Studies On Extraction, Purificati On And Structure As Well As Bio-Acticvities Of Coprinus Comatus Fruiting Polysaccharides

Coprinus comatus is a kind of edible and medicinal fungi. It is rich in polysaccharides. The polysaccharides with biological activity are widely used as an important resources for food, medicine industries. In this paper, modern isolation and analysis methods were used to study on the extraction, purification,characteristics and bioactivities of Coprinus comatus polysaccharides, systematically. The research offers some scientific basis for comprehensive utilization of Coprinus comatus.Crude polysaccharide was extracted by bioling water from fruiting bodies of Coprinus comatus. The best extraction and purification technology of Coprinus comatus polysaccharide were optimized with the orthogonal design. The result showed that the optimum condition for extraction process as follow: the amount of water added for the first time was 12 times that of dry-material ,the temperature was 95℃, and the total time of extraction was 1.5 h.The crde polysaccharide was fractionated and purification by ultrafiltration,anion-exchange chromatography and gel filtration chromatiography. Finally, four homogeneous polysaccharides (CBW-1,CBW-2,CBS-12,CBS-13) were obtained and homogeneous substance CCPS-3 were get by pre-process method, all were identified by HPLC.Analyzed by High Performance Anion Exchange Chromatography (HPAEC), we get the information that CBW-1 contained fucose, galactose, glucose and mannose in the ratio 1:3.81:0.08:0.09, with a molecular mass of 1.76×10~4 Da by HPLC; CBW-2, CBS-12 and CBS-13 contained fucose, galactose and glucose; each in the ratio of 1:4.12:0.12, 1:3.45:0.33, 1:3.81:0.42; molecular mass as follow: 1.76×10~4 Da,1.14×10~4 Da, 1.74~10~4 Da,1.17×10~4 Da.Homogenous substance CCPS-3 have 19.25 percent of glucose, it's molecular mass is 404. Adopt Sugar analysis, Methylation analysis and NMR to analysis the homogeneous polysaccharides, then we know the structure characterization of CBS-12 and CBS-13.①character analysis of CBS-12Results of Methylation analysis showed that: fucosc were terminal residues, galactose residues linked as 1,6, 1,2,6, branches attached to galactose residues. NMR analysis further showed that the CBS-12 consists pentasaccharide repeating units with (1,6)-linked a-D-galactopyranosyl residues as main backbone and substituted by a-L-fucopyranosyl residue. The structure as follow:②character analysis of CBS-13Results of Methylation analysis and NMR showed that CBS-13 has the same structure to the CBS-12, however, CBS-13 contains minor glucose as terminal resides, it's structure as follow:In the immunocompetence tests, CCPS effect on the proliferation of lymphocytes is the most obvious; CB,CC,CD have lightly effects on lymphocytes, meanwhile, CB is the top one; CB,CC,CD across anion-exchange chromatography get CBW-1,CBS-1,CBS-2,CCW-1,CCS-1,CCS-2,CDS-1,CDS-2 and all showed obvious effects on lymphocytes, meanwhile, fraction eluted by salt effects were stronger than that by water; CBW-1,CBW-2,CBS-12,CBS-13 as homogeneous polysaccharides have minor effects on lymphocytes, however, homogeneous subsetance CCPS-3 showed higher effects on lymphocytes than four homogeneous polysaccharides.