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Method for extracting and purifying dermatan sulfate in angler fish skin

A technology of dermatan sulfate and a purification method, which is applied in the field of biopharmaceuticals, can solve the problems that fish skin cannot be used, and achieve a reasonable effect of the method

Inactive Publication Date: 2008-01-09
ZHEJIANG GONGSHANG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The purpose of the present invention is to provide a method for extracting and purifying high-purity dermatan sulfate from anglerfish skin to obtain stable quality dermatan sulfate, while solving the practical problem that fish skin cannot be used in processing anglerfish, and aiming at the current sources of most dermatan sulfate Develop raw materials with more development potential based on the current situation of pigs and cattle

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] (1), the raw material is 10g of dried anglerfish skin. Add 0.125g of papain, 0.125g of trypsin, and 300mL of distilled water, extract in a water bath at 70-75°C for 17-19 hours, cool and centrifuge to obtain 225mL of supernatant.

[0026] (2) Add 4.500 g of sodium hydroxide and 0.851 g of sodium borohydride to the supernatant, stir to dissolve the solid, and measure NaOH and NaPH 4 The final concentrations in the supernatant were 0.5 mol / L and 0.1 mol / L respectively, treated in a water bath at 45-50°C for 24 hours, centrifuged, and the pH of the supernatant was adjusted to neutral with acetic acid, with a total of 203 mL of liquid.

[0027] (3) Add 68mL of protein-removing solution (chloroform: n-butanol=2:1 (v:v)) to the liquid, shake it fully for 20-25 minutes, put it in a separatory funnel and let it stand for stratification. Then discard the lower liquid.

[0028] (4), supernatant repeat step (3) operation 4 times. 68 mL of supernatant was obtained.

[0029] (5)...

Embodiment 2

[0031] (1) The raw material is 10g of dried anglerfish skin. Add 0.100 g of papain, 0.250 g of trypsin, and 300 mL of distilled water, extract in a water bath at 51-56°C for 13-15 hours, cool and then centrifuge to obtain 228 mL of supernatant.

[0032] (2) Add 4.560g of sodium hydroxide and 0.862g of sodium borohydride to the supernatant, stir to dissolve the solid, treat it in a water bath at 30-35°C for 30 hours, centrifuge, adjust the pH of the supernatant to neutral with acetic acid, and the liquid 210mL.

[0033] (3) Add 70mL of protein-removing solution (chloroform: n-butanol=3:1 (v:v)) to the liquid, shake it well for 15-20 minutes, put it in a separatory funnel and let it stand for stratification. The lower liquid was discarded.

[0034] (4) Repeat step (3) for the supernatant 4 times. 70 mL of supernatant was obtained.

[0035] (5) Add 140mL ethanol to the supernatant, let it stand at 4°C for one day, collect the precipitate, dissolve it with 50mL distilled water...

Embodiment 3

[0037] (1) The raw material is 10g of dried anglerfish skin. Add 0.050 g of papain, 0.100 g of trypsin, and 260 mL of distilled water, extract in a water bath at 65° C. for 17 hours, and centrifuge after cooling to obtain 200 mL of supernatant.

[0038] (2) Add 67mL of protein-removing solution (chloroform: n-butanol = 4.5:1 (v:v)) to the liquid, shake it thoroughly for 30 minutes, put it in a separatory funnel to stand for layers, and discard after layers underlying liquid.

[0039] (3) Repeat step (2) for the supernatant 4 times. 67 mL of supernatant was obtained.

[0040] (4) Add 134mL ethanol to the supernatant, let it stand at 4°C for one day, collect the precipitate, dissolve it with 50mL distilled water, centrifuge to get the supernatant, put it in a dialysis bag with a molecular weight of 14000, and dialyze it against running tap water for two days, and then dialyze it for one day with distilled water. After dialysis, the liquid was freeze-dried to obtain 0.147 g of...

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Abstract

It is concerned with extraction and purification of sulfuric acid skin-lotion from waste skins of angler fish production. It is carried out by: extracting with compounded enzyme of pancreatin and papain, adding NaOH into extracting liquid to cut off glycopeptide bond while protecting glycosidic bond with NaPH4, removing protein by savage process, depositing sulfur acid skin lotion in alcohol, dissolving in water and centrifuging dissoluble substances, dialyzing to remove ions and micro-molecular substances, and frozen drying high purity sulfur acid skin lotion with ratio of sulfate radical to carboxylic radical = 1.07. The product is strong antithrombotic, with less side effect of hemorrhage, has disinfecting activity, and can be used in pharmaceutical field.

Description

technical field [0001] The invention belongs to the technical field of biopharmaceuticals, and relates to the preparation of biomedicine derived from animal bodies, in particular to a method for extracting and purifying dermatan sulfate from anglerfish skin. Background technique [0002] Dermatan sulfate (DS) is a mucopolysaccharide compound and is the most important glycosaminoglycan in animal skin. Its molecules are mainly composed of double sugar chains composed of N-acetylgalactosamine 4-sulfuric acid and L-iduraldehyde In addition, there are D-glucuronic acid double sugar chains, N-acetylgalactosamine 6-sulfate and D-glucuronic acid double sugar chains. It has strong anticoagulant activity, as well as regulating blood lipids, anti-inflammation, and anti-atherosclerosis. As a drug, it has the characteristics of strong antithrombotic effect and long half-life. Compared with heparin, its outstanding advantage is that it has no bleeding side effects and is well tolerated b...

Claims

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Application Information

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IPC IPC(8): C12P19/04A61P7/02A61P29/00
Inventor 张虹廖文娟张燕平
Owner ZHEJIANG GONGSHANG UNIVERSITY
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