The Study Of Structure And Bioactivity Of The Polysaccharide Extracted By Superalkali From The Mycelium Of Phellinus Pini

A study was made on effects of Medium composition and fermentation condition for the mycelial growth and the production of extracellular polysaccharide during fermentation of phellinus pini. The results showed the optimum medium compositions were edible sugar4%,yeast powder 0.4%,KH2PO4 0.4% ,MgSO4 0.1%,VB1,VB2 0.01g/L;the optimum cultural period was 6d at 26℃in 500 mL shake flasks at 130r/min.The best aeration ratio was 1 : 2(medium volume:flask volume).The inoculation volume was 15%.The hyphostroma biomass reached 5.27 g/L and the extracellular polysaccharide of phellinus pini reached 0.42g/L in the optimum culture.In addition , Extracellular polysaccharide of the content and the effect on the immunomodulatory activities of murine spenic lymphocytes in vitro were analyzed.A water-soluble polysaccharide isolated with 0.5M NaOH from the mycelium of Phellinus pini at room temperature.The suspension was neutralized with hydrochloric acid (1M) and filtered. The suspension containing water-soluble polysaccharide was dialyzed,concentrated,lyophilized and then dried,yielding the crude polysaccharide CPEP.PEP was obtained from CPEP by frozen-thawed,deproteinated with enzyme/Sevag method and Sepharose CL-6B column chromatography. The content of polysaccharide was 92.12%. The molecular weight of it was1.6×104 Da ,which measured by HPLC.The structure of PEP was elucidated by periodate oxidation,Smith degradation, methylation analysis and IR,GC,GC-MS. The results indicated that PEP consisted of a backbone composed of (1→2)-linked-Man and (1→2,6)-linked-Man residues in the ratio of 1:1.5.The side chains composed of (1→4)-linked–Man and (1→6)-linked-Gal residues in the ratio of 1:1.5 and The nonreduced end was Glc and Gal.Immunobilological activity assay showed that PEP could increase the ConA or LPS-induced lymphocytes proliferation in vitro.