Method for preparing sialoglycopeptide (SGP) by large-scale separation and purification

A sialic sugar, large-scale technology, applied in the field of biochemistry, can solve the problems of high cost, low cost, long cycle, etc., and achieve the effects of low cost, lower overall cost, and mild elution conditions

Active Publication Date: 2019-05-31
NORTHWEST UNIV(CN)
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] Aiming at the shortcomings of existing sialoglycopeptide separation techniques, such as many steps, long cycle time, and high cost, the purpose of the present invention is to provide a short cycle, low cost, and high efficiency separation and purification preparation of sialoglycopeptide (SGP) Methods

Method used

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  • Method for preparing sialoglycopeptide (SGP) by large-scale separation and purification
  • Method for preparing sialoglycopeptide (SGP) by large-scale separation and purification
  • Method for preparing sialoglycopeptide (SGP) by large-scale separation and purification

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Effect test

Embodiment 1

[0043] Specific steps are as follows:

[0044] (1) First take 50 fresh unfertilized egg yolks (1L), add an equal volume of deionized water to dilute, and stir evenly with a rotor. In order to achieve the purpose of protein denaturation, add 10 times the volume of phenol aqueous solution (water:phenol=9:1) to the above egg yolk solution, keep stirring for 2 hours, then add 2L of double distilled water and mix well, then centrifuge (4500g, 30min), The supernatant was concentrated to 50mL under reduced pressure with a rotary evaporator, transferred to a 250mL round bottom flask, and the flask was frozen at -80°C for 1 hour, and then the frozen sample was dried in a vacuum freeze dryer.

[0045] (2) Take 150 mg of medical absorbent cotton and stuff it evenly into a 1 mL pipette tip, and compact it slightly. Wash the cotton column with 10 mL of double distilled water, and then wash the cotton column with 10 mL of 100% acetonitrile. Dry the cleaned cotton column, and place it in an...

Embodiment 2

[0057] Specific steps are as follows:

[0058] (1) First take 50 fresh unfertilized egg yolks (1L), add an equal volume of deionized water to dilute, and stir evenly with a rotor. In order to achieve the purpose of protein denaturation, add 10 times the volume of phenol aqueous solution (water:phenol=9:1) to the above egg yolk solution, keep stirring for 2 hours, then add 2L of double distilled water and mix well, then centrifuge (4500g, 30min), The supernatant was concentrated to 50mL under reduced pressure with a rotary evaporator, transferred to a 250mL round bottom flask, and the flask was frozen at -80°C for 1 hour, and then the frozen sample was dried in a vacuum freeze dryer.

[0059] (2) Take 150 mg of medical absorbent cotton and stuff it evenly into a 1 mL pipette tip, and compact it slightly. Wash the cotton column with 10 mL of double distilled water, and then wash the cotton column with 10 mL of 100% acetonitrile. Dry the cleaned cotton column, and place it in an...

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Abstract

The invention discloses a method for preparing sialoglycopeptide (SGP) by large-scale separation and purification. The method comprises (1) taking fresh unfertilized egg yolk, treating with phenol, centrifuging to obtain a supernatant and lyophilizing to obtain coarse sialoglycopeptide; (2) filling a cotton hydrophilic chromatographic column with medical degreasing cotton; (3) adding a solution ofthe crude sialoglycopeptide to the cotton hydrophilic chromatographic column; (4) conducting gradient elution on the cotton hydrophilic chromatographic column with acetonitrile to remove salt and impurities and conducting elution and lyophilization on the glycopeptide with water to obtain pure sialoglycopeptide. According to the method, the cumbersome steps of Sephadex G-50 separation, Sephadex G-25 desalination, ion exchange chromatography purification and the like are omitted, the reaction conditions are mild, the experiment cost is reduced, the purification cycle is shortened, the efficiency is improved, and the purity of the sialoglycopeptide is 95% or above according to the purity detection. The method is not only suitable for enrichment of laboratory glycopeptide but also used for industrial production.

Description

technical field [0001] The invention relates to a method for separation and purification of glycopeptide compounds, especially focuses on the separation and purification of sialic acid glycopeptides, and belongs to the technical field of biochemistry. Background technique [0002] Sialic acid glycopeptides (Sialylglycopeptides, SGP) is a glycopeptide extracted from fresh egg yolk with natural complex N-glycopeptides, composed of eleven monosaccharides and six amino acids, and its non-reducing ends are usually two A sialic acid, also known as disialoglycopeptide (its structure is shown below). [0003] [0004] SGP has many important biological activities. For example, it can be used as an antagonist to inhibit the combination of bacteria and viruses with host cells. In terms of antibacterial, it can inhibit the combination of Salmonella and other bacteria with human intestinal cells and mouse tissue cells. The sialylated α-(2→6)N-acetyllactosamine residue in SGP can also...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K9/00C07K1/22C07K1/14
CPCY02A50/30
Inventor 王仲孚韩健利黄琳娟
Owner NORTHWEST UNIV(CN)
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