Study On The Extraction, Purification And Antitumor Activity Of Polysaccharides From Sporisorium Reilianum By Liquid Fermentation

Sporisorium reilianum, also was named Sphacelotheca reiliana, commonly called smut fungus, belong to Basidiomycolina, Teliomycetes, Sporisorium, Ustilaginaceae, Ustilaginales. It is one of the most important pathogenic fungus of plant.But Sporisorium reilianum were natural health food and can provide abundant nutrition for the body, because they were rich in nutrition, such as protein, minerals, vitamins. It was reported that smut fungus exhibited activities such as anti-tumor, anti-inflammatory, hypoglycemic and strengthen haematogenous function. At present at home and abroad, the researches on Sporisorium reilianum were focused on the classification and pathogens to the plants. The use of Sporisorium reilianum on food was limited; some scholars'studies use them to product. Polysaccharide is a important active material, it accounted for 3.5% of the quality of the Sporisorium reilianum, and become the hot spot of the studies. But the amount of smut fungus in the nature makes the development and exploitation suffer confinement by the seasonal factors. One way to obtain polysaccharide is from fruiting bodes. Liquid fermentation is the main way to overcome this deficiency.The paper based on isolation and identification of Sporisorium reilianum, then gain mycelium polysaccharide by liquid fermentation. The polysaccharide was isolation, purification, and was studied on the bioactivity.1. The purpose of this chapter was to isolate and indentify Sporisorium reilianum from "Ji Liang 105A". In this study, pure strain was isolated by isolate single spore. The morphology of the isolated strain were observed and the ITS were sequenced. The results demonstrated that ITS amplified fragment length is 721bp. Through the comparison analysis of the sequences by the DNAstar software, found that the isolated strain was Sporisorium reilianum. The sequence was submitted to Genebank, get Accession number is HQ641393.2. Optimization of liquid fermentation medium and cultivating conditions of Sporisorium reilianum. The monofactorial test showed that soluble starch and soybean powder were the best carbon sources and nitrogen sources. Then, the addition of carbon sources, nitrogen sources and mineral salt were discussed by orthogonal test. The results showed the optimum fermentation medium composition was as follow: soluble starch 3%, soybean powder 0.3%, KH2PO4 0.3%, MgSO4 0.1%. The suitable cultivating conditions were as follow:initial pH of medium 5.0, culture temperature 28℃,100mL liquid medium in size 250mL shaking flask, inoculation quantity 8%, shaking frequency 130r/min, time 7d. The biomass of mycelian was 0.3895g/100mL. 3. Studies on the isolation, purification and the bioactivity of the polysaccharides from. Sporisorium reilianum by liquid fermentation. The suitable technique of extracting polysaccharides from Sporisorium reilianum mycelium is:temperature of extraction 90℃, solid-liquid ratio 1:60, extraction time 3h, times of extraction 3. Extraction ratio of polysaccharide is 8.51%. In order to obtain the purified polysaccharide, the effects of sevag and TCA for removing protein from the polysaccharides by comparing the percentage of the decomposed polysaccharide and the removed protein. The results choose sevag and 3 times. To study decolorization of the polysaccharide, many reagents were compared, such as active carbon, D315resin, D113resin and H2O2, the best method of decolorization was D113resin. Took out of salt and small molecular substance with dialyses, separated polysaccharides with Sephadex G-150 column chromatography. The molecular weight of the polysaccharide was 176906.4. Studies on the immunomodulating activities and acute toxicity experiment of Sporisorium reilianum polysaccharide. Bleeding tine and coagulation time were measured in mice by means of tail cutting and glass slide showed Sporisorium reilianum polysaccharide shorted bleeding tine and coagulation time. Inhibition effect on engorgement of mice induced by xylene. The results show that crude polysaccharide from Sporisorium reilianum polysaccharide obvious inhibition effect. The Sporisorium reilianum polysaccharide to sp20 tumor have good inhibitory effect, and had no toxic effect on biology organism. LD50>10g/kg·bw show that the polysaccharide from the mycelium of Sporisorium reilianum by liquid fermentation is in fact innocuous.