Studies Of The Russula Fermentation Optimization And Some Biological Activity Of Polysaccharides

Objective:Determine the fermentation conditions for the Russula mycelium fermentation; detect inhibit ability Russula polysaccharide to Staphylococcus aureus, E. coli,and mildew; disclose polysaccharide of Russula adjustment to senile rat regulation of immuno organs; testing different doses of Russula polysaccharides induced TLR mRNA on macrophage.Methods and Results:1.Optimization experiments of Russula deep liquid fermentationBy phenol sulphuric acid method measured mycelium extract polysaccha-ride2.56 mg/ml, optimum carbon source was sucrose,3.0% sucrose fermen-tation is best,when the biomass reached exopolysaccharide the best value, > 3.0% or <3.0%, both down. Best nitrogen source is peptone, when the pep-tone concentration was 0.2%, the fermentation of the best, continue to impro-ve the nitrogen concentration, the fermentation decreased. When the MgSO4, KH2PO4 were 0.2% and 0.1%, fermentation is best, biomass was 9.21 mg/ml exopolysaccharide was 2.58 mg/ml. Orthogonal analysis of the influence of Russula fermented, sucrose> MgSO4> KH2PO4> peptone, sucrose is the main factor.2. Antibacterial activity of Russula mycelium and mediumThe antibacterial activity of Russula polysaccharides on G bacteria, such as E. coli better, than the G+ bacteria, which may have a dense peptidogly-can structure. Compared antibacterial effect of Russula polysaccharide, when PH was 2,4,6,8, found in PH 6 the inhibitory effect was best. Confirmed in different temperatures antibacterial activity of Russula polysaccharide, resul-ts above 60℃or more, decreased inhibitory effect. In MIC study,20 g/L of polysaccharides inhibited the E. coli proliferation,40g/L can inhibit many pathogens, to G-bacteria better than the G+. Russula mycelia has clear antibacterial, particularly strong inhibitory activity against E. coli. Zone of inhibition of the 24,72 hours of observation, the antibacterial effect was continuing. Inhibition of Russula polysaccharides on bacterial growth curve, inhibit the proliferation of the logarithmic phase with significantly. The Russula polysaccharide may inhibit key division of bacterial. Russula polysaccharides also inhibit fungal spore germination.3.Russula polysaccharide immune balance homeostasis testThymus, spleen index increased has dose-dependent polysaccharide. In 20 mg/g polysaccharide group the thymus index was 3.75, spleen index was 6.02 and close the young group level. In antibody formation experiment Russula polysaccharide 20mg/g significantly increased the antibody levels in senile rats, close to the level of young rats. Polysaccharides inter-vene spleen lymphocyte apoptosis percentage from 24.34% to 12.47%, with the control group senile rats significantly reduced. Suggesting that Russula polysaccharides can significantly interfere with excessive apoptosis of lymphocytes.4. The activation mechanism of polysaccharides on macrophageRussula polysaccharide significantly increased peritonealmacrophage phagocytic index, in particular 100 ug/ml group was 38.37%, phagocytic index of 0.67, close to the LPS-induced rate. In different polysaccharide concentrations, the experimental group were there TLR4 gene expression. TLR antibody significantly reduced the LPS group, IL-1 expression, where as a weak expression of polysaccharide group. NF-κB Changes with TLR and IL-1 has synchronous relationship, presumably polysaccharide induced in macrophages TLR-IL-NF-κB chain reaction.Conclusion:1 In mycelial of Russula submerged fermentation process the best carbon source was sucrose, nitrogen source was peptone,and sucrose concentration most importantfermented. Obtained Russula polysaccharide by submerged fermentation feasible.2 Russula mycelial and fermented liquor has the antibacterial activity on pathogenic bacteria and fungi, Russula polysaccharides as biological agents can assist the clinical treatment of pathogeninfection. 3 Polysaccharide can be corrected to the reduction of immune cells, fading of synthesis functional. Russula polysaccharide promote mother cellsof lympho-cyte, regulate T cell synthesis DNA, the active cellular immune potential. Polysaccharides can increase the humoral immune response tomaintain im-mune homeostasis.4 Polysaccharide activated TLR receptor on macrophages,started TLR-IL-1-NF-κB path, activated macrophages the synthesis reaction, maybe it's a one of the mediation role of the path.