The Effect Of Gastrodia Elata And Fagopyrum Tataricum Compound Solution On The Synthase And Antioxidant Activity Of Grifola Frondosa Extracellular Polysaccharide

In this paper,the effect of adding Gastrodia elata and Fagopyrum tataricum compound solution to the fermentation system of Grifola frondosa on the extracellular polysaccharide synthesis enzymes of Grifola frondosa and the antioxidant activity of polysaccharides.The researches mainly included that adding Gastrodia elata and Fagopyrum tataricum alcohol extract in different mass concentration to the submerged fermentation of Grifola frondosa to determine the optimal ratio of the compound solution,and dynamically determine the changes of key enzyme activities during fermentation;Using the DPPH radical scavenging rate,hydroxyl radical scavenging rate and ABTS radical scavenging rate as indicators,the effects of additives on the antioxidant activity of Grifola frondosa extracellular polysaccharides were investigated;The mice were fed with Grifola frondosa fermented broth cotain Gastrodia elata and Fagopyrum tataricum compound solution,and the antifatigue effect of the fermentation broth was investigated using the exhaustive time of weight-loaded swimming,the hepatic glycogen level,blood ureanitrogen,etc.as indicators,The metabolomics method of UPLC-QTOF-MS was used to study the metabolic differences of adding compound solution on Grifola frondosa fermentation.The main findings are as follows:Using the extracellular polysaccharide production and bacterial biomass of Grifola frondosa as indicators,the results showed that the best effect was observed after additon of Gastrodin elata and Fagopyrum tataricum at concentrtions of 7g/L and 5g/L resperctively,the extracellular polysaccharide production and bacterial biomass increasing 49.08% and 51.10% compared to the blank group,both were significantly higher than the blank group(P <0.01)and the single addition group(P <0.05).The addition of Gastrodia elata and Fagopyrum tataricum compound solution can enhance the activity of phosphoglucomutase and UDP-glucose pyrophosphorylase,while reducing the activity of phosphoglucose isomerase,which was increased by 86.98%,69.12% and decreased by 65.78% compared with the blank group.Both phosphoglucosemutase and UDP-glucose pyrophosphorylase showed maximum activity at the 7th day of fermentation(P <0.05),and glucoglucose isomerase showed the lowest enzyme activity at 7d(P <0.05)The experiment results of antioxidant activities in vitro showed that Grifola frondosa polysaccharide has good antioxidant activity,and within a certain mass concentration range,the free radical scavenging rate is positively related to the polysaccharide concentration,and it shows a significant concentration dependence,but are lower than VC..Among them,the polysaccharides of Grifola frondosa added with Gastrodia elata and Fagopyrum tataricum compound solution had the best effects on DPPH radical scavenging rate,hydroxyl radical scavenging rate and ABTS radical scavenging rate,which were increased by 11.05%,14.28% and 11.96% compared with the blank group.Experimental study on anti-fatigue in mice showed that gavaged the mice Grifola frondosa fermentation broth can improve the exhaustive time of weight-loaded swimming,the hepatic glycogen(HG)level,and lactate dehydrogenase(LDH),reduce the contents of lactic acid(LA)and urea nitrogen(BUN),and the higher the concentration of the fermentation broth,the more obvious the effect.At the same dose of gavage,the fermentation broth with Gastrodia elata and Fagopyrum tataricum compound solution was significantly higher in anti-fatigue performance than the blank group(P<0.05)and the single-addition group(P <0.05).The metabolomics method of UPLC-QTOF-MS was used to study the metabolic differences of Gastrodia elata and Fagopyrum tataricum compound solution fermented Grifola frondosa.The results showed that the principal component(PCA)model showed significant differences in metabolites between the two groups(P<0.05).Through orthogonal partial least squares discriminant analysis(OPLS-DA),44 differential metabolites were screened and identified under the conditions of VIP>1 and P<0.05,including 6 surgars,13 amino acids,5 vitamins,7 nucleotides,10 organic acids and 3 fatty acids.Analysis of metabolic pathways of different metabolites by Metabo Analyst 4.0 to obtain 14 key metabolic pathways.Further analysis showed that amino acid and nucleotide sugar metabolism,pentose and glucuronic acid conversion,galactose metabolism,TCA cycle,vitamin metabolism,alanine,aspartic acid and glutamate metabolism and other metabolic processes were accelerated in Grifola frondosa mycelium cells after adding the compound solution.Experiments showed that the addition of Gastrodia elata and Fagopyrum tataricum compound solution promotes the extracellular polysaccharide synthesis of Grifola frondosa,changes the key enzyme activity,enhances the polysaccharide antioxidant activity,and improves the nutritional quality of the fermentation broth.