Denaturing Proteins

Objective: To experiment with different methods of denaturing albumin, the protein found in
egg white.

Materials:
Hot Plate 400 mL beaker 6 test tubes
Egg white NaCl NaOH
HCl 1% AgNO3 Ethanol
Stirring Rods Graduated cylinder

Background: Proteins are large molecules made up of small amino acids. Proteins are held in a
natural shape due to interactions of the R groups on the amino acids. We can denature a
protein by disrupting these interactions. When a protein is denatured the overall shape of the
protein changes and new properties can be observed. The shape of a protein is associated with
food processing properties, such as solubility, gel formation and enzyme activity.

We will explore how the following denature albumin:

 Heat Albumin
 Acids and Bases
 Organic compounds
 Heavy metals

Procedure:
1. Label 6 test tubes 1 – 6
2. Fill the test tubes with 5 mL of albumin – does not need to be perfectly exact.
3. Place test tube 1 in a boiling water and allow to “cook” till egg turns white.
4. Add a large pinch NaCl to test tube 2 and stir.
5. Add 5 drops of NaOH to test tube 3 and stir.
6. Add 5 drops of HCl to test tube 4 and stir.
7. Add 1 mL of ethanol to test tube 5 and stir.
8. Add 5 drops 1% AgNO3 to test tube 6 and stir.
9. Record observations in a table in your lab notebook.
10. Dispose of the silver nitrate test tube in the waste container.
11. All remaining test tube contents can be poured down the drain.
12. Please wash and rinse your test tubes. DO NOT DRY!

* Some of these reactions will take time, so be patient. Give steps 4 – 8 some time to react.
Post Lab Questions: (Answer your questions in complete sentences in your lab notebook)
1. In your own words, explain the effect of heat on albumin.

2. Did you observe any differences in the albumin at a pH of 2 (HCl) and a pH of 13 (NaOH)?
What may have caused these changes?

3. What kinds of disruptions to the conformation occur in each of the following? In other
words, are hydrogen bonds, disulfide bridges, etc. disrupted?
a. Which of the interactions below would be disrupted by heat?
b. Which of the interactions below would be disrupted by an alcohol?
c. Which of the interactions below would be disrupted by metal ions?

NH2 OOC Ar
N H S S Ar
O C

C O H N

Hydrophobic
hydrogen bonds salt bridge disulfide bridge Interaction

4. When milk sours, lactic acid is produced and a white precipitate forms. What is the
precipitate? Explain what is happening.

5. Ceviche is a seafood dish popular in the coastal regions of Mexico, Ecuador, Colombia, Chile
and Perú and other parts of Latin America. It contains different types of fish and lemon or
lime juice. The fish is never heated but technically it is cooked. Explain how the fish is cooked.