Optimization Of Culture Conditions And Biological Activity Of Sarcodon Aspratus

Sarcodon aspratus is a delicious mushroom distributed in Japan, Korea and China. It is a tricholomataceae fungus belonging to basidiomycotina and mainly composed of proteins and polysaccharides as chemicals components. This study aims to optimize the culture conditions and biological activity of S. aspratus.(1) optimization of the culture conditions by response surface methodology:The best source of carbon and nitrogen sources were glucose and yeast extract, respectively. Plackett-Burman designs were used to evaluate the major factors which had an important role in mycelium production. And the yeast powder, KH2PO4and cultivate time were the more significant factors. The optimal combined conditions were further optimized by response surface methodology. The results showed that the mycelium production could reached up to12.71g/L when the followed conditions were used. yeast powder:15.15g/L; KH2PO4:1.27g/L; cultivate time:4.17d; glucose:20g/L; CaCl2:1.2g/L; temperature:28℃; rotation speed:180r/min; liquid volume:200mL; inoculum size:3%. The mycelium production was improved46%compared that of previous.(2) Isolation, and antitumor activity of S. aspratus polysaccharides:Two polysaccharide fractions (neutral polysaccharide PSAN and acidic polysaccharide PSAA) were extracted and isolated by hot water. DEAE-Sepharose F.F anion-exchange chromatography and Superdex200. High Performance Liquid Chromatography (HPLC) analysis showed that the average molecular weight of PSAN and PSAA was approximately5.6×104Da and3.83×105Da, respectively. Structural properties and compositions were examined by HPLC and FTIR. PSAN was composed of rhamnose, xylose and mannose, with the molar ratio of0.1:1:2.1, PSAA consisted of rhamnose, xylose and mannose, glucose and galactose, with the molar ratio of0.11:4.3:8.4:1.1:2.3. It was conclued that they belong to β-type heteropolysaccharides with pyran group according to FTIR. Screening antitumor activities of polysaccharides SA was tested on there kinds of cancer cells, Hela, HepG2and As49cells. Further study showed that PSAA presented higher(70%) antitumor activity against Hela cells in a dose-dependent manner, and exhibited significantly lower cytotoxicity to human normal liver cell line L-02.(3) The effect of alcohol extract of S. aspratus on tau phosphorylation of AD: After treatment with different concentrations of ESA, p-tau of the OA-N2a cells decreased obviously (p<0.05). At the same time, the expression of p-GSK-3β and O-GlcNAc increased. Combining with the observation of cell morphology, we concluded that ESA had a protective effect on OA-induced AD cells. Fed mice ESA30days by intragastric administration before Hippocampal injection of Aβ, the p-tau of AD mice decreased obviously (p<0.05) Compared with the control. At the same time, p-GSK-3p and O-GlcNAc of hippocampus rised. Taking into account the observation of animation, it is concluded that ESA could prevent and resistance the AP-induced AD to a certain degree. Fed Aβ-induced AD mice ESA by intragastric administration, the p-tau, p-GSK-3β and O-GlcNAc of hippocampus had no significant changing compared with the model group. ESA possibly had no effect on therapy of AD.