|
Inonotus obliquus (Inonotus obliquus (Fr.) Pilat), is a rare large-scale consumption of medicinal fungi, in Russia more than a hundred years of application history, can treat a variety of diseases. Humfeld H mushrooms submerged fermentation in conducting research by fermentation culture mushroom mycelium, the researchers have been using this method for a variety of fungus fermentation. Inonotus obliquus at home and abroad submerged fermentation less Inonotus obliquus polysaccharide deep fermentation extraction and purification as well as its chemical properties are more scarce. In this paper, Inonotus obliquus strains deep fermentation. In order to get a lot of mycelium and polysaccharides, using the single factor experiment and response surface analysis of the combination of mycelium and polysaccharide optimized medium, said dry weight DNS method, the phenol-sulfuric acid method, vanillin - ice acetic acid method of the mycelial content, reducing sugar, total sugar, of triterpenoids fermentation product content in the fermentation broth; the fungal polysaccharides extraction and separation steps, get the two obliquus polysaccharide IO2b,, IO3a. Concluded as follows: (1) Inonotus obliquus mycelia optimal culture conditions: corn saccharification liquid 2.76%, 1.9% soluble starch, 0.75% yeast extract, peptone 0.95%, the KH 2 PO 4 0.2%, the MgSO 4 0.05%; the mycelium biomass accumulation time 6D; optimum culture temperature of 28 ° C; medium The initial pH 5; determine the best shaker speed 130r/min; inoculation amount was 15%; liquid volume of 250 mL each flask 70mL medium. (2) the Inonotus obliquus extracellular polysaccharide optimal culture conditions: corn saccharification liquid 2.1%, 2.23% soluble starch, 0.51% yeast extract, peptone 0.41%, the KH 2 PO 4 0.2%, MGSO 4 0.05%; determine 130r accumulation of extracellular polysaccharide 7d; 28 ° C for optimal incubation temperature; medium initial pH 6; / min for the best speed; 15% of the inoculum size; liquid volume of 250mL flask 70mL medium. Inonotus obliquus (3) submerged culture mycelia and liquid separation, fermentation broth was concentrated alcohol precipitation, removal of protein, dialysis, drying steps to get the Inonotus obliquus polysaccharide IO. DEAE-52 and Sephadex G-200 of the fractionation to get the two obliquus the polysaccharide IO2b and IO3a. (4) the use of paper chromatography to determine the of Inonotus obliquus monosaccharide composition of glucose, galactose, mannose and arabinose. The infrared spectrum shows characteristic absorption peak of the polysaccharide HPLC measured molecular weight of IO2b is a 1.64 × 10 5 , molecular weight of IO3a as 8.26 × 10 5 . The chemical reaction that is a polysaccharide substance, and does not contain starch molecules having long side chains and more branching.
|