Experiment 7: Protein
Introduction
Conformation of protein is resulted from the inter-protein interactions that will hold the
molecule in a given conformation while protein aggregation is related with intra-protein
interactions. The most important of these here are:
Hydrophobic interactions. Hydrophobic amino acids will try to avoid water by
either coiling into the core of the polymer or adsorbing into a non-polar solvent. The
hydrophobic interactions of protein can be adjusted by denaturing the protein.
Electrostatic interactions. Proteins have ionizable amino acids that may carry a
positive or negative charge depending on the pH.
Globular proteins can be thermally denatured by heating, then the globule unfolds and
exposed some of the hydrophobic core amino acids to the aqueous solvent. If the structure
does not regenerate, there will be a pressure to aggregate to reduce the hydrophobic
interaction. Based on these simple rules of protein behavior we can try to understand the
functional properties of whey proteins in film.
To form a solid gel, a protein must aggregate and form a continuous structure throughout
the container. This can be seen as a partial precipitation of the protein because there are
strong protein-protein interactions.
Many proteins can stabilize emulsions because their hydrophobic amino acids partition into
the surface of the freshly formed oil droplet and cause the protein to stick at the oil-water
interface (i.e., adsorb). The adsorbed layer gives some stability by shielding the oil from
the aqueous phase but the proteins now to a great extent control the functional properties
of the system. If the protein tends to aggregate (e.g., following thermal denaturation or in
the absence of strong electrostatic repulsion), the droplets they are attached to will
aggregate also. Aggregation of the droplets in an emulsion can lead to gelation, but can also
lead to creaming because the flocculated droplets are much larger.
Milk protein consists of 80% casein and 20% whey protein. There are four major types of
casein molecules: alpha-s1, alpha-s2, beta, and kappa. Milk, in its natural state, is
negatively charged. The negative charge permits the dispersion of casein in the milk.
Precipitation of casein can be done using acid or rennin.
When an acid is added to milk, the H+ concentration neutralizes the negatively
charged casein micelles. At pH 4.7, the isoelectric point (the point at which all
charges are neutral) of casein, an isoelectric precipitate known as acid casein is