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In this thesis, mainly through bio-fermentation technology culture Phellinus pini ( Phellinus pini ) , to determine the optimal culture medium and fermentation conditions ; polysaccharide extracted from the mycelium of alkaline extraction method , purified alkaline extraction and water-soluble polysaccharide PEP and chemical structure of the polysaccharide and immune activity . Medium composition and fermentation conditions Phellinus pini mycelium growth and exopolysaccharide . The results showed that the the liquid fermentation optimum medium for 4% of white sugar , yeast extract 0.4 % , KH2PO4 0.3 % , MgSO4 0.1 % , VB1 , VB20.01g / L ; liquid fermentation , the optimal conditions for the incubation temperature 26 ° C , installed the fluid volume 250mL/500mL shaking speed 130r/min , initial pH of 6, 15% of the inoculum size , incubation time 6d . The mycelium biomass amounted to 5.27 g / L , up to 0.42 g / L extracellular polysaccharide production . Phellinus pini obtained by the fermentation of mycelium, the mycelium after water extraction extracted by 0.5 M NaOH at room temperature , hydrochloric acid and alcohol precipitation , conventional drying too alkaline extraction of water-soluble polysaccharide CPEP . The CPEP using freeze - thaw classification , the joint off enzymes and Sevage law protein and Sepharose CL - 6B column chromatography preparation purified to obtain a water-soluble polysaccharide PEP . Phenol - sulfuric acid method to measure the total sugar content of 92.12% . Sepharose CL - 6B column chromatography and HPLC analysis , PEP homogeneous polysaccharide , a molecular weight of about 16kDa . Partial acid hydrolysis , periodate oxidation , Smith degradation , methyl and IR, GC, GC-MS method for the analysis of the structure of the PEP . The consolidated results of several methods , suggesting that the the PEP structure of the main body portion is constituted by 1,2 - Man , the average every five residues on the two branches , the pivot connecting the side chain was separated in Man 6-O , 1,6 - Gal and 1,4 - Man a branched , terminal residues of Glc and Gal . Lymphocyte transformation in vitro experiments show that PEP can significantly promote the proliferation of lymphocytes in mice .
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