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Erika C. Rabara, Harvey Mher M. Rarang, Maika L. Regala, Chelejan Mhare U. Regino, Erik
Kristian Victor B. Sabio, Mica Gienela A. Sanchez, Arianne Nicole Denise T. Yoro
Group 8
2D Medical Technology
Biochemistry Laboratory
ABSTRACT
Myoglobin is the primary oxygen-carrying pigment of muscle tissues. It contains hemes, pigments responsible for the
color of red meat. The color that meat takes is partly determined by the degree of oxidation of the myoglobin. This
portion of experiment refers to the denaturation of intact myoglobin through hydrolysis. The pre-isolated myoglobin
from meat sample was subjected to basic hydrolysis to alter its native conformation and thus making a suitable
hydrolysate for qualitative and quantitative characterization. NaOH was used as a medium in basic hydrolysis and HCl
was used to neutralize the mixture while in acid hydrolysis, HCl was the medium used and NaOH was the neutralizing
agent. It has been concluded that in hydrolysing proteins, basic and acid hydrolysis is effective and efficient.
INTRODUCTION
Myoglobin
is
a single-chain globular
protein of 153 or 154 amino acids, containing
a heme
which
is
an
ironcontaining porphyrin prosthetic
group in
the
center
around
which
the
remaining apoprotein folds. Biologically,
active
proteins, like myoglobin, are made up of
polymers consisting of amino acids linked by
covalent peptide bonds. These bonds are broken
when the protein undergoes hydrolysis. In
hydrolysis, the protein is subjected to extreme
conditions usually at high temperatures by
prolonged boiling in a strong acid or strong
base or
using
an enzyme such
as
the
pancreatic protease enzyme to stimulate the
naturally occurring hydrolytic process. This will
cause denaturation of the protein meaning that
the proteins conformation is altered by the
breaking of peptide bonds. This results to a
solution containing amino acid fragments which is
then called the hydrolysate. Denaturation alters
protein function, demonstrating a relationship
between structure and function. Hydrolysis of
protein and analysis of products are done to
obtain information about their compositions. The
three most common types of hydrolysis are aid,
basic and enzymatic hydrolysis. Acid hydrolysis
implies
a
chemical
mechanism
of hydrolysis catalyzed
by
a Brnsted or Arrhenius acid. By contrast, it does
not usually imply hydrolysis by direct electrophilic
attackas may originate from a Lewis acid.
Nevertheless, the type of hydrolysis carried out
with a basic medium is termed basic hydrolysis.
Lastly,
an
enzymatic
hydrolysis
is
by
the addition of specific enzymes called proteolytic
enzymes or simply proteases which refer to a
group of enzymes each to hydrolyse specific
peptide bonds of proteins. [1]
EXPERIMENTAL
A. Compounds tested (or samples used)
Buffered muscle extract and 70% bufferdiluted (NH4)2SO4 solution form crystals was used
for the isolation of myoglobin.
The reagent used for alkaline hydrolysis
was 4M NaOH; and 1M HCl was used for
neutralization purposes. Blue litmus paper was
also used to test the pH of the solution.
B. Procedure
1. Isolation of Proteins
REFERENCES
[1] Biochemistry of muscle, muscle contraction.
Biochemistry of connective tissue. (2013).
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