior long-lasting effect of casein was
attributed to a delayed gastric emptying and slower absorption rate from the
gastrointestinal tract to the blood.

Whey and casein are better together

Since whey rapidly increases protein synthesis and casein blocks protein
breakdown, a combination of both would be ideal.

A recent study compared the effects of supplementing with either a combination

whey and casein protein versus carbohydrate on several markers of muscle
anabolism during strength training.3 untrained men participated in a 10-week
resistance training program and either supplemented with 40 grams of carbohydrate
or 40 grams of protein containing a mixture of whey and casein. Half of the
supplements were consumed one hour before and then immediately after exercise
on workout days. The results were overwhelmingly positive for the combination
protein group. Despite similar background diets and identical training programs,
supplementation with protein resulted in greater increases in several measures of
muscle anabolism, including greater increases in lean muscle mass, thigh muscle
mass, muscle strength, anabolic hormones and muscle specific proteins.

6|Page
 In a similar study that lasted 14 weeks, untrained men performed resistance training
and received either 25 grams of carbohydrate or 25 grams of a combination whey
and casein protein one hour before and immediately after exercise. The
combination protein group had significantly greater increases in muscle fiber size
compared to the carbohydrate group. These studies provide strong evidence that a
combination protein consumed before and after workouts increases muscle size.

The Effects of Casein

Casein protein is the primary protein found in milk, and is essential for human
growth and development. It contains all of the essential amino acids your body
needs and cannot synthesize on its own. According to the Protein Digestibility-
Corrected Amino Acid Score system created by the World Health Organization,
casein is one of the best proteins you can consume in terms of digestibility and
nutritional value.

Nitrogen Retention
The micellar structure of casein protein is different from many other proteins in that
when you disrupt it from its suspension in milk, it will clot together. This process
occurs in cheese-making and in your intestines after you consume milk. The
clotting of casein protein enables it to break down slowly in your body, releasing
amino acids continuously into your bloodstream over several hours. Because of
slow breakdown, your body retains a high amount of the nitrogen from the casein.

Muscle Protein Synthesis

Casein protein is popular as a supplement with athletes and bodybuilders because it
is effective at increasing muscle growth when you use it in conjunction with a
resistance training program. According to Jay Hoffman, Ph.D., consuming casein
protein has been shown to increase your muscle protein synthesis rate by 31 percent
after ingestion, and these increases may last up to seven hours or more. This will
help your muscles grow at a faster rate than if you engaged in weight training
without additional protein supplementation.

Dialysis for kidney disease

One potentially harmful effect of casein protein is kidney damage. Casein is

generally safe to consume if you have normally functioning kidneys, even at levels
as high as 2.8 grams per kilogram of bodyweight according to some research. If
you already have existing kidney damage, however, the additional renal load from
casein’s nitrogen byproducts can exacerbate your condition. A 2004 study

7|Page
 examining the effects of casein protein on rats with kidney disease found that a diet
of just 20 percent casein protein caused the rats to develop renal insufficiency,
while a 20 percent soy protein diet actually improved kidney function.

Milk Allergies
For some individuals, the casein protein in milk can trigger allergies. This negative
side effect crops up when your body identifies casein protein as harmful and
releases histamine to neutralize it. The Allergy Society of South Africa notes that
both types of protein found in milk, whey and casein can trigger an allergic
reaction. Speak to your doctor if you notice allergic symptoms like hives, runny
nose, itchy eyes or rashes after consuming casein protein.

Uses of casein
1. Paint figure (casein preparation)
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 until the late 1960s when, with the advent of acrylic
paint, casein became less popular. It is still widely used by science painters,
although acrylic has made inroads in that field as well.

2. Glue
Casein-based glues, formulated from casein, water, hydrated lime and sodium
hydroxide were popular for wood working, including for aircraft. Casein glue is
also used in transformer manufacturing (especially transformer board) due to its oil
permeability. While largely replaced with synthetic resins, casein-based glues still
have a use in certain applications, such as laminating fire-proof doors and the
labeling of bottles.

3. Cheese making
Cheese contains of proteins and fats 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 proteolytic
enzyme known as rennin; traditionally obtained from the stomachs of calves, but
currently produced more often from genetically modified micro-organisms.

4. Plastics and Fibers

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

8|Page
 popular in haly during the 1930s. Recent innovations such as Mulch are offering a
more refined use of the fiber for modern fabrics.

5. Medical and Dental uses

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

Experiment

Theory
Natural milk is an opaque fluid secreted by the mammary glands of female mammals. 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 the taste. However, when
it keeps for longer time at temperature of 5 degree Celsius it becomes 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 is sufficient and temperature is around 36
degree, it forms semi-solid mass, called curd.

Aim
To study the quantity of Casein present in different samples of milk

Requirements
 Beakers (250 ml)
 Filtration Flasks
 Measuring Cylinders
 Glass rod
 Spatula
 China Dish
 Dropper
 Digital Balance
 Different samples of Milk
 Conc. Acetic Acid
 Saturated Ammonium Sulphate

9|Page
 Safety Measures
 Accidental release measures, personal precautions, protective equipment and emergency
procedures: Use personal protective equipment. Avoid dust formation. Avoid breathing
vapors, mist or gas. Ensure adequate ventilation.
 Avoid too much of inhalation of gas of concentrated ammonium sulphate
 Discharge into the environment must be avoided
 Methods and materials for containment and cleaning up: Pick up and arrange disposal
without creating dust. Keep in suitable, closed containers for disposal.

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 solution slowly and with
stirring. Fat along with Casein was precipitate out.
2. The solution was filtered and transferred the precipitates in another beaker. Add
about 30 ml of water to the precipitate. Only Casein dissolves in the water
forming milky solution leaving fat undissolved.
3. The milky solution was heated to about 40oc and adds 10ml acetic acid solution
drop-wise, when casein got precipitated.
4. Filtered the precipitate, washed with water and the precipitate was allowed to dry.
5. Weigh the dried solid mass in a previously weighed watch glass.
6. The experiment was repeated with other samples of milk.

10 | P a g e
Observation
Sample No. Source Content of Protein Percentage of Casein

(in gram)

1 Sheep’s Milk 0.55 2.7

2 Cow’s Milk 0.60 3.00

3 Goat’s Milk 0.65 3.25

4 Buffalo’s Milk 0.85 4.20

11 | P a g e
12 | P a g e
 Protein vs. Casein
4.5

3.5

2.5
Content of protein
2 Percentage of casein
1.5

0.5

0
Sheep's milk Cow's milk Goat's milk Buffalo's milk

Result
A different sample of milk contains different percentage of casein. The quantity of casein present
is directly proportional to the size of the sources.

13 | P a g e
Conclusion
As we could easily get from the observation table that the quantity of casein present differs from
different samples of milk. The buffalo has the highest percentage of casein as compared to other
samples.

So, I am very confident to say that my hypothesis was correct. If we consume more of buffalos
milk than it would help us twice better than other sources like cow, sheep etc.

Despite the difficulties I was facing, I have got many experiences particularly on the
experimental part. I could not afford to forget how much I have learnt through this project work.
I have got better views regarding the proteins that contained in the milk of mammals. It has got
many advantages than other sources.Casein is a slow-digesting protein that can boost muscle
growth and aid recovery after exercise.

Taking milk casein can improve your health, as well as increase your total daily protein intake.
This is an important factor in weight loss and muscle growth. Try taking 1–2 scoops of
casein protein powder or a large glass of milk before bed time to improve recovery and reduce
protein breakdown. At the end of the day, casein is a highly under rated source of quality protein.
You won’t be disappointed if you try it.

14 | P a g e
Bibliography
 Allen, T. (1974). Psychotherapeutic Treatment in Quality of Casein :New York
 Bergmann, P. G. (1993). The Book of Casein (Vol.26, pp.501-508). Chicago: World
Book.
 Henry, W. A., III. (1990). Sensory Isolation of Casein.135, 28-31.
 Norlander, T. (1998). Effects of Casein. New York: Scholastic Library Publishing.
 Pettingill, O. S., Jr. (1980). World Book encyclopedia. (pp. 150-155). Chicago: World
Book.

15 | P a g e