Study On Isolation, Purification And Initial Structure Of Water-soluble Polysaccharide Of Phellinus Igniarius

The extraction and isolation of Phellinus igniarius from mycelium were studied in this paper. Then process flow of purification of polysaccharide was established. And the structure of three water-soluble polysaccharide were detected.Hot water extraction method, microwave-assised extraction method and supersonic extraction method were optimized by using phenol sulfuric method and orthogonal experiment, in the standard of observing and studying the content polysaccharide of phellinus igniarius. By analysis of range, the optimized condition of hot water extraction method: extracting 3 hours at 90℃for three times, liquid/solid ratio being 50, the extraction rate of content polysaccharide was 2.10%; the optimized condition of microwave-assised extraction method: processing 15 min for three times, liquid/solid ratio was 50, the extraction rate was 4.18%; the optimized condition of supersonic extraction method: processing 30 min at 60℃for twice, liquid/solid ratio being 60, frequency being 60Hz, the extraction rate was 3.02%. The Sevag, TCA and TCA-E were used to remove the protein of polysaccharide from mycelium and fermentation. The results of three methods showed that the rate of protein-removed were 82.4%, 71.3%, 79.6% and the rate of polysaccharide-lost were 51.5%, 8.9%, 9.8%. The TCA-E was the most suitable method in the range of laboratory by extracting 15min, using 10% of TCA and using 1.2% of enzyme.Two fractions designated PI1, PI2 from mycelium and one fractions designated PI3 from fermentation were isolated by gradient ethanol precipitation, decolorization, dialysis. The yields of them were 13.5%, 12.4%, 15.9%. PI11, PI21, PI31 fractions were isolated by Sephadex G-100. The purities were 98.7%, 99.5%, 98.9%. The homogenicities of them were proofed by ultrafiltration and freeze thawing.All of the three fractions were masculine in Molish, showed special absorption peaks in IR. They did not content the glucosidic bond ofα(1→4), because of the negative in reaction of I. They soluted in water, especially in the hot water. But not in organic solvent, for example ether, acetone, ethanol. No absorption peaks at 260nm and 280nm shows that there were no nucleic acid and protein. The molecular weight of PI11, PI21 and PI31 were 65000, 30000 and 12000. The moleculars were made up by Xyl, Man, Fuc, Glc and Gal. The mole rate of PI11 was 5.2: 1: 10.2: 9.24: 30.1; PI21 was 0.08: 1: 4.6: 6.9: 14.1; PI31 was 2.3: 1: 6.4: 22.1: 19.8.The aim of the present research was to detecte a high-efficient and facility technology of extraction, isolation and purification water-soluble polysaccharide of Phellinus igniarius, and collected amont. The conclusion could provide some theory evidence for further explore on the principles of anticancer and others.