Structure Characterization And Lipid Activity Studies Of Polysaccharide Isolated From Sugarcane Residue

BackgroundPolysaccharide is composed of a dozen or even hundreds of monosaccharide molecules bound together by glycosidic bonds.Macromolecular compound carbohydrates and their derivatives are called polysaccharides.Polysaccharide is widely found in animal cell membranes and in the cell walls of plants,micro-organisms,and is one of the four basic materials of life,closely related with the maintenance of vital functions.By source category,it can be divided into fungal polysaccharide and bacteria polysaccharide,algae polysaccharide,plant and animal polysaccharide.Polysaccharides are important biologically active macromolecules,a wide study shows that polysaccharides have significant biological activity such as immune,anti-tumor,reduce blood sugar,lipid,radiation resistance,anti-bacterial antiviral,anti-oxidation,protecting liver and liver function.Research in polysaccharides has drawn world-wide attention in recent years.Sugarcane is an annual or perennial C4 plant of high photosynthetic efficiency native to tropical and subtropical areas.Generally,extract polysaccharides from sugarcane leaves and sugarcane residue.Early study indicates that polysaccharide of sugarcane residue has hypoglycemic effects.In recent years,research has shown that polysaccharides from sugarcane residue can significantly improve immune function in immunosuppressive mice.Polysaccharide of sugarcane leaves has a strong active to clear · OH and anti-tumor activity in vitro.Sugarcane residue is a by-product of the sugar industry belongs to one of agricultural solid waste,most of which has not been effective application.Modern pharmacological studies had showed that Sugarcane residue polysaccharides had immunomodulation,anti-tumor,antioxidant,hypoglycemic activities,and so on.Therefore,the studies on chemical structure of polysaccharide of sugarcane residue,activity and structure-activity relationship will be of great significance.Objective1.Study on the extraction process of degreasing,removal of protein from polysaccharides of sugarcane residue.2.Extraction and refinement of polysaccharides from sugarcane residue3.Isolation and purification of polysaccharides from sugarcane residue.4.Study on molecular weight and structure identification of polysaccharide obtained from sugarcane residue.5.Study on sugarcane residue polysaccharides for lipid activity of rats with hyperlipidemia.Methods1.Screening processOptimize of degreasing process by using uniform design.Contrast Sevage method and enzymatic effect of removal protein.2.Extraction of polysaccharides from sugarcane residueSugarcane residue,ethanol and ethyl ether(1:9)prepare,heat,to degrease,get rid of the liquid,add water twice,first time is about 10 times of the amount of filter residue,the second is 8.Then do the extraction twice.Condensing filtrate,add 4 times of 95%ethanol,filter and dry it.Join one-fifth volume of Sevage solution(chloroform:n-butyl alcohol 5:1)to crude polysaccharide solution shake 10min,take away the supernatant,then add one-fifth volume of Sevage solution,repeat 8 times.Add 30%H2O2 to polysaccharide solution,incubate 24 hours under 37 degrees.Put the polysaccharide solution in dialysis bags,dialysis 72h in distilled water,change water every 12h.Refined crude polysaccharides were extracted after freeze drying.3.The isolation and purification of polysaccharides from sugarcane residueSeparation of sugarcane residue polysaccharides solution by DEAE-52 column elution,collect eluent by automatic part of collectors.Use phenol-sulfuric acid method to track polysaccharide peak,and collect polysaccharides derived from separating.Use Sephadex G100 to further purify,water is used as eluent.Then purify samples on Sephacryl S300 to get higher purity,using Sephacryl S300 to verify the purity.4.Study on molecular weight and structure of polysaccharides from sugarcane residueUse Sephacyl S-300 gel column chromatography to determine the molecular weight.Use high performance ion chromatography to determine the monosaccharide composition after a full acid hydrolysis of sugarcane residue polysaccharides samples.Treat samples by partial acid hydrolysis,periodate oxidation-Smith degradation,methylation analysis and a series of chemical treatments.Combine with gas chromatography,GC detector,infrared spectroscopy,nuclear magnetic resonance to analyze the structure of polysaccharides.5.Study on the lipid-lowering effects of crude polysaccharides of sugarcane residueEstablish rat model of hyperlipidemia by feeding a high fat diet.After dose 4 weeks in a row,collect their abdominal aorta blood,testing TC,TG,HDL-C,LDL-C level.Results1.Studies of sugarcane residue degreasing process have shown that,the best degreasing process is:ethanol and ethyl ether:10%ethanol and cent ether;solid to liquid ratio is:1:25(g/mL),extraction time is 4h.By comparing the Sevage method of removal of protein and enzyme removing protein,shown that Sevage method of removal protein is more suitable.2.We get SRP1,SRP2 two polysaccharides after separation of sugarcane residue polysaccharides with DEAE cellulose column 52.With Sephadex G100,Sephacryl S300 for further purification,the result is a single symmetric peak.Validation with Sephacryl S300,the peaks are single symmetric peaks,shown that polysaccharides groups are pure.Detection of SRP1,SRP2 polysaccharide content by phenol-sulfuric acid are:93.2%,94.9%.3.SRP1,SRP2 polysaccharide molecules are:19,340;75,232.SRP1 is composed of Arabinose,Galactose,Mannose,the mole ratio is 9:5:15.SRP2 is composed of Galactose,Glucose,Mannose,the molar ratio is 8:10:9.4.Experiments have shown that polysaccharides of sugarcane residue can lower serum total cholesterol(TC),triglyceride(TG)and low density lipoprotein(LDL-C)concentration,increase high density lipoprotein(HDL-C)concentration.It has shown that the polysaccharides of sugarcane residue have lipid-lowering effect.Conclusion1.In the process of extracting polysaccharide,degreasing and removal protein processing can increase the rate of extraction.The water extracting and alcohol precipitation method for extracting polysaccharides has many advantages,such as:low energy consumption,simple operation,and suitable for industrial production.2.DEAE-52 cellulose,Sephadex G100,Sephacryl S300 can achieve satisfied effect on the isolating and purifying of polysaccharides from sugarcane residue.In addition,we obtained the polysaccharides structure units by a series of analyses.3.The activity of the rat as an animal model studies has shown that sugarcane residue polysaccharides have a good lipid-lowering effect.