Study On The Isolation,Purification, Structure, Antioxidation And Antibacteria Of Polysaccharide From Dictyophora Rubrovolvata

Dictyophora is the fungi of the phalloidin Section Dictyophora, and is a kind of delicious and precious food. There are12species around the world, and7species in China. Dictyophora rubrovolvata is one of the best, beautiful shape, nutrient-rich and delicious.Dictyophora rubrovolvata is the treasures in the Dictyophora, is a kind of basidiomycetes. And values as edible and medicinal provisions. Contains a variety of biological actives ingredients,such as amino acid, vitamin, inorganic salts, and polysaccharides, and has the function of anti-hypertensive, anti-hypoglycemic, anti-inflammatory, anti-cancer, anti-oxidation, immunological and so on. The polysaccharides is one of the major components with biological activities. Now, The study for polysaccharides of Dictyophora rubrovolvata is not very deeply and comprehensive. Therefore, in this work, choose Dictyophora rubrovolvata as raw material. Study on the isolation, purification, structural analysis, in vitro antioxidant and antibacterial activity of Dictyophora rubrovolvatan polysaccharides. The main conclusions of this article are as follows:(1) This is a experiment studies on the extraction process of polysaccharides of Dictyophora rubrovolvata used hot-water extraction method. On the base of the single-factor experiments, using response surface method to optimize the test design, fitted into a second-order polynomial equation by applying multiple regression analysis, and the regression model was thus predicted in code form: Y=0.113867+0.001551×X2+0.0011X3+0.005054xX4-0.016834×X12-0.01336×X22-0.015177×X32+0.002763×X3xX4-0.01697×X42.The optimum extraction conditions were as follow:extraction time3.51h, extraction temperature75.3℃, ratio of water to raw material31, and number of extraction2.Under the conditions,the extraction ratio of the polysaccharides is12.05%.(2) Crude polysaccharides of Dictyophora rubrovolvata was fractionated by DEAE-52anion exchange chromatography and purified by Sephadex G-100gel filtration chromatography to obtain three fractions, named DR1, DR2, DR3. The purity is identified by the specific rotation and HPLC. The result showed that they are all homogeneous.(3) Gas chromatography (GC) was used to dentermine the monosaccharide composition of DR1, DR2and DR3. The results showed that they are composed of glucose, xylose and galactose respectively. With the method of HPLC, the molecular weights of DR1, DR2and DR3are6.12×103Da,8.73×103Da and1.01×104Da(4) Infrared (IR) spectroscopy and nuclear magnetic resonance (NMR) spectroscopy were used determine the configuration of DR. The result showed that DR1was a kind of pyranose polysaccharides and a-configuration. DR2and DR3was β-configuration.(5) The result of antioxidant activity of polysaccharides from Dictyophora rubrovalvata showed that the crude polysaccharides with strong reducing power, at the same concentration conditions, the antioxidant activity of crude polysaccharides was investigated by using DPPH, hydroxyl radical and superoxide radical assay systems, the results showed that has scavenging effect. And with the increase of polysaccharide concentration, the scavenging more obvious.(6) The results of bacteriostatic activities of crude polysaccharides showed:the crude extract had a strong inhibition on Staphylococcus aureus, Escherjchia coli, Bacillus subtilis, Apple penicillium and Brewer's yeast. And the bacteriostatic activity for the Bacterial higher than the Mildew.