PROTEIN DENATURATION

DENATURATION – is the process of unfolding and rearrangement of the secondary and

tertiary structure of protein causing it to lose its biological function.

– Proteins are coagulated by heat, strong acids, alcohols, some metallic ions like mercury
ion (Hg2+), lead ion (Pb2+) and silver ion (Ag+) and alkaloidal precipitating agents like tannic
acid and picric acid.

A. COAGULATION BY HEAT, ACIDS AND ALCOHOL

1. HEAT
Put a pinch of powdered egg albumin in each of 2 test tubes labeled no. 1
and no. 2. To test tube no. 1, add 2mL distilled water then place both test
tubes in a boiling water bath for 10 minutes, with constant shaking. Remove
the test tubes, cool to room temperature and then add 2mL of distilled water
to test tube no. 2. Filter the solutions and test bothe filtrates with Biuret
reagent. Compare the results.

2. (A) INORGANIC ACIDS

Prepare 2 test tubes containing 1mL each of egg albumin solution. To test
tube no. 1, add conc. HCl, dropwise until a precipitate is formed, counting the
number of drops of the acid for precipitation to occur. Then add excess of HCl
and take note whether it will increase or dissolve the precipitate formed.
Repeat the same procedure in test tube no. 2, but this time use conc. H 2SO4.

(B) HELLER'S TEST

Place 1mL of egg albumin in a test tube. Hold the test tube in an inclined
position, and then add 4mL concentrated HNO3 slowly along the side of the
test tube. DO NOT SHAKE.

3. ALCOHOLS
Place 1mL of egg albumin solution and 1% gelatin solution in separately
labeled test tubes, add 5mL of 95% ethanol in each test tube, mix thoroughly.
Observe the formation of precipitate. Test the solubility of this precipitate in
water.

B. PRECIPITATION OF PROTEINS

1. BY HEAVY METALS
Place 2mL of egg albumin in each two test tubes. Add the following reagents
to the test tube drop by drop with shaking.

Test Tube 1: 1% Lead Acetate Test Tube 2: 1% HgCl2

 Count the number of drops needed to produce a precipitate.

2. ALKALOIDAL REAGENTS
Place 3mL of egg albumin solution into two test tubes. In test tube 1, add 2mL
of tannic acid solution, and in test tube 2, add 2mL picric acid solution.
Describe the protein solution in each test tube after addition of alkaloidal
reagents.