Optimization And Establishment Of Extracting Polysaccharide Technologies From Acathopanax Senticosus And Pholiota Nameko

As an immune curing remedy, Polysaccharide had become a new researching spot whether at home or at abroad. But only few compound polysaccharides are developed, such as "Wu Xian Ji" in recent years. For preparing products of compound polysaccharides, the pure polysaccharides must be made. However, the technologies of extraction and purification of polysaccharide either from plants or mushroom exist many problems to be solved by research. This study tried to establish the new technologies of extraction and purification of polysaccharide. Results were as following.Using Acathopanax senticosus and pholiota nameko as experimental material, the factors, such as the ratio of sample and water, the time of extraction, the temperature of extraction and the times of extraction on the extraction of the water soluble polysaccharides was investigated by the methods of single factor tests and orthogonal design test. The results of extracting polysaccharide from Acathopanax senticosus as follow: adding distilled water at 18:1 ratio volume, water bath at 90℃for 150 minutes, then extracting polysaccharide of Acathopanax senticosus by alcohol at 4: 1 ratio by volume, suitable pH 7, at -20℃for 36 hours. The best conditions of extracting polysaccharide from pholiota nameko was adding 40 times of water volume as much that of the sample, extracting 90 minutes at 90-100℃, one times of extraction times.To study decolorization and deprotein of the extraction technology of polysaccharide, many reagents were compared, such as 860021 resin,DM2 resin,330 resin,D941 resin,D202R resin,active carbon and H₂O₂. the best method of decolorization and deprotein was D941 resin. Using D941 resin, the ratios of decolorization and deproteinization and polysaccharide reservation were 84.39%, 85.00%, 83.38%, respectively. The validation tests by Lentinan indicated the ratios of decolorization and deproteinization and polysaccharide reservation were 87.829%, 92.34%, and 68.89%, respectively. This suggested D941 resin methods was feasible in the extraction technology of polysaccharide. To study the separation and refinement technology of polysaccharide from Acathopanax senticosus, DEAE Sepharose F. F column were applied to separate polysaccharide from the extract of Acathopanax senticosus. Two kinds of polysaccharide components were obtained and Sephadex-G200 was used to determine the purity of refined polysaccharide.The optimum strain of Pholiota nameko used for submerged fermentation was screened and the strain of C3-3 Linsheng was the best among the thirteen strains of Pholiota nameko preserved in our laboratory. The best carbon and nitrogen source of the media suitable for the strain were evaluated. Fructose was the best carbon among the carbon source, such as fructose, dissolubility starch, corn flour, glucose, maltose and mannose. The wheat bran juice was better nitrogen sources than yeast powder, yeast extract, soybean powder, ammonium sulphate and ammonium nitrate. The best culture medium includes dissolubility starch 15.0 g/L, wheat bran juice 15.0 g/L, MgSO₄0.1 g/L, KH₂PO₄1.5g/L.