Study On Separation, Purification And Structure Of Polysaccharide From Abalone Harslet

Polysaccharide was extracted from abalone harslet(AHP) by alkalinity proteinase. By a further separation and purification, we obtained a homogeneous polysaccharide(AHP-12), and the molecular weight, composition, primary structure of this polysaccharide were determined.After the treatment with pepsin and Sevage method, pronase and Sevage method were applied to deproteinate the abalone harslet polysaccharide. The reliquus protein was deleted by dialysising and freeze thawing. The abalone harslet polysaccharide was obtained by cryodesiccate, and its protein content was only16.93%.Then the crude AHP was purified by Sephadex G-100gel and DEAE-cellulose52chromatography, finally we got the purified AHP-12fraction. It was a homogeneous polysaccharide demonstrated by HPLC, agarose gel electrophoresis and UV spectrum. Its molecular weight was about300kDa. Monosaccharide composition analysis indicated that AHP-12was composed of rhamnose, fucose, galactose, mannose and xylose, the molar ratio was Rha:Fuc:Gal:Man:Xyl=l1.6:11.6:9.6:1.3:1. It contained13.07%sulfate and4.98%aminohexose, aminogalactose was3.54%among the aminohexose.The structure of the AHP-12was investigated by using various spectroscopic techniques and chemical methods. The results of partial hydrolysis, Periodate oxidation, Smith degradation and G.C-M.S dealing with methylation showed:(1) The main chain was composed of fucose and galactose, and the proportion was1:1;(2) In the main chain, Gal was existed in the form of (1â†'3) inoxidable glycosidic bond and Fuc was existed in the form of (1â†'4) oxidable glycosidic bond;(3) Gal(1â†'3) and Fuc(1â†'4) glycosidic bond linked alternately;(4) Xyl(1â†'2,3) and Man(1â†'4) glycosidic bond should existed on the side chains, the main chain was branched by Xyl(1â†'2,3) glycosidic bond;(5) Aminoglucose and xylose were existed in the form of (1â†') glycosidic bond, so they should located at the terminate. According to the above news, the possible repeated unit of the AHP-12was: Man(1â†'4),...