Study On Isolation, Purification And Biological Activity Of Polysaccharides From Clitocybe Maxima Quél

Fungi polysaccharides were natural macromolecule compound derived from the sporlcarp or mycalium of the fungus.They had such a lot creatives as antiviral property, anticoagulant property,putting down blood-lipid,against cancer,immunomodulation and slowing down the process of getting older,so they have been developoed as many kinds of medicines and additives for foods.With the further stude on natural drug chemistry and medical theory,the research of fungi polysaccharides has caused great interests of scholors from home and abroad,their important biological effects have become the hotpoint.Polysaccharides from Clitocybe maxima Quél(ZDG) was precious fungi for food,it has the significant value in science and application.In this study,we explored extraction,isolation and purification conditions on polysaccharides from Clitocybe maxima Quél.Furthermore,we investigated the biology activity of the polysaccharides with the mice,which include immunomodulation,anti-fatigue and anti-oxidation.As a result,the research about medicine base on the polysaccharides from Clitocybe maxima Quél is very important.The results were as follows:1) Sample pretreatmentPretreatment of the Clitocybe maxima Quél(ZDG) polysaccharide's sample is important.The sample added to 70%ethanal solution and stayed 30min in ultrasonic instrument,then deposited 24h.Pretreatmenth prosessing is very useful not only inhance content of ZDG polysaccharide but also wipe off protein,it was beneficial to isolate and purify ZDG because of reducing impurity.2) Extracting ZDG polysaccharide with hot waterThe optimization process conditions of extracting temperature,ratio of solid-liquid, the duration of time and the extracting times in extracting polysaccharides from ZDG,has been undertaken by orthogonal experiment of L₉(3⁴).And optimum technological conditions were as follows:temperature was 90℃,the extracting time was 4h,the solid-to-liquid ratio was 1:30 and the extracting times was 3.3) Decoloring with active carbonDosage,temperature and time of decoloring were investigated during processing of decoloring with active carbon,we found that the suitable content of active carbon was 1%,and optimal extracting time was 60 minute.4) Purification of ZDG polysaccharide with Macroporous ResinAfter choosing from nine macroporous resin,we selected AB-8 to purify ZDG polysaccharide.The optimal eluting solution was 0.2mg/mL NaCl,and the best flow of elution was 2mL/min.5) Isolating Clitocybe maxima Quél polysaccharide with DEAE-celluloseZDG was separated by DEAE-cellulose chromatography.The different eluting solution was used to washing the column,then got three frations-one neutral polysaccharide(ZDG-1) and two acid polysaccharides(ZDG-2 and ZDG-3).6) The monosaccharides composition of ZDG-2Single sugar composition of ZDG-2 was studied in details by GC methods,the results showed it consisted of D-seminose,D-galactose and glucose.7) Biological activity study of ZDG polysaccharide:Biological activity of ZDG was studied in mice,which included immune regulation, anti-oxygen and anti-fatigue.The results proved that ZDG is effective in enhancing immune,anti-oxygen and anti-fatigue.