| Polysaccharide,also called polysaccharides,consists of more than 10 monosaccharides,which connected through glycosidic and widely exists in the organism.Polysaccharide plays an important role in the organism activities,such as anti-tumor,anti-virus,anti-oxidation and anti-bacterial activity.Pinus korongensis Pinon Shell Polysaccharides is extracted from the loose bulk shell waste of the Northeast of China,which has Anti-virus,anti-tumor,other biological activity and is widely paid attention.In this research,PPSP was firstly extracted by ultrasonic-assisted hot water extraction,then the PPSP-b which obtained after edulcoration,was isolated by ion-exchange chromatography and dextran gel chromatography and the physical and chemical properties of PPSP and its components were detected by UV spectrophotometer and chemical method.The extraction temperature of this study was 80 ?,extraction time was 40 min,the number of leaching was 2,and solid-liquid ratio was 1: 3.Result showed that the polysaccharide extraction rate was 2.37 ± 0.23% and the sugar content was 25.78%.PPSP was deproteinized by papain and Sevag method.The protein removal rate was 90.87% and the retention rate of polysaccharide was 76.41%.PPSP1,PPSP2 and PPSP3 were obtained through the PPSP-b DEAE-52 ion exchange chromatography separation.The PPSP2 and PPSP3 were further analyzed by Sephadex gel chromatography due to the content of PPSP1 is low and the PPSP2-1,PPSP2-2 and PPSP3-1,PPSP3-2 were obtaind,at last,the PPSP2-2 and PPSP3-1 were treat as the main research object because of the content of PPSP2-1 and PPSP3-2 was lower.The detection of physical and chemical properties showed that PPSP,PPSP2-2 and PPSP3-1 were mainly composed of polysaccharides.Then the primary structure of PPSP2-2 and PPSP3-1 in this study were measured.Result showed that there was three-helix conformation in PPSP2-2 molecule,while PPSP3-1 didn't exist.The result of the HPLC analysis showed that the composition and molar ratio of PPSP2-2 were Man: Rha: Glc A: Gal A: Glc: Gal: Xyl: Ara: Fuc =7.3: 12.1 :3.6 :11.8 :11.0: 20.2: 10.5: 18.4: 5.1,the monosaccharide composition and molar ratio of PPSP3-1 were;Man: Rha: Glc A: Gal A: Glc: Gal: Xyl: Ara: Fuc=4.6: 5.6: 2.9: 44.9: 23.8: 9.1: 2.5: 5.2: 1.3.The molecular weight distribution of PPSP2-2 and PPSP3-1 were relatively uniform,and the molecular weight are 1.89×103 Da and 2.84 ×103Da;The result of the infrared spectroscopy showed that PPSP2-2 and PPSP3-1 are acidic polysaccharides,which contained the pyranose.Finally,the effect of PPSP on the antioxidant activity of mice were examined,21 consecutive days of different doses PPSP was treated to mice by intragastric administration,the superoxide dismutase in serum and liver SOD,CAT,GSH-Px and the content of MDA the content of antioxidant index were detected.The results showed that the antioxidation of mice were increased,and showed a dose effect.Compared with the control group,high dose of PPSP group(400mg/(kg·D))could significantly increase SOD,CAT and GSH-Px activity of serum and the liver of mice(P<0.01),and significantly decreased the content of MDA(P<0.01);middle dose of PPSP group(200mg/(kg·D)significantly increased(P<0.01)SOD activity in liver,and significantly improve(P<0.05)the GSH-Px activity in serum of mice,and significantly decreased the MDA content in serum and liver of mice(P<0.05).In summary,four kinds of different polysaccharide components were obtained through the separation and purification of PPSP in this study,in which PPSP2-2 and PPSP3-1were the main components.The PPSP2-2 was mainly consisted of Gal,Ara,Rha,Gal A,Glc,Xyl monosaccharide and the molecular weight of that was 1.89×103 Da;the PPSP3-1 was mainly composed of Gal A,Glc,Gal and other monosaccharides,the molecular weight of that is 2.84 ×103Da.PPSP2-2 and PPSP3-1 are acidic polysaccharidestthat contain pyranose by infrared detection;PPSP has antioxidation biological activity in vivo,and which shows a dose effect. |
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