Heparitin sulfate from rat tissue and production thereof

A technology of heparan sulfate and tissue, applied in the field of heparan sulfate and its preparation, can solve the problems of limited research and application of heparan sulfate, few types of heparan sulfate, no heparan sulfate, etc., and achieves rich variety and specific composition. , structure-specific effects

Inactive Publication Date: 2006-08-16
OCEAN UNIV OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the types of heparan sulfate obtained are few, the structure lacks diversity, and the anticoagulant is low, which greatly limits the research and application of heparan sulfate
So far, there are no reports on heparan sulfate derived from mouse tissue

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0009] After 100 g of rat liver tissue was minced and homogenized, it was degreased with chloroform and methanol at volume ratios of 1:1, 1:1, and 1:2 in sequence. Grind after drying; suspend the obtained rat liver powder in 100mL of 0.1mol / L Tris-hydrochloric acid and water buffer solution, adjust the pH of the hydrolyzate to 9.0 with 0.05mol / L sodium carbonate aqueous solution, add 2.7 unit chain Mycoprotease was hydrolyzed at 50°C for 40h, centrifuged at 4°C to get the clear liquid; the clear liquid was hydrolyzed at 45°C for 8h in 200mL 0.2mol / L sodium hydroxide aqueous solution containing 50mmol / L sodium borohydride, and added 5% (100% by weight Concentration, the same below) perchloric acid aqueous solution, centrifuge at 4°C to take the clear liquid, and dialyze with distilled water for 1 day (MWCO, 3500); Place it under the hood for 3 hours, collect the precipitate by centrifugation, wash the obtained precipitate once with 0.1% cetylpyridinium chloride aqueous solution...

Embodiment 2

[0012] After 50 grams of fresh rat spleen tissue was minced and homogenized, it was degreased with chloroform and methanol at a volume ratio of 10 times the volume of 2:1, 1:1, and 1:2. Grind after drying; suspend the obtained rat spleen powder in 50 mL of 0.1 mol / L Tris-acetic acid and water buffer solution, adjust the pH of the hydrolyzate to 8.6 with 0.05 mol / L sodium carbonate aqueous solution, and add 1.5 units of base Protease was hydrolyzed at 40°C for 20 hours, centrifuged at 10°C to get the clear liquid; the clear liquid was hydrolyzed in 100mL0.2mol / L sodium hydroxide aqueous solution containing 50mM sodium borohydride aqueous solution for 16h at 40°C; add 15% perchloric acid aqueous solution , centrifuge at 10°C to get the clear liquid, dialyze with distilled water for 3 days (MWCO, 3500); add 25mL of 10% cetylpyridinium chloride aqueous solution to the obtained dialyzate, place it at 10°C for 6h, centrifuge to collect the precipitate, and the obtained precipitate R...

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PUM

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Abstract

Heparan sulfate originating from rat tissue and its production are disclosed. The procedures is carried out by homogenizing rat tissue, degreasing, hydrolyzing by protease, hydrolyzing by alkali, deproteinizing, centrifugalizing, taking out purified liquor, dialyzing, depositing dialyzate by cetylpyridinium chloride, hydrolyzing by RNase and chondroitinase ABC, and separating by ionic exchanging column. It has special raw material, better product anticoagulant and antiradiation performances, and various Heparan sulfate. It can be used for research on medicinal molecular model structure and protein interaction. I

Description

technical field [0001] The invention relates to a heparan sulfate derived from rat tissue and a preparation method thereof. Background technique [0002] Heparan sulfate is a glycosaminoglycan present in various animal cell matrices and cell membranes, and is one of the most complex biological macromolecules in nature. Heparan sulfate sugar is composed of hexuronic acid (glucuronic acid or iduronic acid) and acetylglucosamine disaccharide units repeatedly, because the amino and hydroxyl groups in the sugar chain are substituted by sulfate groups to varying degrees, resulting in its structural diversity. There are 32 combinations of disaccharide units, and heparan sulfate composed of 10 monosaccharides may have millions of structural sequences. Thus, heparan sulfate becomes the biomolecule with the highest density of biological information. [0003] The chemical composition, structure and activity of heparan sulfate obtained from different animal and tissue sources and by ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08B37/10
Inventor 毛文君
Owner OCEAN UNIV OF CHINA
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