The Separation, Purification, Modification And Properties Of Alkaline Dissolved Polysaccharide From Bean Dregs

Bean dregs polysaccharide has anti-aging, antioxidant, aperient, reducing blood fat,preventing colon and rectal cancer effects on the human body, and also has many functionalproperties, including the viscosity, emulsification, foamability and oxidation crosslinking andso on.With bean dregs as raw materials,this paper systematically studiedextraction,isolation,purification,modified of the bean dregs polysaccharide,and the physicaland chemical properties,functional properties and antioxidant activity were discussed.Themain research contents and conclusions are as follows:(1) Through testing the influence of different ultrafiltration parameters on theconcentration of concentrated polysaccharide solution and membrane flux,and ultimately theoptimum conditions of ultrafiltration technological for the bean dregs polysaccharideextracted by the alkaline H2O2method as follows: pressure0.08MPa,temperature45℃,pH6.5.On this condition, the concentration of concentrated polysaccharide solution is25.94mg/mL, membrane flux is53.32mL/min.(2) Deproteinization of bean dregs crude polysaccharide was studied.Through comparingthe deproteinization effect of enzymatic method,Sevage method, the enzymatic combinedwith Sevage method,TCA method, enzymatic combined with TCA method, the test resultsshowed that the enzymatic combined with TCA method is the best and polysaccharide lossrate is relatively lower.The optimum conditions of deproteinization for enzymatic methodwere as follows:the enzyme solution temperature50℃, enzymolysis time2h, pH8.0, addingenzyme quantity1.5%. Adding TCA into enzymatic hydrolysis of crude polysaccharidesolution to make the final concentration be1.5%, in this condition,protein removal rate isabout90%, polysaccharide loss rate is about40%.(3) After extraction bean dregs crude polysaccharide was purified on DEAE SepharoseFast Flow column, and tested the effects of the different elution mode,different ionconcentration, different elution velocity, different sample quantity and the concentration ofthe sample on the separation and purification of bean dregs polysaccharide.The experimentalresults showed that,the sample quantity of8ml,sample concentration of10mg/ml,elutionvelocity of2ml/min, and in this condition, with distilled water,0.1M sodium chloride,0.3Msodium chloride eluted the column separately.Three components were washed down and respectively named AHPSPSW,AHPSPS0.1, AHPSPS0.3.Because AHPSPSW activity is nothigh,only AHPSPS0.1and AHPSPS0.3components were studied. And on this condition, thebean dregs polysaccharide extrated by alkali method also were purified and got threecomponents, respectively named ASPSW,ASPS0.1and ASPS0.3(4) AHPSPS0.1,AHPSPS0.3,ASPS0.1and ASPS0.3four components were furtherpurified with Sephacryl S-300by column chromatography experiments, and tested the effectsof the flow rate,sample concentration and the amount of sample on purification ofpolysaccharide.The experimental results showed that the flow rate is0.5ml/min, samplequantity is2ml, sample concentration is6mg/ml.On this condition,the eluted solution ismainly a single component.(5) Sulfuric acid ester bean dregs polysaccharide were prepared by chlorosulfonicacid-pyridine, the best reaction condition obtained by orthogonal test is: chlorosulfonic acidand pyridine volume ratio of1:3,esterification reagent and polysaccharide solution volumeratio of4:3,reaction temperature is80℃,the reaction time is2.5h.On this condition,the degreeof substitution of bean dregs polysaccharide is up to2.15. And the sulfating polysaccharidewere purified and obtained two components, respectively named S-AHPSPS0.1and S-AHPSPS0.3(6) This paper preliminary studied the rheological properties, oxidation crosslinking,emulsion stability and antioxidant activity of the bean dregs polysaccharide(AHPSPS0.1,AHPSPS0.3)extracted by alkaline hydrogen peroxide, the bean dregs polysaccharide(ASPS0.1, ASPS0.3)extracted by alkali method and sulfuric acid esterification bean dregspolysaccharide (S-AHPSPS0.1,S-AHPSPS0.3). The experimental results showed that they allhad a certain viscosity properties, and the viscosity increased with the increase of bean dregspolysaccharide solution concentration and sucrose adding quantity, decreased with theincrease of temperature andshear rate, while the amount of pH and salt concentration had littleimpact on the viscosity of the solution; Concentration, The different added amount of pH andsalt ions had a certain degree of influence on emulsification of various polysaccharidessolution, but slightly; Various polysaccharide solution all had a certain removal effect onsuperoxide anion free radicals and DDPH·, but it was not very strong, and the hydroxyl freeradical scavenging effect was the best;The ability of50%removing hydroxyl free radical（EC50）for AHPSPS0.1,AHPSPS0.3,S-AHPSPS0.1, S-AHPSPS0.3、ASPS0.1and ASPS0.3was9.45mg/ml,8.20mg/ml、6.62mg/m,6.33mg/ml,15.25mg,mland10.56mg/ml，respectively.