Study On Structure And Bioactivities Of Polysaccarides From Ramaria Botrytoides

Ramaria botrytoides belongs to Aphyllophorales, also known as the broom fungus, which contains many kinds of amino acids, mineral elements, vitamins and carbohydrates. Early researches about Ramaria botrytoides polysaccharide focus on the extraction and the ability to remove the free radicals. The paper is about extraction, purification, physical and chemical properties,structure, immune, anti-tumor and antioxidant activity of Ramaria botrytoides polysaccharide, and wished to helping for the development of Ramaria botrytoides.The optimum extraction by hot water of Ramaria botrytoides Polysaccharide is determined by orthogonal experiment. The optimum extraction is as follows: 90℃,the ratio of material to liquid is 1:30, 3 times, each time for 60 min. Under the condition, the yield of crude polysaccharide is 5.24%. After the extraction of Ramaria botrytoides Polysaccharides,then decolorization, alcohol precipitation and protein removal are carried out, and the yield of RBP was 4.89%. DEAE-52 anion exchange chromatography was used to get the RBP-I, RBP-II, RBP-III, and with accounts for 16.50%, 28.17% and 7.17% respectively. Combination with GPC and G-100 Sephadex column chromatography, RBP-I is homogeneous polysaccharide, the molecular weight Mw is 15857 Da, RBP-II was miscellaneous polysaccharide, RBP-III is homogeneous polysaccharide, and Mw is 2188924 Da. With researches about physical and chemical properties,It was found that RBP-I, RBP-II, RBP-III are like white cotton, white cotton, yellowish cotton respectively,all them do not contain starch and protein contents are 0, 0, 1.03%, total sugar contents followed by 86.94%, 91.45% and 81.07%,uronic acid contents followed by 5.14%, 7.77%, 4.87%.Structure analysis: RBP- Ⅰ consists of rhamnose, fucose, arabinose, mannose, glucose, galactose and molar ratio is 1: 6.53: 1.99 8.29: 70.05: 5.38; RBP-Ⅲ is made of mannose, glucose and galactose, and molar ratio of corresponding monosaccharide is 4.76: 4.61: 1. The infrared spectra show that RBP-Ⅰand RBP-Ⅲ are polysaccharide containing β-D-pyran ring and mannose glucose. By periodate oxidation and Smith degradation, RBP-Ⅰcontaining1→6, 1→2,1→4 and 1→3 glycoside bonds, the molar ratio is 112.23:1:5.46, and linked with 1→3 glycosidic linkage mainly are glucose; RBP-Ⅲ contains 1→6, 1→2,1→4 and 1→3 glycoside bonds, the molar ratio is 6.18: 1: 2.58, and linked with 1→3 glycosidic linkage mainly are mannose, glucose and galactose. By NMR analysis, RBP-Ⅰis made of six kinds of monosaccharide and containsα- pyranose andβ-pyranose two configurations, the main chain is 1→6 glucosidic linkage; RBP-Ⅲ is made of three kinds of monosaccharide, for α-pyranose andβ-pyranose configurations, containing1→6, 1→2,1→4 and 1→3 glycoside bonds.The biological activity of Ramaria botrytoides polysaccharides: the polysaccharide samples have antioxidant activity, and are related with the concentration of polysaccharide. The DPPH and ABTS+ free radical scavenging capacity of RBP-Ⅰis better than that of other polysaccharide samples, crude polysaccharide,s ability of reducing Fe3+ is the strongest; ORAC measured antioxidant capacity RBP-Ⅲ> RBP-Ⅱ>RBP> RBP-Ⅰ. The inhibitory effect of Ramaria botrytoides polysaccharides on HepG-2 and MCF-7 cells is positively correlated with the concentration. RBP-Ⅲis best, comparable to the positive control 5-Fu uracil, RBP-Ⅰand RBP-Ⅱis better than RBP about antitumor activity. The Ramaria botrytoides polysaccharide alone in mouse spleen lymphocytes can promote cell proliferation, but the degree of the improvement is lower, when LPS and ConA synergistic effect on cells, RBP-Ⅰ, RBP-Ⅱ, RBP-Ⅲ and RBP can promote proliferation, but four has no significant difference. Ramaria botrytoides polysaccharide can promote the secretion of NO,IL-6 and TNF-α, and has a dose dependent effect. RBP-Ⅰis the strongest, followed by RBP-Ⅲ, RBP-Ⅱ and RBP. RBP-Ⅰand RBP-Ⅱ, RBP-Ⅲ and RBP can promote the expression of IL-6 and TNF-αgenge of RAW264.7, improving cellular immune activity.