The Applanatum Polysaccharide Isolated And Purified, Structure Identification And Its Biological Activity

Polysaccharide were extracted with water by orthogoral design and isolated by ethnol step by step,DEAE cellulose ion exchange chromatography and sephadex G10 gel chromatography. The homogeneous polysaccharide were obtained.The contents of neutral sacchride and uronic acid of Gaps were determined by spectrophotometry .Homogeneity and molecular weight were determined by HPLC combined with GPC with IR detector;Composition of monosacchride by GC and GC-MS;Chemical Structure were analyzed by methods of chemical and spectroscopy.The contents of neutral sacchride was 45% as well as 11% of uronic,mainly composed of glucose,galose,mannose,rhamnose,xylose and arabinose.Gaps contained three fractions that 80% of the total less than 6000D.Five homogeneous polysaccharides(Gaps3TⅠ,Gaps3TⅡ,GapsⅠ,GapsⅡ,GapsⅢ)were isolated and purified. the structures of the repeating unit of two polysaccharides ( Gaps3TⅠ,GapsⅢ) were established.The composition of monosacchride and molecular weight of other three polysaccharides were determined.The Gaps were seperated with ethnal step by step,fraction of three times ethna(lGaps3T) is homogeneous on both molecular weight and polarity. The Mw was estimated to be 4800D by HPLC. The Gaps3T was further seperated by dialysis,inner part(Gaps 3TⅠ)and inter part (Gaps 3TⅡ) were obtained. Homogeneity and molecular weight were determined by HPLC with topMw 5300D,4300D respectly. The contents of Gaps3TⅠwas 80.8%.Gaps3TⅠmainly composed of glucose and manose as well as galactose,xylose,arabinose,ribose,fucnose in a molar ratio 37.68:4.47:1:0.63:0.98:0.82:0.97.On the basis of partial acid hydrolysis,periodateoxidation and Smith degradation,methylation analysis,IR,UV,NMR studies,the structure of the repeating unit of the polysaccharide was established as :α-D-Manp(1→6)-β-D-Manp(1→3)- [β-D-GLCp(1→3)-β-D-GLCp(1→]9Three homogeneity polysaccharides were obtained by DEAE and sephadex G10 gel chromatography,named as GapsⅠ,GapsⅡ,GapsⅢ. They are homogeneous on both molecular weight and polarity by HPLC.The TopMw was estimated respectly to be 6621D,5535 D,3210D by HPLC.The contents of GapsIII was 94.3%,composed of glucose,galactose,mannose,xylose,arabinose ,ribose fucose,rhamnose in a molar ratio 15.30: 1:0.77:0.14:0.14:0.86:0.16:0.90.Compositional analysis,methylation analysis and IR,UV,NMR studies revealed the presence of the following reapting unit in the polysaccharide.The contents of Gaps3TⅡwas 86.9%,mainly composed of glucose,galose ,mannose,xylose, arabinose,ribose,fucose,rhamnose in a molar ratio 31.91:1:3.22:0.41:0.20:0.41:0.46:0.36. The contents of GapsⅠand GapsⅡwere 90.8%,93.0%.GapsⅠwas mainly composed of glucose,galose,mannose,arabinose,ribose,fucose in a molar ratio 13.73:1:1.01:0.91:0.43:0.31:0.46; GapsⅡis mainly composed of glucose as well as galactose,manose,xylose,arabinose,ribose,fucose,rhamnose in a molar ratio 13.97:1:1.47:1.18:1.48:1.24:0.29:0.15.Gaps had the inhibitory action on the gastric mucosal induced by acetic acid,the stress,the medicine (indomethacin) and the pylorus ligation,its anti-ulcer function increased along with the dosage strengthens. Sets up the Gaps also to be possible obviously to increase stomach mucous membrane blood stream quantity of gastric ulcer induced by the acetic acid in rats and to enhance the stomach mucous membrane barrier function. To ties up the pylorus gastric ulcer,sets up the Gaps to affect not obviously to the gastric juice volume,but had the inhibitory action to the gastric juice and pepsin activeness,indicated that its may reduce the aggressive factor to the stomach wall's corrosion and the injury.A colorimetric tetrazolinm Gaps possessed the obvious antiflammatory and antinociceptive effects.Gaps had the inhibitory action both on the the foot swelling of rats induced by carrageenin and on auricle edema of mice induced by xylene, and inhibitory action on analgesic action of mice.MTT method was used for the determination of anti-tumor activity eraluation,the effect was significant,Gaps3T? and GapsⅢshowed antitumor activity evidently.Five homogeneous polysaccharides were seperated and identified ,and the structure of the repeating unit of Gaps3TⅠand GapsⅢwere established using many kinds of genealogy methods for the first time. The anti-tumor activeness of Gaps3TⅠand GapsⅢthat top molecular weight are 5300D and 3520D respectively were studied for the first time.The experiments proved that Gaps had the strong anti-ulcer activeness for the first time. The research of Gaps and 5 kind of homogeneous polysaccharides had laid the foundation for the new medicine's research and development, had provided the scientific basis for the Ganoderma applanatum further research.