Extraction and Purification of Lysosome from Egg White

Discussion:
Young children are very much receptive to you are sensitive reactions caused by the consump-
tion of egg. The Cytoplasm of the unfertilized egg is known as egg white or albumin. More than
20 types of proteins are present in egg white1. Among the proteins the allergens are,
1. Ovalbumin (Gal d 2)
2. Ovomucoid (Gal d 1)
3. Ovotransferrin (Gal d 3)
4. lysozyme (Gal d 4)
The lysosome is the catalytic enzyme which is known to digest the cell wall of the bacteria and
found abundantly in the albumin or egg white2. The lysosome is the important element present in
the egg white of hen and very much resistant to hydrolysis with Pepsin. The Ion exchange chro-
matography is utilized to extract lysosome from egg white is the objective of the experiment and
verifying the purity of lysosome with the help of molecular and biochemical techniques is the
main objective of conducting this experiment.
The lysosome is first extracted from the egg white and it is purified with the help of size exclu-
sion chromatography. The column in the stationary phase consists of cross-linked gel Matrix
which is porous in nature. The size exclusion chromatography which is utilized for separation
and purifier in the lysosome is dependent on the size of the molecules3. The large molecules
elute at the initial mobile phase but the small molecules are trapped with the beads and they elute
at the end. The different types of protein present in albumin elute at different fractions because
of the size of the molecules to the purification process of the lysosome. Hypothetically it is as-
sumed that lysosome is small in comparison with the fractional range of the column which leads
1 Chen, C., Li, X., Yue, L., Jing, X., Yang, Y., Xu, Y., ... & Zhang, X. (2019). Purification and characterization of
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to trapping of lysosome molecules in the resin beads4. This will result in less amount of the lyso-
some.
One of the most powerful media in the experiment is the enzymatic reaction which is utilized for
the detection of the presence of lysosome in egg white. The lysosome is composed of 128
residue of amino acids which are folded and form a globular structure5. When a substrate clamps
to the left of the structure tend to hydrolyze the peptidoglycan polysaccharide available with bac-
terial cell walls and results in the osmotic lysis of the cell walls. It is a well-known fact that the
Gram-Positive Bacteria are more susceptible to the lysosome because of their exposure to the ex-
tracellular environment6. The Gram-Negative bacteria have a thin layer of peptidoglycan There-
fore they are less susceptible to the lysosome. They have an outer membrane of lipopolysaccha-
ride which Shields them from being hydrolyzed.
The Gram-Positive Bacteria is taken as a sample to be used as a substrate to detect the activities
of lysosome as an enzyme. The presence of lysosome in comparison with the other major pro-
teins in albumin is detected with the help of lysosome assay. The lysosome is very important for
the research because it has the valuable capability of laying the Gram-Positive Bacteria. Like
other biomolecules, lysosome cannot be found in nature. They can only be extracted from albu-
min and purified for further use. The method of extracting lysosome from albumin or egg white
is a simple process and does not require complicated apparatus and method of extraction. The
process of extraction is also not very time consuming, therefore, it is one of the most popular
ways to get lysosome for various use in biochemistry.
It is a known fact that lysosome is very much resistance to the hydrolysis with pepsin but with
the help of various experiments that can be stated that lysosome can be hydrolyzed with pepsin
when the pH is 1.2. Low pH is chosen to hydrolyze lysosome with pepsin in the simulated gas-
tric fluid. Optimal conditions are very effective and important to find the appropriate result. The
substrate concentration assays, as well as the pH essay, are used in this experiment to understand
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the enzyme activity of lysosome as those activities vary with the change in the concentration and
buffer pH7. It is very important to have the appropriate concentration as the higher concentration
of the substrate.
There is a similarity on the binding properties of lysosome and the lysosome contaminated pro-
teins; therefore, it is very essential to determine the activity of lysosome using the micrococcus
suspension8. The amount of micrococcus which is appropriate for substrate concentration of
lysosome activity of 250 units is 0.4 mg per ml. It is very important to determine the optimal pH
where the bonding between the substrate and the enzyme is most efficient it9. If an environment
is too basic or too acidic it can cause obstacles for the bonding between the enzyme and the sub-
strate which results in the low activity of the lysosome. In the lab, it is proved that pH 7 is the
ideal pH for the activities of lysosome can be carried out very effectively with the enzyme activ-
ity of 300 units.
Conclusions:
In the experiment, lysosome is successfully extracted from the album in out the egg white and it
is also purified using the column chromatography at the pH of 7. The concentration of the lyso-
some which is very essential to determine in this experiment is successful to determine the purity
of lysosome with the help of lysosome assay. The purity of lysosome is confirmed with the help
of SDS page waste Assay. From the experiment, it can be successfully concluded that column
chromatography is very useful to extract lysosome from egg white because like other enzymes
lysosome is not available in nature and it should be extracted from the source10. Purification of
the lysosome is also very necessary procedure as the contaminated lysosome cannot be utilized
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