Study On Isolation, Purification, Sturvture And Antioxidant Activity Of Polysacchairides From Different Parts Of Pinus Koraiensis

In this study, this experiment to obtain polysaccharide from cone, bark and needleof Pinus Koraiensis using the method of deproteinization and depigmentation, also, wehave study physical and chemical properties and antioxidant capacity of the three kindsof polysaccharides respectively, concluded that antioxidant capacity of thepolysaccharide from cones is the strongest. Homogeneous polysaccharide ofPKP-D70-2-a and PKP-D70-2-b obtained by separating and purifying. Using combinedmethod of instruments and chemical to analysis chemical structure of PKP-D70-2-a andPKP-D70-2-b, also evaluated their antioxidant capacity. And finally to explore therelationship between structure and function, it will become waste into treasure, andgive their potential value.Determination of chemical components are mainly contains polysaccharides anduronic acid, but the content of proteins and polyphenol are small, less than4.5%of thetotal using the method of phenol sulfuric acid, m-hydroxyldiphenyl method, coomassiebrilliant blue method, folin phenol.By infrared spectroscopy, three kinds ofpolysaccharides are polysaccharides having characteristic absorption peaks. the threepolysaccharide contain seven kinds of polysaccharides, the D-ribose, L-rhamnose,L-arabinose, D-xylose, D-mannose, D-glucose, D-galactose by acid hydrolysiscompletely. The polysaccharide from cone hasve good antioxidant capacity, whichscavenge hydroxyl radicals, ABTS radicals and reducing power of EC50values were6.54,5.06,9.15mg/mL.Two homogeneous polysaccharide PKP-D70-2-a and PKP-D70-2-b obtained use themethod of graded alcohol precipitation with ethanol, deproteinized, depigmentation,DEAE-52and Sephadex G-100column chromatography. The threee polysaccharidesmain contain polysaccharides and uronic acid, and a little sulfuric acid group,phosphoric acid group, benzoyl group. At the same time,using ultraviolet spectrum,infrared spectrum, nuclear magnetic resonance spectroscopy, complete acid hydrolysis,periodate oxidation-smith degradation, partial acid hydrolysis, Congo red test, ?elimination reaction to analysis chemical structure of homogeneous components. PKP-D70-2-a contains five kinds of monosaccharides, which are L-rhamnose,L-arabinose, D-mannose, D-glucose, D-galactose, and PKP-D70-2-b contains four kindsof monosaccharides, which does not contain the L-rhamnose. The backbone and sidechains of PKP-D70-2-a are composed of five kinds of monosaccharides, the backboneand side chain of PKP-D70-2-b contain four monosaccharides, mainly connect with1?2,1?2,6,1?4,1?4,6. Branched chain sugar content accounted for55.49%inPKP-D70-2-a,53.80%in PKP-D70-2-b. In both glycoproteins contain O-sugar bond.PKP-D70-2-a in aqueous solution shows the main free coil conformation, andPKP-D70-2-b shows the main helix conformation. PKP-D70-2-a glycosidic bondconfiguration are3,4)-a-Galp-(1-2)??4)-?-D-GalpA-(1?and4,6)-?-GlcpNAc-(1-4),PKP-D70-2-b glycosidic bond configuration are ?-D-Glcp-(1???-D-Gal link a-D-Gal-6sulfate??4)-?-D-Glc-(1???4)-?-D-GalpA6Me-(?? ?-D-Glc(1?).Through the antioxidant capacity evaluation, PKP-D70-2-b free radical scavengingability of PKP-D70-2-b was better than PKP-D70-2-a, but the reducing power wasweaker than that of PKP-D70-2-a. Also through the preliminary structure-activityrelationship studies, the relative molecular mass, replace the antioxidant capacity of thebase type, monosaccharide composition and configuration have great influence on theantioxidant activity of PKP-D70-2-a and PKP-D70-2-b.