ISOLATION AND PURIFICATION OF LYSOZYME FROM THE HEN EGG WHITE

The aim of the research was the development of the method of lysozyme isolation from hen egg proteins. Lysozyme was isolated by differential heat denaturation of proteins with changing of the medium pH value, followed by neutralization, dialysis and additional purification by gel chromatography on S...

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Main Authors: S. S., I. I., A. M., M. G., V. A.
Format: Article
Language:English
Published: National Academy of Sciences of Ukraine, Palladin Institute of Biochemistry 2015-12-01
Series:Biotechnologia Acta
Subjects:
Online Access:http://biotechnology.kiev.ua/images/stories/6_2015/dekina_6_2015.pdf
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author S. S.
I. I.
A. M.
M. G.
V. A.
author_facet S. S.
I. I.
A. M.
M. G.
V. A.
author_sort S. S.
collection DOAJ
description The aim of the research was the development of the method of lysozyme isolation from hen egg proteins. Lysozyme was isolated by differential heat denaturation of proteins with changing of the medium pH value, followed by neutralization, dialysis and additional purification by gel chromatography on Sephadex G-50. Activity was determined by bacteriolytic method (with Micrococcus lysodeikticus 4698 as a substrate). The enzyme purity and molecular mass were determined using SDS-electrophoresis and massspectrometry. The method of lysozyme isolation from hen egg proteins with the enzyme yield of 3.2 ± 0.2% and bacteriolytic activity of 22 025 ± 1 500 U/mg is modified. According to electrophoresis data, the isolated enzyme is characterized by high degree of purity (~95–98%) and is comparable with lysozyme of AppliChem company by main physical and chemical characteristics. The obtaining product is stored in a crystalline form at low temperature (–24 оC) for 9 months. The proposed method allows obtaining active and stable lysozyme with high purity from hen egg protein in laboratory conditions for the usage in biotechnology.
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spelling doaj.art-d0ed5907a3ad4a3c80d3ae6842de43fc2023-09-02T12:58:56ZengNational Academy of Sciences of Ukraine, Palladin Institute of BiochemistryBiotechnologia Acta2410-77512410-776X2015-12-0186414710.15407/biotech8.06.041ISOLATION AND PURIFICATION OF LYSOZYME FROM THE HEN EGG WHITES. S. 0 I. I. 1A. M. 2 M. G. 3V. A. 4DekinaRomanovska OvsepyanBodyul ToptikovThe aim of the research was the development of the method of lysozyme isolation from hen egg proteins. Lysozyme was isolated by differential heat denaturation of proteins with changing of the medium pH value, followed by neutralization, dialysis and additional purification by gel chromatography on Sephadex G-50. Activity was determined by bacteriolytic method (with Micrococcus lysodeikticus 4698 as a substrate). The enzyme purity and molecular mass were determined using SDS-electrophoresis and massspectrometry. The method of lysozyme isolation from hen egg proteins with the enzyme yield of 3.2 ± 0.2% and bacteriolytic activity of 22 025 ± 1 500 U/mg is modified. According to electrophoresis data, the isolated enzyme is characterized by high degree of purity (~95–98%) and is comparable with lysozyme of AppliChem company by main physical and chemical characteristics. The obtaining product is stored in a crystalline form at low temperature (–24 оC) for 9 months. The proposed method allows obtaining active and stable lysozyme with high purity from hen egg protein in laboratory conditions for the usage in biotechnology.http://biotechnology.kiev.ua/images/stories/6_2015/dekina_6_2015.pdfhen egg proteinlysozyme
spellingShingle S. S.
I. I.
A. M.
M. G.
V. A.
ISOLATION AND PURIFICATION OF LYSOZYME FROM THE HEN EGG WHITE
Biotechnologia Acta
hen egg protein
lysozyme
title ISOLATION AND PURIFICATION OF LYSOZYME FROM THE HEN EGG WHITE
title_full ISOLATION AND PURIFICATION OF LYSOZYME FROM THE HEN EGG WHITE
title_fullStr ISOLATION AND PURIFICATION OF LYSOZYME FROM THE HEN EGG WHITE
title_full_unstemmed ISOLATION AND PURIFICATION OF LYSOZYME FROM THE HEN EGG WHITE
title_short ISOLATION AND PURIFICATION OF LYSOZYME FROM THE HEN EGG WHITE
title_sort isolation and purification of lysozyme from the hen egg white
topic hen egg protein
lysozyme
url http://biotechnology.kiev.ua/images/stories/6_2015/dekina_6_2015.pdf
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