Extraction, Purification And Structure Analysis Of Polysaccharides From Agaricus Blazei Murill

In this dissertation, the extraction and purification of polysaccharides from Agaricus Blazei Murill fruiting body were systematic studied. A high molecular weight polysaccharide mono-fraction was purified, and some physicochemical characters and structural properties of this fraction were researched.The results are mainly as following:1. Some mono-factors in extracting the fruit bodies of Agaricus Blazei Murill with water were firstly studied, then through orthogonal test, the most rational technologic factors were obtained, with manufacture considerated, the finally rational technology condition were: 100 grams Agaricus Blazei Murill powder was soaked with 3000ml water (weight of material :weight of water=l:30), firstly treated with microwave for 10 minutes ,then extracted at temperature of 120C for 3h; Vacuum concentrated to proper volume, then it was precipitated with 3 times volume of 95% ethanol for 6h at room temperature, and the yield of polysaccharide was up to 9.12%, which was higher than that mentioned in literature.2. When in period of purification for above crude polysaccharides, Sevag method, enzyme decomposition with Sevag method, trichloroacetic acid(TCA) sedimentation, ultrafiltration , anion and cation exchange resin chromatography were studied to remove the protein in the polysaccharides.Based on experimental results and general consideration, it was suitable to adopt using anion and cation exchange resin in series chromatography once. After that, 92.36% protein of crude polysaccharides could be removed, the recovery rate of polysaccharide was 82.85%, and the ratio value of polysaccharides content to protein content was up to 19.6.Then after dialysis and freeze dryness, light yellow powder was produced, we called it as fine Agaricus Blazei Murill polysaccharides. In this powder, polysaccharide and protein content was 89.27% and 4.55%.Several gel column chromatography were used to separate fine polysaccharide. Firstly, higher molecular and lower weight sections were gained by using Sephadex G-200 gel chromatograph. Then a polysaccharide fraction containing 1.5% protein was gained by Sephacryl S-400 chromatographing to the higher molecular. The purity of this fraction was proved by HPGPC and its molecular was more than 2 X 106 daltonand was named A.B.M.F-VHMW. The portion of A.B.M.F-VHMW was 28.96% of fine polysaccharides. And the low molecular weight was chromatographed by Sephadex G-100 gel filtration, and three fraction were gained , the molecular weight of them was 7.2 X 104(38.2%), 4.8 X104 (16.1%), 1.3 X 104(45.7%) separately.3.Some physicochemical characters and structural properties of A.B.M.F-VHMW were researched. The optical rotation degree of A.B.M.F-VHMW was [a]D25- +62.5?, and it was soluble in water, insoluble in ethanol, aether, acetone. After completely hydrolyzation by 2.0mol/L H2SO4 and analysed by HPLC it was confirmed that A.B.M.F-VHMW was made up of glucose only. Then through IR, HC NMR, the results showed that the main polysaccharide chain was a - (1 6 ) D-Glc. and 1.5% combined protein was contained.The creative points are as folio wings:1 .Micro-wave treated and high pressure were Combined originally and the yield was greatly improved when in extraction,.2.For the first time, anion and cation exchange in series chromatography resin were adopted to remove the protein, and the ideal results were gained.3.An A.B.M.F-VHMW polysaccharide fraction which molecular was more than was purified by several gel filtration, and some physicochemical characters and structural properties of it were researched, the results showed that the main polysaccharide chain was a - (1 ?) D-Glc contained with 1.5% combined protein, and it was not reported in domestic and overseas literature.