Mactra veneriformis glycosaminoglycan extraction method

Abstract

The invention provides a Mactra veneriformis glycosaminoglycan extraction method, which is characterized by comprising the following steps of: weighing the soft part of Mactra veneriformis, adding water to prepare into a homogenate with the ratio of water to material being 3:1, adding subtilisin and trypsin into the homogenate for enzymatic hydrolysis while the ratio of bacillus subtilis neutral proteinase to trypsin is 4:3, the addition of compound enzyme is 1% and the hydrolysis time is 3-5h, boiling and centrifuging the enzymolysis liquid to obtain a clear liquid, adding active carbon for decolouring, followed by vacuum filtration, filtering with the assistance of diatomite, carrying out alcohol precipitation to obtain crude glycosaminoglycan with the yield rate being more than 1.3%, preparing 3% of a sugar solution, centrifuging to remove acidic and neutral proteins, followed by dialysis of a supernatant and alcohol precipitation, preparing 1% of a sugar solution, adding 5% of CTAB, centrifuging, dissolving the precipitate into a KCI solution to obtain a dissociation liquid, carrying out alcohol precipitation, followed by dialysis and alcohol precipitation to obtain high-purity glycosaminoglycan. The method provided by the invention has advantages of mild reaction condition and high yield rate and purity. Lots of proteins can be recovered while extracting glycosaminoglycan. The extracted substance has important effects of removing free radial, resisting oxidation and resisting coagulation. The yield rate of crude glycosaminoglycan reaches over 1.3% and the purity of glycosaminoglycan reaches more than 95%.

Description

technical field [0001] The invention relates to a method for extracting glycosaminoglycan, in particular to a method for extracting and preparing clam glycosaminoglycan. Background technique [0002] At present, there are three main extraction methods of glycosaminoglycans: the first non-degradation method refers to the method of extracting glycosaminoglycans from animal and plant tissues with water or saline solution; the second alkaline extraction method is based on the glycopeptide bonds in proteoglycans. For alkali instability, the sugar and protein are separated by alkaline extraction; the third enzymatic degradation method, protease degrades the protein part of the proteoglycan to separate the sugar chain. Among the three methods, enzymatic degradation is the most widely used method. There have been some reports on the extraction of glycosaminoglycans from shellfish, but the composition, content and functional activity of glycosaminoglycans in different shellfish are d...

AI Technical Summary

Problems solved by technology

Among the three methods, enzymatic degradation is the most widely used method. There have been some reports on the extraction of glycosaminoglycans from shellfish, but the composition, content and functional activity of glycosaminoglycans in different shellfish are different. From the point of view of the extraction and preparation of shellfish glycosaminoglycans, there are generally defects of low yield and purity, so the present invention can make up for the above defects