Study On The Response Of Different Extraction Methods To The Structure And Activity Of Chroogomphus Rutilus Polysaccharides

Chroogomphis rutilus is a precious edible medicinal fungus under the forest.Polysaccharide is an important active ingredient in Chroogomphis rutilus,which has anti-tumor,anti-oxidation,anti-fatigue,blood sugar reduction multiple pharmacological effects.Extraction is a key link in the research and utilization of natural polysaccharide resources.However,there are few reports on its extraction methods and structure-activity relationship.Therefore,this paper optimizes the four extraction methods for the extraction of polysaccharides from Chroogomphis rutilus,and analyzes the antioxidant and immunoregulatory activities of the four polysaccharides.At the same time,the structural characterization of the four polysaccharides is studied.The research provides the theoretical basis,and the results are as follows:(1)The single-factor combined response surface test was used to optimize the extraction process of polysaccharides from Chroogomphis rutilus by traditional water extraction,ultrasonic extraction,microwave extraction,and ultrasonic microwave extraction.Among them,the ultrasonic and microwave extraction method has a high extraction rate.It has the advantages of short extraction time and low solvent consumption.It is the best method for extracting polysaccharides from Chroogomphis rutilus.The optimal process conditions are:liquid-to-material ratio 29:1 mL/g,ultrasonic time 8 min,microwave time 6 min,and microwave temperature.92℃.Under these conditions,the extraction rate was 8.69%±0.19%.(2)The results of physicochemical analysis of polysaccharides obtained from the four processes show that the polysaccharide of Chroogomphis rutilus is compsed of D-pyranose glucose,mannose and galactose,mainly composed of→6)-α-D-Glcp-(1→and→4)-α-D-Glcp-(1→triple helix heteropolysaccharide,different extraction methods did not change the glycosidic bonds,main functional groups,and monosaccharide types of the polysaccharide of Chroogomphis rutilus,but could significantly affect its monosaccharide composition,molecular weight,triple helix conformation and appearance;there is no significant difference in the chemical composition of the four CRPs,and there is a certain amount of sulfate groups.In terms of molecular weight,the highest molecular weight of CRP-UM is 145 kDa,and the lowest molecular weight of CRP-U is 121 kDa.In addition,all four CRPs have a triple helix conformation,and CRP-UM has a stronger forming ability.(3)The in vitro antioxidant and cellular antioxidant activities of the polysaccharides obtained by the four extraction methods were investigated.The results showed that the four polysaccharides of Chroogomphis rutilus had certain antioxidant activities,of which CRP-UM had the strongest total reducing power,·OH,ABTS+and ·O2-radicals scavenging ability,and has the strongest alleviating effect on H2O2-induced HL-7702 fine oxidative damage,can significantly improve cell viability,increase its T-SOD and GSH-Px activity,reduce Cell MDA content.(4)The results of comparative analysis of in vitro immunomodulatory activities showed that the polysaccharides obtained from the four extraction methods can promote the activation of RAW264.7 macrophages in a dose-dependent manner,promote the release of NO,and enhance its cytokines TNF-α and IL-6 secretion,increase ROS levels in cells.Unlike the antioxidant results,CRP-H has the strongest activation ability among the four polysaccharides,followed by CRP-M and CRP-U,and CRP-UM has the weakest activation ability.