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A protein is a naturally occurring, unbranched polymer in which the monomers
units are amino acids, which at least has 50 amino acids residues. The arrangement
of these amino acids contributes in the unique physico-chemical characteristics of a
protein which determine what protein isolation and purification method should be
done. To denature a protein is to break its native conformation by subjecting the
protein to extreme heat and pH, introducing chaotropic agents, or infusing it with
chemicals that can break disulfide bonds. To isolate casein from skimmed milk, an
acid is used to adjust the pH to 4.6 which is the isoelectric pH of the casein. The
isolated casein has undergone acid hydrolysis; this process is done to obtain
information about the isolated casein’s composition, then, protein is treated with an
acid, alkali, or proteolytic enzymes. For Qualitative color reactions, after auto-
claving, the intact proteins were used in different tests such as Biuret test,
Ninhydrin test , Xanthoproteic test , Millon’s test, Hopkins-Cole test, Sakaguchi
test, Nitroprusside test, Fohl’s, test for amides, and Pauly’s test. Paper
chromatography was performed for the separation and identification of amino acid
standards based on the polarities of tryptophan, arginine, proline, cysteine, serine,
aspartate, histidine, glycine, and alanine.