| Gomphidius Rutilus is a kind of precious edible fungus. The research on Gomphidius Rutilus mainly focused on its cultivation technique and mycelium fermentation and the study of polysaccharides is still not deep enough. Along with the rapid advancement of the community, fatigue is particularly common in the life. So, how to alleviate and eliminate fatigue has gained great popularity and it is of great significance. The polysaccharides of fungal has attracted much attention because of it’s important physiological biological activity. This paper, taking Gomphidius Rutilus as raw material, extracted, purified and identified the polysaccharides and carried out a detailed study on its alleviating physical fatigue. This study has laid a theoretical basis for future in-depth research and utilization of Gomphidius Rutilus polysaccharides, and provided a reference for food development.(1) The extraetion of polysaccharides from Gomphidius Rutilus and the optimization of proeessThe polysaccharides of Gomphidius Rutilus(GRP) were extracted by Water solution and alcohol sedimentation. The optimum conditions of extraction was investigated through response surface methodology. The results were as follows: ratio of water to raw material,20:1; extraction temperature, 90℃; extraction time, 2.5h; and extraction frequency,3. Under these conditions, the experimental yield of crude polysaccharides was 7.73%.Protein in the Polysaccharides was removed by Sevage method. After application of this method to remove 6 times, protein removal rate can reach 97.71%.(2) Purification and classification of polysaccharides from Gomphidius RutilusDEAE-cellulous chromatography and Sepharose CL-6B gel column chromatog- raphy was used for the isolation and purification of the crude polysaccharides. After isolated and purified by DEAE-cellulous chromatography. Four components GRPA-0, GRPA-1,GRPA-2 and GRPA-3 were got. Total sugars and uronic acid assay, only GRPA-0 does not contain uronic acid. GRPA-1、GRPA-2 and GRPA-3 isolated and purified by Sepharose CL-6B gel column chromatography, Four components GRPA-2-1, GRPA-2-2, GRPA-3-1and GRPA-3-2 has been gained. The four components all content uronic acid.(3) The analysis of structures of four polysaccharides fractionThe four components were studied through UV wavelength scanning. The peak of 190 nm was agree with the absorption peak of characteristics of polysaccharides. No obvious absorption peak at 260 nm and 280 nm, which shows that there was no free nucleic acid and protein in the polysaccharides. Infrared spectrum scanning results showed that the four components contains pyranose ring. Monosaccharide composition analysis by highperformance liquid chromatography, The results showed that GRPA-2-1 and GRPA-2-2were composed of mannose, rhamnose, galacturonic acid, glucose, galactose and fucose.The ratio of Man, Rha, Gal UA, Glc, Gal and Fuc was 4.4:2.3:1.8:53.3:28.1:10.1 and5.3:1.9:3.1:51.1:30.2:2.3, respectively. GRPA-3-1 and GRPA-3-2 were composed of mannose, rhamnose, galacturonic acid, glucose, galactose and arabinose. The ratio of Man,Rha, Gal UA, Glc, Gal and Ara was 4.3:8.2:1.2:72.1:8.6:5.6 and 3.4:3.5:2.1:78.2:9.9:2.9, respectively. The average molecular weight of four components were 440550 Da,687610Da, 377650 D and 1118550 Da, respectively.(4) The study of alleviate physical fatigue of GRPThe anti-fatigue experiment was studied in mice. Feeding the mice with GRP of low(100mg/kg/d), middle(250mg/kg/d) and high(625mg/kg/d) dosage, Determinate the exhaustion swimming time, the content of blood lactic acid and blood urea nitrogen and hepatic glycogen.It showed that three dose groups of GRP can extend the swimming time of mice, improve the mouse athleticism. The effect of middle dose is remarkable.compared with control group, biochemical index determination results shows that the GRP dose group can significantly reduce the content of blood urea nitrogen and blood lactic acid,increase the content of hepatic glycogen. To some extent, the GRP has the function of relieving physical fatigue. |
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