CN104004731A - Method for preparing lysozyme by egg white - Google Patents
Method for preparing lysozyme by egg white Download PDFInfo
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- CN104004731A CN104004731A CN201410269341.9A CN201410269341A CN104004731A CN 104004731 A CN104004731 A CN 104004731A CN 201410269341 A CN201410269341 A CN 201410269341A CN 104004731 A CN104004731 A CN 104004731A
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- Prior art keywords
- egg white
- preparing
- lysozyme
- diacetylmuramidase
- supernatant liquor
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
- C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
- C12N9/2462—Lysozyme (3.2.1.17)
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
- C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
- C12Y302/01017—Lysozyme (3.2.1.17)
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- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
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- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Engineering & Computer Science (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Enzymes And Modification Thereof (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention discloses a method for preparing lysozyme by egg white. According to the technical scheme, the egg white is used for preparing the lysozyme. The method comprises the operation steps that at a room temperature, the egg white is diluted to 1.5-2 times by a sodium chloride solution, 2 mol/L hydrochloric acid is used for adjusting the PH value to 4-5.5, stirring is conducted for 45 minutes, 6000 r/min rotating speed is used for centrifuging to remove sediments and impurities, liquid supernatant is collected, the liquid supernatant is ultrafiltered by a hollow fiber ultrafilteration membrane with the membrane molecular weight cutoff being 11000 Da, ammonium sulfate is added in the obtained filter liquid to conduct fractional sendimentation on the filter liquid, the liquid supernatant is removed, and the obtained sediments are sprayed with mist and dried. The method for preparing the lysozyme by the egg white has the advantages that the lysozyme is rich in egg white content, simple in extraction process, low in industrial production cost, and suitable for mass production.
Description
Technical field
The present invention relates to N,O-Diacetylmuramidase preparation field, relate in particular to the method for preparing N,O-Diacetylmuramidase with egg white.
Background technology
N,O-Diacetylmuramidase is a kind of alkaline enzyme that can be hydrolyzed mucopolysaccharide in pathogenic bacterium.That this enzyme has is antibacterial, anti-inflammatory, the effect such as antiviral, also for Biochemical Research, clinically for the treatment of the diseases such as acute and chronic pharyngitis, lichen planus, verruca plana.N,O-Diacetylmuramidase is to adopt biotechnology to clone, extract and produce, and it is a kind of natural enzyme, and the additive of safe green, without resistance.This enzyme is extensively present in human multiple tissue, also containing this enzyme, wherein abundant with egg white content in the body fluid such as the egg white of birds and poultry, mammiferous tear, saliva, blood plasma, urine, milk and microorganism.From Ovum Gallus domesticus album, extracting separated N,O-Diacetylmuramidase is the single peptide chain consisting of 18 kinds of 129 amino-acid residues.It is rich in basic aminoacids, has 4 pairs of disulfide linkage to maintain enzyme configuration, is a kind of basic protein, and its N end is Methionin, and C end is leucine.Can decompose the gram positive organisms such as micrococcus lysodeikticus, bacillus megaterium, sarcina flava.N,O-Diacetylmuramidase is very stable protein, has stronger heat impedance.Egg white lysozyme is C type, is the most heat-resisting known enzyme.
Summary of the invention
The object of this invention is to provide a kind of method of preparing N,O-Diacetylmuramidase with egg white, N,O-Diacetylmuramidase is the abundantest with egg white content, and leaching process is simple, is applicable to scale operation.
To achieve these goals, technical scheme of the present invention is achieved in that the method for preparing N,O-Diacetylmuramidase with egg white, its operation steps is as follows: under room temperature, Ovum Gallus domesticus album is diluted to original 1.5-2 doubly with sodium chloride solution, with 2mol/L hydrochloric acid, regulate pH value 4-5.5, stir 45 minutes, the centrifugal removal precipitation of rotating speed and impurity with 6000r/min, collect supernatant liquor, the hollow fiber uf membrane system ultrafiltration that is 11000Da with retaining molecular weight by supernatant liquor, in gained filtrate, add ammonium sulfate solids to filtrate fractionation precipitation, supernatant liquor inclines, the spraying of gained precipitation is dry.
Further, the quality of ammonium sulfate solids accounts for 3% of filtrate volume.
Further, drying tower inlet temperature 110-160 ℃, temperature out 50-75 ℃.
The present invention has following beneficial effect: N,O-Diacetylmuramidase is the abundantest with egg white content, and leaching process is simple, and industrial production cost is low, is applicable to scale operation.
Embodiment
The present embodiment is prepared the method for N,O-Diacetylmuramidase with egg white, its operation steps is as follows: under room temperature, Ovum Gallus domesticus album is diluted to original 1.5-2 doubly with sodium chloride solution, with 2mol/L hydrochloric acid, regulate pH value 4-5.5, stir 45 minutes, the centrifugal removal precipitation of rotating speed and impurity with 6000r/min, collect supernatant liquor, the hollow fiber uf membrane system ultrafiltration that is 11000Da with retaining molecular weight by supernatant liquor, in gained filtrate, add ammonium sulfate solids to filtrate fractionation precipitation, the quality of ammonium sulfate solids accounts for 3% of filtrate volume, supernatant liquor inclines, the spraying of gained precipitation is dry, drying tower inlet temperature 110-160 ℃, temperature out 50-75 ℃.
Finally explanation is, above embodiment is only unrestricted in order to technical scheme of the present invention to be described, other modifications that those of ordinary skills make technical scheme of the present invention or be equal to replacement, only otherwise depart from the spirit and scope of technical solution of the present invention, all should be encompassed in the middle of claim scope of the present invention.
Claims (3)
1. with egg white, prepare the method for N,O-Diacetylmuramidase, it is characterized in that, its operation steps is as follows: under room temperature, Ovum Gallus domesticus album is diluted to original 1.5-2 doubly with sodium chloride solution, with 2mol/L hydrochloric acid, regulate pH value 4-5.5, stir 45 minutes, the centrifugal removal precipitation of rotating speed and impurity with 6000r/min, collect supernatant liquor, the hollow fiber uf membrane system ultrafiltration that is 11000Da with retaining molecular weight by supernatant liquor, in gained filtrate, add ammonium sulfate solids to filtrate fractionation precipitation, the supernatant liquor that inclines, the spraying of gained precipitation is dry.
2. the method for preparing N,O-Diacetylmuramidase with egg white according to claim 1, is characterized in that, the quality of ammonium sulfate solids accounts for 3% of filtrate volume.
3. the method for preparing N,O-Diacetylmuramidase with egg white according to claim 1, is characterized in that, drying tower inlet temperature 110-160 ℃, temperature out 50-75 ℃.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410269341.9A CN104004731A (en) | 2014-06-17 | 2014-06-17 | Method for preparing lysozyme by egg white |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410269341.9A CN104004731A (en) | 2014-06-17 | 2014-06-17 | Method for preparing lysozyme by egg white |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN104004731A true CN104004731A (en) | 2014-08-27 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN201410269341.9A Pending CN104004731A (en) | 2014-06-17 | 2014-06-17 | Method for preparing lysozyme by egg white |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105602918A (en) * | 2016-03-22 | 2016-05-25 | 烟台大学 | Method for extracting lysozyme from egg white or whole egg juice |
| CN106497900A (en) * | 2016-09-24 | 2017-03-15 | 合肥信达膜科技有限公司 | A kind of membrane treatment process for extracting lysozyme from egg white |
| CN107475221A (en) * | 2017-09-19 | 2017-12-15 | 青岛农业大学 | A kind of new lysozyme formulation and preparation method thereof |
Citations (7)
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|---|---|---|---|---|
| CN1285402A (en) * | 2000-09-18 | 2001-02-28 | 中国科学技术大学 | Process for separation of lysozyme |
| CN1464054A (en) * | 2002-06-03 | 2003-12-31 | 王峰 | Process for extracting lysozyme from eggs and bird eggs |
| CN1492041A (en) * | 2003-09-12 | 2004-04-28 | 山东农业大学 | Simple extracting method for high activity egg white lysozyme |
| CN101768577A (en) * | 2010-03-01 | 2010-07-07 | 德州天康食品有限公司 | Extraction of lysozyme in egg white and comprehensive utilization of egg white protein |
| CN102676475A (en) * | 2012-05-14 | 2012-09-19 | 石家庄华牧牧业有限责任公司 | Method for extracting muramidase from egg white |
| CN103160482A (en) * | 2013-03-14 | 2013-06-19 | 江苏大学 | Method for preparing egg white lysozyme and active protein by adopting coseparation |
| CN103555691A (en) * | 2013-11-08 | 2014-02-05 | 江南大学 | Method for extracting lysozyme from egg white |
-
2014
- 2014-06-17 CN CN201410269341.9A patent/CN104004731A/en active Pending
Patent Citations (7)
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|---|---|---|---|---|
| CN1285402A (en) * | 2000-09-18 | 2001-02-28 | 中国科学技术大学 | Process for separation of lysozyme |
| CN1464054A (en) * | 2002-06-03 | 2003-12-31 | 王峰 | Process for extracting lysozyme from eggs and bird eggs |
| CN1492041A (en) * | 2003-09-12 | 2004-04-28 | 山东农业大学 | Simple extracting method for high activity egg white lysozyme |
| CN101768577A (en) * | 2010-03-01 | 2010-07-07 | 德州天康食品有限公司 | Extraction of lysozyme in egg white and comprehensive utilization of egg white protein |
| CN102676475A (en) * | 2012-05-14 | 2012-09-19 | 石家庄华牧牧业有限责任公司 | Method for extracting muramidase from egg white |
| CN103160482A (en) * | 2013-03-14 | 2013-06-19 | 江苏大学 | Method for preparing egg white lysozyme and active protein by adopting coseparation |
| CN103555691A (en) * | 2013-11-08 | 2014-02-05 | 江南大学 | Method for extracting lysozyme from egg white |
Non-Patent Citations (2)
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| 金茜等: "从蛋清浓厚蛋白中提取溶菌酶技术的研究", 《农产品加工.学刊》, no. 298, 30 November 2012 (2012-11-30), pages 30 - 33 * |
| 黄建安等: "溶菌酶制备技术研究进展", 《江苏食品与发酵》, no. 109, 30 June 2002 (2002-06-30), pages 20 - 23 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105602918A (en) * | 2016-03-22 | 2016-05-25 | 烟台大学 | Method for extracting lysozyme from egg white or whole egg juice |
| CN106497900A (en) * | 2016-09-24 | 2017-03-15 | 合肥信达膜科技有限公司 | A kind of membrane treatment process for extracting lysozyme from egg white |
| CN107475221A (en) * | 2017-09-19 | 2017-12-15 | 青岛农业大学 | A kind of new lysozyme formulation and preparation method thereof |
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Application publication date: 20140827 |