PROTEIN EXTRACTION

Good Samaritan Colleges

A Laboratory Report by:

Rebolledo, Nadine P.

Science, Technology, Engineering, and Mathematics (STEM)

Group 2 of 12 STEM 7

27 January 2023

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INTRODUCTION

In 1838, Jöns Jakob Berzelius discovered proteins, which is why we further learn and

understand the life that embodies us. Proteins are macromolecules, polymers of amino acids, where

thousands of amino acids are attached to one another in long chains to form a protein (Blanco &

Blanco, 2017). Moreover, proteins are one of the most actively studied molecules in biochemistry.

They work inside cells and form the basis of living cells, muscles, and tissues required for the

structure, function, and regulations of the body's organs (Wilke, 2020). Thus, our body would not

function properly to build, repair, and support muscle tissue without proteins.

Protein helps produce antibodies that prevent diseases and infections and create new

cells (Cooper, 2022). Moreover, more types of protein are on the market, including extracted ones,

such as casein proteins from milk. According to Walle (2018), casein makes up 80% of milk

protein and the other 20% of whey protein. It is a heterogeneous mixture of phosphorous-

containing proteins in milk, with high-quality proteins that contain all essential amino acids

required to nourish one's body. Casein can be present in milk in two different forms: calcium salt

and calcium caseinate. The micelle cluster known as casein is made up of alpha, beta, and kappa

caseins. The milk's opaque white appearance is due to these micelles (Virtual Amrita Laboratories

Universalizing Education, 2011). Casein can be separated from milk by bringing it to its isoelectric

point through acidification.

The number of the protein's negative charges equals the number of positive charges at

the isoelectric point. Proteins tend to aggregate through electrostatic interactions, making them

least soluble in water at their isoelectric points (Pollock, 2016). The positive end of one protein

molecule lures the negative end of another protein molecule, and the aggregates precipitate out of

the solution.

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OBJECTIVES

• Isolate casein from milk.

• Provide a thorough record and understanding of the extraction experiment.

• Analyze and present conclusions based on the experimental work.

MATERIALS AND METHODS

MATERIALS

• Powdered Milk

• Distilled Water

• Acetone

• Ethyl Alcohol

• White Vinegar

• Cheesecloth

• Measuring Cup (with graduation)

• Thermometer

• Bond Paper

• Stove

• Pot

• Stirrer

• Small Jar

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PROCEDURE
(Schematic Diagram)

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RESULTS, DISCUSSION, AND DOCUMENTATION

Results

Procedure / Step Observations Explanation /

Purpose of this step

Addition of water to Adding water to powdered To dilute the powdered milk

powdered milk milk significantly affects its and become a uniform color
consistency; over time, it change.
dissolves as we keep stirring.

Addition of vinegar The casein slowly separates To reduce the PH of the milk
from the solution. and, at the same time, for the
protein casein to reach its
isoelectric pH.

Addition of ethyl alcohol Added ethyl alcohol filtrates It lowers the dielectric
extra fat and dissolves it into constant of the electrostatic
(ethanol) the solvent mixture. interactions that stabilize the
casein.

Addition of acetone Acetone breaks down and The addition of acetone

dissolves other residue allows the precipitation of
substances, such as fat and peptides or proteins.
phospholipids, directly from
freeze-dried casein.

Discussion

1. What is the name of the protein extracted?

• Casein Protein

2. Why vinegar makes the milk coagulate?

• Since vinegar is an acid, and acid lowers the pH level of milk, it coagulates. The changes

in milk's pH make casein molecules evolve and reorganize into a long chain, which curdles

the milk (Potenza, 2017).

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3. Write the chemical name of vinegar, draw its structure, and determine if it is a strong or

weak acid.

• Chemical Name of Vinegar: Acetic Acid (CH3COOH)

• Weak acid refers to an acid that partially ionizes in an aqueous solution (LibreTexts

Libraries, 2022). Thus, vinegar or acetic acid is weak as it only partially dissociates when

placed in an aqueous solution, such as water.

4. What is the purpose of ethyl alcohol?

• Its purpose is to lower the dielectric constant of the electrostatic interactions that stabilize

the casein. It filtrates and dissolves extra fat into the solvent mixture when added to the

experiment.

5. What is the purpose of acetone?

• Acetone breaks down and dissolves other residue substances directly from freeze-dried

casein. It allows the precipitation of peptides or proteins.

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Documentation

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Documentation

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CONCLUSION

Casein is the primary protein present in milk. It is used in processed foods and other

industrial products. Through isoelectric precipitation, casein was extracted from milk as it exists

as the calcium salt and calcium caseinate. The isolated casein was subjected to acid, which caused

visible changes comparing the isolate formation before and after extracting it; by lowering the pH

of the milk to its isoelectric point, it is possible to extract casein as most proteins exhibit the lowest

solubility at this point. In the isolation of the said experiment from milk, the precipitation of protein

that is soluble in water and a group of proteins soluble in salt solutions can be observed.

REFERENCES

Blanco, A., & Blanco, G. (2017). Medical Biochemistry. In A. Blanco, & G. Blanco, Medical
Biochemistry (pp. 21-71). Elsevier.

Cooper, J. (2022, September 21). Benefits of Protein. (M. Juber, Editor) Retrieved February 1,
2023 from Nourish by WebMD: https://www.webmd.com/diet/benefits-protein

Lawandi, J. (2015, August 27). The Science of Why Acid Curdles Milk. From ktchn:
https://www.thekitchn.com/the-science-behind-why-acid-curdles-milk-222962

LibreTexts Libraries. (2022, May 1). Strong and Weak Acids and Acid Ionization Constant. From
LibreTexts Chemistry:
https://chem.libretexts.org/Bookshelves/Introductory_Chemistry/Introductory_Chemistry
_(CK-
12)/21%3A_Acids_and_Bases/21.12%3A_Strong_and_Weak_Acids_and_Acid_Ionizati
on_Constant_(K_texta)#:~:text=A%20strong%20acid%20is%20an,slightly%20in%20an
%20aqueous%20so

Pollock, E. (2016, January 5). Isolation and Identification of Casein. From Stockton Wordpress
Blogs: https://blogs.stockton.edu/chemlifesci2/files/2016/01/casein.pdf

Potenza, W. (2017, July 18). Why does milk curdle when it is mixed with vinegar? From Linked
In: https://www.linkedin.com/pulse/why-does-milk-curdle-when-mixed-vinegar-walter-
potenza#:~:text=Milk%20curdles%20when%20mixed%20with,of%20protein%2C%20su
gar%20and%20fat.

Virtual Amrita Laboratories Universalizing Education. (2011). Isoelectric Precipitation of

Proteins: Casein from Milk. Retrieved January 1, 2023 from Amrita Vishwa
Vidyapeetham: https://vlab.amrita.edu/index.php?sub=3&brch=63&sim=158&cnt=6

Wilke, C. (2020, June 1). Scientists Say: Protein. From Science News Explores:
https://www.snexplores.org/article/scientists-say-
protein#:~:text=protein%3A%20A%20compound%20made%20from,the%20work%20in
side%20of%20cells.

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