Studies On The Purification And Structure Analysis Of The Fruit Body Flammulina Velutipes Polysaccharides (FVP) And Their Evaluation Of Acetylcholinesterase Inhibitory Activity

The Flammulina velutipes is large edible macrofurgi, it has many aliases, such as plain mushroom, enoki mushroom, shiitake, needle mushroom, asparagus. Flammulina velutipes polysaccharide is a polysaccharide conjugation which was extracted from the fruit body of Flammulina velutipes, and it has many functions with antioxidant, anti-tumour, immunity adjustment, puzzle for children and so on. This article was divided into three parts, The first part is the establishment of filter system on acetylcholinesterase inhibitory activity; The second part is the purification and acetylcholinesterase (AChE) inhibitory activity of the polysaccharide fractions; The last part is the structure analysis of the purification fractions.AChE inhibitory activity was estimated using spectrophotometric method of Ellman. The results indicated that the optimized selecting conditions were the concentration of PBS 100mmol/L, the pH of PbS 10, the final concentration of AchE 0.05u/mL, the final concentration of ATCI 0.10mmol/L, the final concentration of DTNB 0.0667mmol/L, the conentration of Huperzine-A 70μg/L, the body reaction temperature 37℃, the reaction time 30min, and the proper wavelength 412nm.The FVP40 and the FVP60 were purified by the DEAE sapharose fast flow anion exchange column and the sephacryl S series gel chromatography, Thus the FVP40C-1 and the FVP60C-1 were obtained. The HPLC testing confirms that they were both homogeneous polysaccharides. Full-wave UV spectra indicated that both of them contain no protein and nucleic acids.The structure of FVP40C-1 and the FVP60C-1 were analysized by the methods of acetylation and methylation. The meta-analysis of the results as follow:FVP40C-1 was composed of fucose, xylose, glucose, mannose, galactose, molar ratio was 0.62:1.00:2.55:0.93:2.90. It was inferred that the backbone were 1,6-linked Glu and 1,6-linked Gal. The structure of mannose and xylose were complex because of a large amount of polysaccharide residues. FVP60C-1 was composed of glucose, mannose and galactose. And molar ratio was 1.54:1.00:0.68. The backbone were also 1,6-linked Glu and 1,6-linked Gal. The galactose was also acted as branched chain. The structure of mannose was also complex and need further studies by NMR measurement and other physical and chemical analysis.