BIOCHEMISTRY

PROTEIN
Presented by: Group 5
Protein
• Proteins are nitrogen containing organic
compounds composed of amino acid connected to
one another by peptide bonds. Each amino acid
has a basic amino (NH2) group and an acidic
carboxyl (COOH) group, with a characteristic
radical group. The chemical properties of amino
acids are due to these group, and the chemical
properties of the group of the proteins formed
from these amino acids, in turn are dependent on
their chemical properties.
Coagulation Tests
• By Alcohol
- The alcohol test is performed on fresh milk to determine
whether or not it will coagulate during thermal processing.
This test is particularly significant in the production of UHT
milk, evaporated milk, and milk powders. This test has
higher sensitivity than the Clot-on-Boiling (COB) test.
• By Heat
- The heat coagulation test is used to detect albumin and
globulin protein in urine samples. Because albumin and
globulin are found in urine under a variety of clinical
circumstances, their presence may be firmly verified by
this test, which assists in illness diagnosis.
Precipitation Test
• By salts of heavy metals
- Heavy metals such as Ag+, Pb2+, Hg2+, and others
precipitate after forming a compound with alkaline
proteins. Keep track of the amount of precipitation in each
experiment. Organic acids have high negative charges that
neutralize positively charged proteins, resulting in an
insoluble salt.
Precipitation Test
• By strong mineral acids
- The test is based on the notion of protein precipitation,
which occurs in the presence of mineral acids such as
nitric acid. Protein precipitation by acids is dependent on
pH fluctuations in the solution. Because all proteins have a
set isoelectric point, or pI value, changes in the pH of the
solution have an effect on the protein's structure.
Precipitation Test
• By alkaloidal reagents
- Alkaloidal reagents (e.g. tannate and trichloroacetate) are
high molecular weight anions. The negative charge of
these anions counteracts the positive charge of the amino
group in proteins, giving a precipitate.
Color Reaction Tests
• Biuret Test
- The biuret reagent (copper sulfate in a strong base) combines
with protein peptide bonds to generate the "biuret complex,"
which is blue to violet in color. N.B. For this complex to form, at
least two peptide bonds must be present.
• Xanthoproteic Test
- Concentrated nitric acid combines with aromatic rings that are
benzene derivatives to produce the distinctive nitration reaction.
The amino acids tyrosine and tryptophan have active benzene
rings that are readily nitrated to produce yellow compounds.
Despite the presence of a benzene ring, the aromatic ring of
phenyl alanine does not react with nitric acid because it is not
activated.
Color Reaction Tests
• Millon’s Test
- In Million's test, the phenol group of tyrosine is first
nitrated by nitric acid in the test solution. In this case, the
nitrated tyrosine complexes mercury ions in the solution to
form a brick-red solution or precipitate of nitrated tyrosine.
In all cases, the appearance of red color is a positive test.
Glyoxylic acid test (Hopkin’s – Cole Test)
- The Hopkins-Colé reagent (magnesium salt of oxalic acid)
produces favorable findings with proteins containing the
important amino acid "tryptophan," suggesting a high
nutritional value.
Color Reaction Tests
• Ninhydrin Test
- The ninhydrin test is a chemical test used to identify
ammonia, primary or secondary amines, or amino acids. In
this test, ninhydrin reagent is added to the test sample,
resulting in the production of a deep blue hue, also known
as Ruhemann's purple, in the presence of an amino group.
• Sakaguchi test
- The Sakaguchi test is an example of a color reaction test
used to identify amino acids or proteins. The test is an
arginine-specific test in which the guanidinium group of
arginine combines with 1-naphthol or 2-naphthol to
provide a colored product.
Color Reaction Test
• Reduced sulfur test
- When sulfur-containing proteins (such as cysteine and
cystine) are heated with lead acetate in an alkaline media,
a black deposit of lead sulfide (PbS) forms.
Video
Procedure
Coagulation Test by Alcohol
Coagulation Test by Heat
Precipitation Test by salts of Heavy
metals
Precipitation Test by Strong
mineral acids
Precipitation Test by alkaloidal
reagents
Color Reactions Test
Biuret Test
Xanthoproteic Test
Millon’s Test
Glyoxylic acid Test (Hopkin – Cole
Test)
Ninhydrin Test
Sakaguchi Test
Reduced Sulfur Test
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