Fecal Coprinus Mycelium Polysaccharide Separation And Purification, Structural Analysis And Inhibition Of S-180 Tumor

This paper studied on the fractionation, purification, physicochemical property, structure characteriztion and anti-tumor activity of polysaccharides from Coprinus sterquilinus Fr.Crude polysaccharide was extracted by hot water and precipated by alcohol. And then, protein was removed by using Sevag method, enzymatic, trichloroacetic acid method and enzymatic combination of Sevag method, to select the best method of them. Polysaccharide was purified by DEAE-Cellulose-32 and Sepharose-6B gel chromatography and identified by Sepharose-6B gel chromatography and Rotation method to obtain homogeneous polysaccharide components. On this basis, physicochemical property and structural characteriztion of homogeneous polysaccharide CPS-1a were analyzed by gel column chromatography, thin layer chromatography, GC, iodine reaction, UV, IR,13CNMR, 'HNMR, partial acid hydrolysis and other methods. At last, the inhabit effects of crude polysaccharide of Coprinus sterquilinus Fr. was reseached combination of pharmacological test for mouse S-180 tumor cells.The results showed that enzymatic combination of Sevag method had the best effects among four methods which Sevag method, enzymatic, trichloroacetic acid method and enzymatic combination of Sevag method, and content of protein was 1.90%, content of polysaccharide was 4.53%, removal rate of protein was 70.95%, loss rate of polysaccharide was 6.60%. CPS-1 and CPS-2 were isolated from CPS using DEAE-Cellulose-32, CPS-1a, CPS-1b and CPS-1c was isolated from CPS-1 using Sepharose-6B.The results showed that CPS-1a was homogeneous polysaccharide and molecular weight was 54.5×104Da by Sepharose-6B gel chromatography and Rotation method; GC analysis showed it was composed of xylose, glucose, galactose and rhamnose which the mole ratio was 1.38:12.85:4.81:1.00.The structure of CPS-1a was analyzed by gel column chromatography, thin layer chromatography, GC, iodine reaction, UV, IR,13CNMR,1HNMR, partial acid hydrolysis and other methods. The results showed that CPS-1a not contained a continuousα-(1â†'4) Glc, proteins, nucleic acids,βconfiguration and glucuronic acid. CPS-la was a kind of neutral polysaccharide andαconfiguration of D-glucopyranose. Its main chain was composed of Glc, terminal of branched-chain or main chain was composed of Gal, Xyl, and Rha, edges of branched-chain or main chain was composed of Glc.Experimental of anti-tumor results showed that the S-180 mouse were feeded with crude polysaccharide from Coprinus sterquilinus Fr., the index which spleen and thymus of mouse not had obvious effect.