The Effect Of Ethanol Extract Of Gastrodia Elata On The Components And Hypoglycemic Activity Of Extracellular Polysaccharides Of Grifola Frondosa

This arcticle aims at studying the effect of the ethanol extract of Rhizoma gastrodiae(R.gastrodiae)on the composition and hypoglycemic activity of exopolysaccharides(EPS)from Grifola frondsa(G.frondsa)by submerged culture.Firstly,to achieve the purpose of preliminarily purification of exopolysaccharides from G.frondsa,the method of protein removal process was optimized.Secondly,different fermentation medium was added to the fementation system and then using anion exchange column chromatography separation of components of EPS from G.frondsa.In this way,we can study on EPS component changes with the main active ingredients of R.gastrodiae involved in G.frondsa fermentation medium.Finally,the main ingredients of R.gastrodiae were added in G.frondsa fermentation medium.And analysed its EPS effect on ?-glycosidase enzyme activity inhibition rate.Thus,we can infer that the effect of the main ingredients of R.gastrodiae on the EPS hypoglycemic activity from the perspectives of enzymology.Experiment of the main research conclusions as follows:Protein removal rate and polysaccharide loss rate were used to be the index of evaluating and comparing the effectiveness of Savage method,trichloroacetic acid(TCA)method,papain method in deproteinizing EPS from G.frondsa.The result showed that TCA method was the most effective method with the best addition amount 3%.In this condition,the Protein removal rate was 68.40% and the polysaccharide loss rate was 21.11%.What's more,few absorption peaks were observed by the UV-visible scanning spectrum between 200-600 nm,this indicated that TCA method would be one of the best useful method to remove protein which included in the crude EPS from G.frondsa.The main effective ingredients of R.gastrodiae including R.gastrodiae extracts,p-hydroxybenzyl alcohol(HA)and p-hydroxylbenzaldehyde(HBA)were involved in G.frondsa submerged fermentation to obtain four EPS called GFP-1?GFP-2?GFP-3?GFP-4 respectively.These four EPS were eluted natural polysaccharose EPS-1 by ultrapure water.With 0.2 mol/L NaCl,it was eluted the acidic polysaccharose calledEPS-2 and eluting the EPS-3 acidic polysaccharose by 0.5 mol/L NaCl.However,GFP-2,GFP-3,GFP-4 were eluted EPS-4 acidic polysaccharose by 0.5 mol/L NaCl,and EPS-4 in GFP-4 increased at most.Thus,we can infer that the effective ingredients of R.gastrodiae can make the amount of EPS of G.frondsa redistribution between components,and in this process,HBA had the most effective on the components of EPS in the metabolism of G.frondsa.The main effective ingredients of R.gastrodiae were involved in G.frondsa submerged fermentation to obtain GFP-1?GFP-2?GFP-3?GFP-4.Analysis GFP-1?GFP-2?GFP-3?GFP-4 and acarbose on ?-glycosidase enzymes were half inhibitory concentration(IC50)from big to small order: GFP-1 > acarbose > GFP-4 > GFP-3 > GFP-2.Hypoglycemic activity from high to low order as: GFP-2 > GFP-3 >GFP-4> acarbose > GFP-1.This indicated hypoglycemic activity of EPS got by conventional fermentation culture of G.frondsa was lower than the clinical drug acarbose.But when the effective ingredients of R.gastrodiae including R.gastrodiae extracts,HA and HBA were involved in G.frondsa submerged fermentation respectively,hypoglycemic activity of EPS increased higher than that of clinical drug acarbose.Among these,the R.gastrodiae extracts was the largest contribution component of the synergistic action.