Preparation method for micro-molecular glycopeptide of jellyfish

A small-molecule sugar and jellyfish technology, which is applied in the field of preparation of jellyfish small-molecule glycopeptides, can solve the problems of unfavorable glycopeptide structure research and large molecular weight of glycopeptides.

Inactive Publication Date: 2012-06-27
HENAN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In order to solve the problem that the glycopeptide extracted by the prior art has a relatively large molecular weight and is unfavorable for further research on the structure of the glycopeptide, the present invention provides a method for preparing a small-molecule glycopeptide from jellyfish

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] One, the preparation of jellyfish glycoprotein, its operation is as follows:

[0020] 1) Grind the fresh jellyfish with a tissue masher, weigh 200g of the tissue mash solution, add 800ml of NaCl solution with a concentration of 0.5mol / L, mix well, and extract in a constant temperature shaker at 10°C for 10h. Then take it out and send it to a centrifuge, centrifuge for 20min under the condition of a centrifugal speed of 4000r / min, get the supernatant, and set aside;

[0021] 2) Concentrate the supernatant in step 1 to 1 / 20 of the original volume with a hollow cellulose membrane, add ethanol with a mass concentration of 95% to the concentrated supernatant while stirring, when the mass concentration of ethanol in the solution is When it reaches 75%, stop adding ethanol, let it stand for 2 hours, centrifuge for 15 minutes under the condition of centrifugal speed of 10000r / min, take the precipitate and freeze-dry it to be crude glycoprotein, and set aside;

[0022] 3) Use D...

Embodiment 2

[0029] One, the preparation method of jellyfish glycoprotein is as embodiment 1;

[0030] 2. The preparation of jellyfish small molecule glycopeptides, the operation steps are as follows: Take 1 g of glycoprotein separated and purified by ion exchange and gel column chromatography and dissolve it in distilled water to make a glycoprotein solution with a mass concentration of 0.1 mg / mL. Add compound protease to the protein solution for enzymolysis, the amount of compound protease added is 0.5% of the mass of the glycoprotein solution, enzymolysis at pH 8.0 and temperature 65°C for 2 hours to obtain glycopeptide solution, using ultrafiltration method, The obtained glycopeptide solution was graded, ultrafiltered with 3000Da, and the filtrate was concentrated and freeze-dried to obtain jellyfish small molecule glycopeptides. It was determined that 90% of the glycopeptides had molecular weights less than 3000Da.

Embodiment 3

[0032] One, the preparation method of jellyfish glycoprotein is as embodiment 1;

[0033] 2. The preparation of jellyfish small molecule glycopeptides, the operation steps are as follows: Take 1 g of glycoprotein separated and purified by ion exchange and gel column chromatography and dissolve it in distilled water to make a glycoprotein solution with a mass concentration of 0.1 mg / mL. Add compound protease to the protein solution for enzymolysis, the amount of compound protease added is 0.7% of the mass of the glycoprotein solution, enzymolysis is carried out at pH 8.0 and temperature 65°C for 2 hours to obtain a glycopeptide solution, and ultrafiltration is used. The obtained glycopeptide solution was graded, ultrafiltered with 3000Da, and the filtrate was concentrated and freeze-dried to obtain jellyfish small molecule glycopeptides. It was determined that 90% of the glycopeptides had molecular weights less than 3000Da.

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Abstract

The invention provides a preparation method for micro-molecular glycopeptide of jellyfish. The method comprises the following steps: adding a sodium chloride solution into tissue trituration liquid obtained after trituration of fresh jellyfish for extraction, carrying out condensation by using a hollow cellulose film, adding ethanol for deposition so as to obtain crude glycoprotein, then purifying the crude glycoprotein by using DEAE cellulose ion exchange column chromatography and gel column chromatography, collecting and freeze-drying obtained eluate so as to obtain purified jellyfish glycoprotein, carrying out enzymatic hydrolysis on the purified jellyfish glycoprotein by using composite protease so as to obtain a glycopeptide solution and freeze-drying a filtrate obtained after ultrafiltration of the glycopeptide solution so as to obtain the micro-molecular glycopeptide of jellyfish. Compared to traditional enzyme methods in which glycopeptide is directly obtained in an organism, the invention has the following advantages: influence of other ingredients in a resultant on the activity of enzyme is avoided, the enzyme directly and accurately acts on glycoprotein, the activity of the enzyme is brought into full play, and the micro-molecular glycopeptide is obtained. The micro-molecular glycopeptide obtained in the invention can be used for mass spectrometry, so the micro-molecular glycopeptide has important significance to structural analysis of glycoprotein.

Description

technical field [0001] The invention relates to a preparation method of glycopeptide, in particular to a preparation method of jellyfish small molecule glycopeptide. Background technique [0002] Jellyfish is a large edible jellyfish with the same origin of medicine and food, which has high nutritional value and medicinal value. Jellyfish is low in sugar, low in fat and high in protein, so it is a kind of natural health food. Modern medical research shows that jellyfish has the functions of lowering blood pressure, lowering blood fat, preventing atherosclerosis, and beauty and skin care. Due to the characteristics of high water content and easy autolysis of jellyfish, the early research on jellyfish mainly focused on toxins. However, with the continuous advancement of science and technology and modern instrumental analysis methods, there are more and more studies on active ingredients in jellyfish, such as toxins, polysaccharides, glycosaminoglycans, collagen, collagen pol...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/06
Inventor 任国艳康怀彬郭金英刘志龙梁旺春陈秀金崔国庭樊金玲
Owner HENAN UNIV OF SCI & TECH
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