Extraction, Isolation, Structure Identification And Biological Activity Of Polysaccharides From Yunzhi Fruiting Body

Corilus versicolor polysaccharide is one of the important bioactive substance,widely used in health food and medicine.The Flash Extraction process of C.versicolor polysaccharide was optimized by Box-Benhnken response surface design.The condition of flash extraction was 2.88 min of extraction time,16667.25 r/min of speed,1:40 of solid-liquid ratio.Under the optimal conditions C.versicolor Polysaccharide yield was 4.94%.Polysaccharide extract graded ethanol precipitation to give three fractions was named CVPS30,CVPS60,CVPS80 obtained by gradient alcohol precipitation.Polysaccharide content of CVPS30 was up to 59.14%,so we chose CVPS30 to separate and purify.Crude polysaccharide CVPS30 was purified by DEAE-Sepharose Fast Flow ion chromatography and Sephacryl S series(S-200 and S-500)gel column chromatography,then obtained CVPS30-2aI.After analyzing by high performance liquid chromatography and near-infrared spectroscopy,it was proved to be a neutral polysaccharide with 1.937×106 Da of molecular weight.Studies have shown that,the monosaccharide composition of CVPS30-2aI was Dglucose detected by gas-mass spectrometry after Acetylation.Methylation analysis detected the glucose residues of CVPS30-2a I were mainly four kinds of connections,respectively,4-Glcp,4,6-Glcp,3,6-Glcp and T-Glcp,and the molar ratio was 3.33:2.25:1:2.82.Vitro anti-oxidation experiments showed that CVPS30,CVPS60,CVPS80 had good anti-oxidant activity,and the concentration was higher,the anti-oxidant activity was better.They had strong scavenging activity to the ABTS radical and hydroxyl radical and relatively weak scavenging activity to the DPPH radical.MTT assay was applied to detect the proliferation of spleen lymphocytes.The results showed that the four kinds of Corilus versicolor polysaccharide had direct effect on spleen lymphocyte proliferation,and also had synergistic proliferation effect on spleen lymphocyte induced by ConA with concentration effect in 50-200?g/mL.Wherein CVPS80 had the strongest direct proliferation and synergy proliferation effect,and the direct proliferation and synergy proliferation activity of CVPS30-2aI was significantly higher than unpurified CVPS30.