Food Science

Protein Denaturation and Coagulation Lab Experiment

March 19, 2015

Group Members:
June, Thanatcha Jantanapongpun
Preme, Ravisara Karnjanolarn
Mind, Poonnapha Kittiratanaviwat
Kam, Pitiporn Prachayamongkol
Gam, Nichanan Suteeduangsamorn

Introduction
Protein are necessary for our bodies by working on many function such as building up
muscles, bones, skins, hair, and also be involved in every tissues of bodies. Protein are made out

of amino acids group which are Amino group, R group and Carboxyl group. These amino acids
are linked together by the Peptide bonds that was formed by a dehydration reaction. Additionally,
Hydrogen bonds is responsible for the folding of each protein polypeptide chain together. Protein
become denatured commonly by heat or chemical reaction, they became unfolded/spread out and
coagulated into random shapes which they cannot go back to its original shape. There are many
factors that affects protein denaturation such as pressure, freezing, pH, metal ions, and
mechanical treatment. If the coagulation cannot be reversed, the process continues and changes
their state from liquids into a soft semi-solid clot or solid mass. A soft semi-solid clot or clot is
defined as mass that stuck together, and become thicken. In this experiment, they helped in
determining the changes by heat and chemical solution. We did three experiment which are
finding milk curds and Soy milk curds, and comparison of Meat texture. Between Milk and Soy
milk Curds, we were using different types of chemical substances which are Vinegar represented
acid and Magnesium Sulfate represented salt respectively. We added the solution into the beaker
of milk and soy milk, then stirred it while they were being heated. Milk and soy milk was firstly
denatured due to the increasing rates of temperature at the same time when proteins unfolded
itself. Milk has curds calls Casein - one type of proteins in milk and Soy milk has curds calls Soy
protein. Specifically, we have learned to separate the proteins from the milk by aid of chemical
solution, we need to use a particular substance that capable for that protein. For the example,
Whey protein and Casein need a different solution. Since we use only Vinegar in Milks
experiment, Casein is separated but Whey protein arent. Whey is still a liquid mixed with other
substances.
Proteins are contained especially in muscle for both human and animals, so chicken meat
have to contain its too. In this experiment, we used chicken to identify and contrast the
differences between the texture of the raw and the cooked meat which it was being boiled at high
temperature to show that the texture and shape of meat was denatured.
To conclude, we did this experiment for understanding the process of protein
denaturation and coagulation to determine the effects and compare the differences by observing.

Purpose
To observe the processes of protein denaturation and coagulation by doing three
experiment - those are with milk with vinegar, soy milk with Magnesium Sulfate, and raw
chicken with heat.

Material
1.
2.
3.
4.
5.

4 Beakers
120 mL milk
Hot plate
11 mL vinegar
120 mL Soy milk

6. 1.61g. Magnesium Sulfate

7. 2 pieces of cloth
8. 2 pieces of raw chicken
9. Microscope
10. Massing Paper
11. Petri dishes

Procedure
Experiment 1: Precipitate Casein From Milk with an Acid (Vinegar)
1. Weigh the empty beaker and record the weight.
2. Weight and record the weight of 120 mL of milk in the beaker. Find the weight of
the milk only.
3. Place the beaker with milk on a hot plate. Heat it. Remove the beaker.
Add 11 mL of vinegar to the warm milk and stir for 2 minutes. Allow to sit for 5 minutes.
The casein will precipitate into heavy white curds.
4. Obtain a piece of cloth and cover the top and sides of a beaker. Pour the curdled
milk into the beaker, collecting the curds (casein) in the cloth and allow the vinegar and
whey to drain off into the bottom of the beaker.
5. Gather up the cloth with the casein and rinse in cool water by dipping into another
beaker containing water.
6. Squeeze the casein until almost dry, then spread out the cheesecloth to let the
casein dry for 5 minutes.
7. Weigh the precipitate of the curds without the cloth. Record your results.
Experiment 2: Coagulation of Protein from Soy Milk Using a Salt
(Magnesium Sulfate)
1. Weigh and record empty beaker. Weigh and record the weight of 100 ml of soy
milk in the beaker. Record the weight of soy milk only in your data table.
2. Place beaker with soy milk on hot plate. Bring the soy milk to a boiler and turn
off the heat.
3. Add 1.61 grams of magnesium sulfate to paper, then fold the paper to pour into
hot soy milk in the hot plate and stir it.
4. Wait until the curds are floating in an almost clear liquid.
5. Obtain piece of cloth and cover the top of beaker. Pour the soy curds and liquid
into beaker , collecting the curds into a cloth and allow liquid to drain into bottom of
beaker.
6. Gather up the cloth and squeeze out as much water as possible. Spread out the
cloth and allow the curds to dry for 5 minute
7. Weigh the curds without the cloth. Record your results.
Experiment 3: Determining the Effect of Heating Proteins
by Comparing Raw versus Cooked Meat
1. Obtain 2 pieces of raw meat.
2. Put one into a beaker of boiling water on a hot plate.

3.
4.
5.
6.

After the meat is cooked, take it out and let it cool down.
Put it on the petri dish both raw and cooked
Put it under the microscope and observe the differences between these two.
Record your results.

Data Table

Data Table 1: Milk and Soy milk curds

Weight of Empty Beaker: 102.12 g., 102.62 g.
Food Type

Weight of milk/soy
milk

Weight of curd

Percent of the curd

comparing to the
solution

Milk + Acid
(Vinegar)

120.3 g.

18.94 g.

15.74%

Soymilk + Salt

88.86 g.

21.81 g.

24.54%

Data Table 2: Comparison of Meat texture with Microscope

Raw Chicken
Stretchable
Flexible
Soft
Slimey

Cooked Chicken
Shrinkable
Color changed
harder
can be pull out in small pieces

Analyzing Results
1. Describe the difference in textures between the raw chicken muscle and the
cooked chicken muscle.
The textures between the raw chicken muscle and the cooked chicken muscle seem to
have lots of differences. The raw chicken muscle have a soft looking surface, light flesh tone
colored, little slimy and also flexible. On the other hand, the cooked chicken muscle shrink a bit
and the color changed after it was boiled. When we touched it, it didnt look stretchable
anymore, it can be pull out into a small pieces like fiber.

Raw chicken

Cooked chicken

2. Compare the weights of the curds from the milk with that from the soymilk.
Explain why you think theres a difference.
The we got 18.94g of curds from the milk with vinegar (acid) and 21.81g of curds from
the soymilk with Magnesium Sulfate (salt). Milk that are consist of casein, whey protein and
other proteins has more proteins than soymilk, which lead us to have more curds from milk than
the soy milk (Columbia Health, 2012). Unfortunately, we got more amount of curd from soymilk
than the milk. We thought that there are differences between these two kinds of protein because
we did not really use the same amount of the solution as mentioned in procedures. The beaker
have wide range about 50 ml which make it very hard for us to get both exact amounts following
given procedures and lead to the different weights of both curds. Additionally, The chemicals that
we added for the solution to denature and coagulate were different. Thus, the weights of milk and
soymilk were completely different.

Milk curds
Soy milk curds
3. What percent casein from milk was your group able to collect? How about soy
protein from soy milk? Which group in your class had the highest percentage?
Food type
Milk + Acid (Vinegar)

The highest percentage

Nons group : 20.39%

Our Percentage
15.74%

Soymilk + Salt

Pews group : 30.1%

24.54%

We were able to collect 15.74% of the curd from 120.3g of milk and 24.54% from 88.86g
of soymilk. None of our results are one of the highest results in our class.
Nons group have the percentage of 20.39%. Which they extracted the highest amount of
curds from the Milk.Pews group have the highest amount of curds from the Soy milk of 30.1%.
The difference results might be caused by the level of temperature, the amount of
chemical solution, and some processes, such as squeezing unequally.
4. Describe the process of denaturation and coagulation of proteins.
Denaturation and Coagulation are the processes that protein loses its structure by
independent factors such as heat and chemical. (Derrick Arrington, n.d.) The protein that was
denatured could change its characteristics and appearance because of the breaking of hydrogen
bonds. Also, other factors such as pH, acid, bases, and organic or inorganic molecules. Denatured
proteins can show the wild changes due to the loss of protein condensed, and affecting the
dissolution. Most function lose their abilities. For the example, the enzyme activity was also
being denatured. Enzyme lose their function and shape, especially active sites which affect the
whole function mainly the binding of the substrates for the processing. Protein denaturation are
rarely reversible, the process of reverse calls renaturation. One example that can be reverse is
eggs, egg white can be returned by using vitamin C or sodium borohydride. When the protein
denaturation happened, the coagulation continue next.

Coagulation is a process that change the state of the substances from liquid into solid
mass. Then, forming a clot/curds. According to the milk and soy milk experiment, either milk or
soy milk. In the process of coagulation, both solutions start to clot and forming white and solid
substances from protein in milk and soy milk. (The Editors of Encyclopdia Britannica, n.d.)

5. Research why adding the vinegar to milk denatures the casein proteins. Describe
the pH change that takes place.
The pH of milk is slightly acid (less than 7). The role of vinegar is to reduce the pH of
milk into the optimal level. There would be casein proteins separated from the solution and make
them change their state and shape. The molecules of the milk changes into a new shape after the

cool-down process which make them stick together, making a white thick substances or we can
call it a curds.

6. Coagulation of blood takes place in the human body. Link you knowledge of
coenzymes from biology class to this process.Research which coenzymes are needed in
order for blood to coagulate, and briefly explain how it work.
Vitamin K is the cofactor which is very important in blood clotting. It makes blood clots
work normally. There is no limit of intake of vitamin K we should follow yet but somehow too
many blood clottings are not good for our health. Deep Vein Thrombosis or (DVT) is one of the
issue we may have from blood clots. To prevent blood clots theres one substance in our body
which can help prevent the blood clot formation, a coenzyme called Coenzyme Q10. Its an
antioxidant, fighting with the free radicals in our body. It work likely to the vitamin K, increase
the risk of bleeding if took with some certain drugs.

Conclusion
In our three experiment, we have learned about information of proteins and their
processes, those are denaturation and coagulation. We did this experiment by using soymilk, milk
and raw chicken to demonstrate these processes, and using chemical solutions and heat as the
factors to interact the processes. The protein denaturation occurred by adding some chemical
solutions and changing temperature, which causes protein unfolded. Then, the unfolded proteins
coagulate and pack to form the soft semi-solid clot. These terms of proteins cannot be
irreversible that means it cannot be similar to original in terms of state, characteristic and
appearance. These three experiment showed us the different results of denaturation and
coagulation. For the milk experiment, we got 15.74% of curds from 120.3 g. of milk by adding
vinegar as the acid in order to stimulate the interaction. And for the soymilk experiment, we got
24.54% of curds from 88.86g. by adding Magnesium Sulfate as the salt. The different between
results of milk and soymilk experiment might occur by the type of proteins in curds. The protein
in milk curds is casein, but protein of soy milk curds is soy protein. Therefore, the curds from
soymilk were soy protein that might have more concentration and amount of protein than casein.

It caused the different between the percentage of curds. Moreover, other reason that we got the
higher percentage of curds from the soymilk than milk might be because we did not squeeze all
of the water remaining in the curds similar to what we did to the curds from soymilk. And
another experiment was heating the raw chicken. We observed that its texture and appearance
were changed because of the temperature. While we were boiling the chicken, the cooked
chicken was shrinked, changed color and could tear it to be small pieces like the fiber of
muscles. In addition, for the cooked chicken, we saw a small piece of hair when we observed it
through the microscope. We predicted that we did not do this experiment neatly enough. Thus,
some of the hair might stuck with both pieces of chicken meat. However, the tiny hair did not
have any impacts on the result of both raw and cooked chicken.
The results of our group were not the highest or the lowest compared to the others. Each
group may pour different amount of solutions into the beaker was the reason why the amount of
curds that were extracted was different, and the amount were not really accurate, it depended on
the amount of liquid that was squeezed out. In addition, while we were pouring Magnesium
Sulfate, we put the beaker of soymilk on the table instead of on the hot plate. As a result, the
curds would not be produced because there is not enough heat to run and speed up the process.
Although, there were the diverse results, we could conclude the fact that when the protein had
denatured and coagulated, it cannot return like the original one.
If we got a chance to redo these experiments, we would make sure that we pour the solutions
following given procedure accurately. Therefore, it would be clearer to us that which one can be
able to have more amount of curds than others. And, we would make sure that we squeeze all the
water from the curds because we felt like we did not squeeze all of the water out. Hence, the
results would not be totally correct. We will try to make it better next time. If not, we will use
these mistakes as the lessons and make sure we will do everything as instructed. We hope to have
the successful experiment one day.

References
Coenzyme Q10. (n.d.). Retrieved April 3, 2015, from http://www.mayoclinic.org/drugssupplements/coenzyme-q10/interactions/hrb-20059019
Nutritional differences between soy- and cow's milk. (n.d.). Retrieved April 3, 2015, from
http://goaskalice.columbia.edu/nutritional-differences-between-soy-and-cows-milk

Denaturation of Protein: Definition, Causes & Quiz | Study.com. (n.d.). Retrieved April 3, 2015,
from http://study.com/academy/lesson/denaturation-of-protein-definition-causesquiz.html
Coagulation | of blood. (n.d.). Retrieved April 3, 2015, from
http://global.britannica.com/EBchecked/topic/69750/coagulation
_______ (n.d.). Retrieved April 3, 2015, from
http://www.teachnlearnchem.com/Matter_and_Energy/PDF/casein glue.pdf
Coenzyme Q10. (n.d.). Retrieved April 3, 2015, from
http://umm.edu/health/medical/altmed/supplement/coenzyme-q10

Work log
Kam

Gam

June

Mind

Preme

24/03/15

Data Table

Analyzing
Results

Material and Conclusion

add pictures

Introduction

31/03/15

Analyzing
Results

Analyzing
Results

Procedure
and
Analyzing
Results

Conclusion

Introduction

02/04/15

Revise

Revise

Revise

Revise

Revise