Extraction,Purification And Lowing Plasma Cholesterol In Vitro Of Polysaccharide From Armillaria Mellea

In this study,Armillaria mellea polysaccharide was extracted with ultrasonic-assisted and further separation and purification was used.The reagents and conditions was explored.Armillaria mellea powder,after purified armillaria mellea polysaccharide Z1,Z2 and lentinan were studied by analysis of its hypolipidemic in vitro,according to this method,it provide a solid theoretical basis for developing of Armillaria mellea polysaccharide in the future.(1)In order to study the extraction technology of polysaccharide from Armillaria mellea,based on the polysaccharide yield,polysaccharide was extracted by ultrasonic-assisted enzymatic(cellulase,papain)method.This paper studied the optimal extraction conditions by single factor experiment,such as enzymolysis temperature,ultrasonic power,ultrasonic time,liquid material ratio,enzymolysis time,ratio of compound enzyme,enzyme concentration.The optimal conditions were obtained by response surface methodology on the basis of single factor experiment.By single factor experiment,enzymolysis temperature 50 ?,ultrasonic power 360 W,ultrasonic time 20 min were determined.The results of response surface methodology showed that the optimal extraction conditions were enzyme concentration 1.75%,ratio of compound enzyme 2:1,enzymolysis time 140 min,liquid material ratio 30:1(mL/g).Under the optimal extraction conditions,the yield of polysaccharide reached 40.56%.(2)Adding Sevage reagent to Armillaria mellea polysaccharide solution which should be removed protein,based on the deproteinization and polysaccharide yield.The optimal conditions were chloroform-butanol ratio 5:1,sample-Sevage ratio 5:1,deproteinization times 3,time 30 min.(3)Adding hydrogen peroxide reagent to Armillaria mellea polysaccharide solution which should be decolorated,based on the decoloration and polysaccharide yield.The optimal conditions were temperature 50 ?,pH8.0,time3 h,hydrogen peroxide dosage 40%.(4)The solution of Armillaria mellea polysaccharide was separated by ethanol fractional precipitation and further purified by gel chromatographic column.Then Armillaria mellea polysaccharide Z1,Z2 would be obtained.Purified polysaccharide were used in ultraviolet spectrum,show that it does not contain protein,amino acids and nucleic acid in the final product.Using infrared spectrum analysis of its structure,Armillaria mellea polysaccharide Z1,Z2 are?-pyran polysaccharide;By high performance liquid chromatography(HPLC)method to analyze monosaccharide composition of Armillaria mellea polysaccharide Z1,Z2.Z1 is composed of galactose,mannose,rhamnose,glucose,arabinose,fucose;Z2 is composed of galactose,mannose,rhamnose,glucose,fucose.(5)By hypolipidemic experiments in vitro,the blinding capacity of sodium cholate of Armillaria mellea powder,after purification of Armillaria mellea polysaccharide Z1,Z2.According to the same dosage and corresponding to simvastatin which is the main component of hypolipidemic and lentinan,the results showed that the blinding capacity of after purification of Armillaria mellea polysaccharide Z1,Z2 were superior than lentinan by different treatment.To sum up,confirm Armillaria mellea polysaccharide has good ability to hypolipidemic.