Study On Isolation, Purification, Structure And Bioactivities Of The Polysaccharides From Agaricus Blazel Murill

This paper studied on the isolation,fractionation,physicochemical properties, structural analysis and immunological activity of polysaccharides from Agaricus blazei.Murill(ABM) fruiting body.The relationship between the structure and activities of the polysaccharides was also discussed.Crude polysaccharide was extracted by hot water and precipated by alcohol.And then, was fractionated by anion-exchange and gel chromato graphy.Further,the structural features were determined by chemical methods such as periodate oxidation,Smith degradation,methylation and spectroscopy methods including infrared spectrum,GC-MS, 13C-NMR.Finally,the immunological activity of Agaricus blazei.Murill polysaccharide was assayed in vivo.The ABM was first cut into small pieces; followed by extraction with hot water. The crude polysaccharide was composed of Glc, Man and trace of Gal and Ara. No uronic acid The content of carbohydrate was 38.6%determined with phenol-sulfuric acid method. The content of protein was 35.4%measured with Bradford method and the iodine coloration method was used to determined the content of starch not have.By ethanol Preciptation,Polysaccharides from ABM were separated in two groups,ABM1 and ABM2,of yield 47.4% and 39.5%.After being deproteinated by Sevag method, the polysaccharides were fractionated by Sepharose CL-6B gel chromatography to obtain one fractions ABMH.ABMH was a homogeneous polysaccharide identified by high-performance liquid chromatography(HPLC)and its molecular weight was 7500 Da..Sugar analysis showed it was composed of Glc (41.5%), Gal (25.7%) and Man (32.5%).The structure of ABMH was determined by composition analysis,molecular weight analysis, periodate oxidation,Smith degradation,methylation,infrared spectrum and nuclear magnetic resonance(13NMR)analysis.The results showed that ABMH was consitituted major by (1→6)-D-Glc, (1→2,6)-D-Gal and (1→6)-D-Gal. Residues are substituted at 2-O of(1→2,6)-D-Ga,There is one branch every 7 sugar residues on the average.The bioassay showed that Agaricus blazei.Murill polysaccharide had potent stimulating effects on proliferation of both T and B lymphocytes in vivo.