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Lamb fourth stomach glycoprotein extraction, separation and purification method and application thereof

A technology for separation and purification of glycoproteins, used in the development and application of natural source glycoproteins, to achieve the effects of short time consumption, high extraction rate and mild conditions

Active Publication Date: 2021-09-28
XINJIANG BIOCHEM PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Glycoproteins in the gastrointestinal tract cover the surface of the gastric mucosa, which has a protective effect on the gastric injury site, increases the value of gastric parietal cells, and promotes mucosal repair; however, there is no research report on the preparation and function of lamb's fourth gastric glycoprotein

Method used

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  • Lamb fourth stomach glycoprotein extraction, separation and purification method and application thereof
  • Lamb fourth stomach glycoprotein extraction, separation and purification method and application thereof
  • Lamb fourth stomach glycoprotein extraction, separation and purification method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Extraction of Lamb's Fourth Stomach Glycoprotein:

[0041] a. Take 30g of the fourth stomach of fresh lamb, wash the contents of the stomach cavity with tap water, peel off the fat and connective tissue, freeze at -80°C for 3 hours, cut it into pieces, crush it with liquid nitrogen, and freeze-dry it to obtain the fourth stomach of lamb powder;

[0042] b. The lamb fourth stomach powder obtained in step a was magnetically stirred, and the petroleum ether was defatted twice to obtain the lamb fourth stomach defatted powder, which was stored in a freezer at a temperature of -20°C;

[0043] c. Add the degreased lamb fourth stomach powder obtained in step b into 2L of absolute ethanol and magnetically stir to remove alcohol-soluble small molecule impurities, repeat twice, filter under reduced pressure, and air-dry naturally to obtain degreased and decolorized lamb fourth stomach powder;

[0044] d. Using a 0.5% sodium chloride solution with a solid-to-liquid ratio of 1:30 ...

Embodiment 2

[0055] Extraction of Lamb's Fourth Stomach Glycoprotein:

[0056] a. Take 30g of the fourth stomach of fresh lamb, wash the contents of the stomach cavity with tap water, peel off the fat and connective tissue, freeze at -80°C for 3 hours, cut it into pieces, crush it with liquid nitrogen, and freeze-dry it to obtain the fourth stomach of lamb powder;

[0057] b. The lamb fourth stomach powder obtained in step a was magnetically stirred, and the petroleum ether was defatted twice to obtain the lamb fourth stomach defatted powder, which was stored in a freezer at a temperature of -20°C;

[0058] c. Add 2L of absolute ethanol to the degreased lamb fourth stomach powder obtained in step b, magnetically stir to remove alcohol-soluble small molecule impurities, repeat twice, filter under reduced pressure, and air-dry naturally to obtain degreased and decolorized lamb fourth stomach powder;

[0059] d. Add the powder obtained in step c with 1.5% sodium chloride solution at a materi...

Embodiment 3

[0070] Extraction of Lamb's Fourth Stomach Glycoprotein:

[0071] a. Take 30g of the fourth stomach of fresh lamb, wash the contents of the stomach cavity with tap water, peel off the fat and connective tissue, freeze at -80°C for 3 hours, cut it into pieces, crush it with liquid nitrogen, and freeze-dry it to obtain the fourth stomach of lamb powder;

[0072] b. The lamb fourth stomach powder obtained in step a was magnetically stirred, and the petroleum ether was defatted twice to obtain the lamb fourth stomach defatted powder, which was stored in a freezer at a temperature of -20°C;

[0073] c. Add 2L of absolute ethanol to the degreased lamb fourth stomach powder obtained in step b, magnetically stir to remove alcohol-soluble small molecule impurities, repeat twice, filter under reduced pressure, and air-dry naturally to obtain degreased and decolorized lamb fourth stomach powder;

[0074]d. Add the powder obtained in step c to 3% sodium chloride solution at a material-to...

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Abstract

The invention relates to a lamb fourth stomach glycoprotein extraction, separation and purification method and application thereof. The lamb fourth stomach glycoprotein extraction, separation and purification method comprises the following steps that a pretreated fresh lamb fourth stomach is degreased, and ultrasonic-assisted extraction is carried out on a sodium chloride solution; centrifuging is carried out to take a supernatant; vacuum concentration and alcohol precipitation are carried out; free protein is removed with a Sevage reagent; freeze-drying is carried out to obtain crude glycoprotein powder; the glycoprotein is obtained through gel and anion exchange column chromatography separation and purification, the yield is 1.67%, the total sugar content is 70.24%, the protein content is 16.39%, and the uronic acid content is 4.12%; and purifying is carried out by HPLC to obtain the high-purity lamb fourth stomach glycoprotein, and the purity reaches 99.894%. In-vitro probiotic activity experiments prove that the lamb fourth stomach glycoprotein has the effect of promoting the growth of intestinal probiotics of bifidobacterium adolescentis, lactobacillus delbrueckii subspecies bulgaricus and enterococcus faecalis. The lamb fourth stomach glycoprotein extraction, separation and purification method has the advantages of simple process, convenience in operation, high extraction rate, high purity, stable property, no need of using a large amount of toxic solvents, low cost, easiness in popularization and application and the like.

Description

technical field [0001] The invention relates to the field of development and application of a natural source glycoprotein, in particular to a method for extracting, separating and purifying lamb fourth stomach glycoprotein and its application. Background technique [0002] Glycoproteins are macromolecular complexes formed by covalent linkages of oligosaccharides and polypeptide chains. Glycoproteins widely exist in animals, plants and certain microorganisms, and have biological activities such as enhancing immune regulation, inhibiting tumors, antioxidation, antibacterial, and anticoagulant. [0003] In recent years, the relationship between intestinal probiotics and overall human health has attracted more and more attention. Prebiotics are nutrients that can feed the intestinal microbiota and promote the growth of intestinal probiotics. Active substances such as secondary metabolites glycopeptides, short-chain fatty acids (SCFAs), lactic acid, butyric acid, and propionic ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K1/14C07K1/36C07K1/30C07K1/16C07K1/18C07K1/34A61K38/17A61P1/00A23L33/17
CPCC07K1/145C07K1/36C07K1/30C07K1/16C07K1/18C07K1/34A61P1/00A23L33/17A61K38/00A23V2002/00A23V2200/32A23V2250/204A23V2250/5438Y02P60/87
Inventor 阿布力米提·伊力阿吉艾克拜尔·艾萨赵新民阿米娜·艾尔肯
Owner XINJIANG BIOCHEM PHARMA CO LTD
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