Preparation, Purification and Identification of

Lysozyme (Part One)

Creative BioMart

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Experiment Summary
Preparation, Purification and Identification of Lysozyme

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Experiment Principle

Lysozyme, the full name of which is 1,4--N-lysozyme, is also known as

muco-peptide N-acetylmuramyl hydrolase, belonging to -lactalbumin
family. The study of lysozyme began in the early last century, when the
British bacteriologist Fleming found that human saliva and tears contains
enzyme that dissolve bacteria cell walls, and because of its bacteriolytic
effect, so it was called lysozyme; thereafter people found the presence of
lysozyme in the micro-organisms, plants, invertebrates and higher animals
among which egg white lysozyme research and application has been quite
deep and extensive. In addition, with the development of genetic
engineering technology, the research on the production of human lysozyme
by microbial fermentation by means of artificial synthesis of genes is also
being carried out in depth.

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Egg white lysozyme, a kind of alkaline globulin, is composed of 129
amino acid residues, and the molecular weight of 14,388. The
molecule contains four pairs of disulfide bonds. Because of the
number of basic amino acid residues is more than the number of
acidic amino acid residues, so its isoelectric point is up to about 11
around 10.5 to 11, and the optimum temperature is 50 . The
optimum pH is about 6 to 7. The amount of lysozyme in egg white
accounted for about 3.4% ~ 3.5% of the total amount of protein.

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Lysozyme is both medicine and health care products, or ideal drug
enzyme in the biochemical drugs. It has been utilized in
pharmaceutical preparations, food and beverage additives,
biochemical reagents and medical diagnostic agents. Lysozyme is
widely used in medicine, food preservation and bio-engineering.
Lysozyme is a glycoside hydrolase, having anti-bacterial anti-
inflammatory, anti-virus, enhance immunity and other
pharmacological effects. It has been clinically used in various ENT
inflammation, flat warts, molluscum contagiosum, common warts
and other skin disease treatment, and also for the prevention and
treatment of viral hepatitis, especially for post-transfusion hepatitis
and acute hepatitis more significant effect. In addition, human
lysozyme can also be used as a diagnostic indicator of various
diseases.

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Main Reagent

Solid ammonium sulfate; 10% ammonium sulfate; 1 mol / L HCl; 1

mol / L NaOH; polyethylene glycol; 0.15 mol / L phosphate buffer at
pH 6.5; 0.1 mol / L HCl; 0.025mol/L KCl-0.1mol/L HAc; 30% Acr/Bis;
10% SDS; 1.5mol/L Tris-HCl (pH 8.8); 0.5mol/L Tris-HCl (pH 6.8);
TEMED; 10% ammonium persulfate; Tris-Gly (pH = 8.3)
electrophoresis buffer; sample processing solution; staining solution;
decolorization solution

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Major Equipment

Precision pH paper; Buchner funnel; magnetic stirrer; centrifuge; dialysis

bag; glass column; chromatography system; vertical electrophoresis
system

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Experimental Materials

A few fresh eggs; 724 acid cation exchange resin; Sephadex G-

50; blue dextran -2000

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Experiment Procedure

To be continued...

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 Thanks for watching!
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