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Purification method for extracting glycoprotein from rhopilema esculentum

A purification method and glycoprotein technology, applied in the field of extraction and purification of glycoproteins from jellyfish, can solve the problems of large molecular weight of glycoproteins, achieve the effects of reducing desalination procedures, improving efficiency, and rapid concentration

Inactive Publication Date: 2012-05-02
HENAN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to solve the problem of the large molecular weight of glycoproteins obtained from jellyfish in the prior art, the present invention provides a method for extracting and purifying glycoproteins from jellyfish, obtaining jellyfish glycoproteins with lower molecular weight and uniform components, and separating and purifying them

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0027] Weigh 1000g of fresh minced jellyfish, add 4000ml of NaCl solution with a concentration of 0.3mol / L, place it in a constant temperature shaker at 10°C for 10h, then take it out and send it to a centrifuge, Centrifuge at 4000r / min for 20min, take the supernatant; the supernatant is then concentrated to 1 / 20 of the original volume with a hollow cellulose membrane, and added with a mass concentration of 95% to the concentrated supernatant while stirring. ethanol, when the mass concentration of ethanol in the solution reaches 75%, stop adding ethanol, let it stand for 2h, and then centrifuge for 15min under the condition of 10000r / min at a centrifugal speed, get the precipitate and freeze-dry to obtain 0.58g crude sugar protein dry powder; repeat In the above steps, a sufficient amount of glycoprotein is obtained.

Embodiment 2

[0029] Weigh 1000g of fresh minced jellyfish, add 4000ml of NaCl solution with a concentration of 0.5mol / L, place it in a constant temperature shaker at 10°C for extraction for 10h, then take it out and send it to a centrifuge. Centrifuge at 4000r / min for 20min, take the supernatant; the supernatant is then concentrated to 1 / 20 of the original volume with a hollow cellulose membrane, and added with a mass concentration of 95% to the concentrated supernatant while stirring. ethanol, when the mass concentration of ethanol in the solution reaches 75%, stop adding ethanol, let it stand for 2h, then centrifuge 15min under the condition of 10000r / min at the centrifugal speed, get the precipitate and freeze-dry to obtain 1.23g crude sugar protein dry powder; repeat In the above steps, a sufficient amount of glycoprotein is obtained.

Embodiment 3

[0031] Weigh 1000g of fresh minced jellyfish, add 4000ml of NaCl solution with a concentration of 0.7mol / L, place it in a constant temperature shaker at 10°C for extraction for 10h, then take it out and send it to a centrifuge. Centrifuge at 4000r / min for 20min, take the supernatant; the supernatant is then concentrated to 1 / 20 of the original volume with a hollow cellulose membrane, and added with a mass concentration of 95% to the concentrated supernatant while stirring. ethanol, when the mass concentration of ethanol in the solution reaches 75%, stop adding ethanol, let stand for 2h, then centrifuge 15min under the condition of centrifugal speed of 10000r / min, get the precipitate and freeze-dry to obtain 1.20g crude sugar protein dry powder; repeat In the above steps, a sufficient amount of glycoprotein is obtained.

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Abstract

The invention relates to a purification method for extracting glycoprotein from rhopilema esculentum, comprising the following steps: adding a sodium chloride solution in a tissue smashing liquid obtained after fresh rhopilema esculentum is smashed, and extracting; then, concentrating by using a hollow cellulose film; adding ethanol after the concentration; freeze-drying a generated sediment to obtain crude glycoprotein; purifying the obtained crude glycoprotein again through DEAE (Diethylaminoethyl) cellulose ion exchange column chromatography and gel column chromatography; and collecting eluent and freeze-drying to obtain purified glycoprotein of rhopilema esculentum. In the invention, the glycoprotein is extracted from a fresh rhopilema esculentum raw material by using a sodium chloride extract, thus, novel glycoprotein different from the past glycoprotein extracted from the rhopilema esculentum is obtained; the hollow cellulose film is used for concentrating, thus, compared with the traditional rotary evaporative concentration method, the purification method is capable of realizing low-temperature, mass and rapid concentration and has the effect of removing macromolecular impurities; and ion exchange and gel column chromatography are adopted in the following process, and the eluent is bi-distilling water, thus, the following desalting process is omitted, and the extracting and purifying efficiencies are increased.

Description

technical field [0001] The invention relates to a method for extracting and purifying glycoprotein, in particular to a method for extracting and purifying glycoprotein in jellyfish. Background technique [0002] The research on marine bioactive substances is a research hotspot in the development and utilization of marine resources. The diversity and specificity of marine organisms provide humans with a wide variety of natural products with novel structures and specific activities. So far, humans have isolated more than 10,000 compounds from various marine organisms, nearly one-half of which have various biological activities. The ocean is a huge treasure house for human beings to obtain medicines, functional foods, biological materials, and food. [0003] 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 natu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K1/36C07K1/18C07K1/16
Inventor 任国艳康怀彬郭金英梁旺春刘志龙樊金玲陈秀金李道敏
Owner HENAN UNIV OF SCI & TECH
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