Studies On Enzymatic Extraction And Purification Of Ophiocordyceps Polysacchride From Paecilomyces Hepiali And The Optimization Of Purification Technique

Paecilomyces Hepiali is a typical endoparasitic fungi in natural Ophiocordyceps sinensis.Since its mycelial fermentation product has various health benefits and bioactivities,Paecilomyces Hepiali Cs-4 has been widely applied to medicines and functional foods in recent years.Polysaccharide is a main bioactive intergredients in Paeilomyces Hepiali and has also attracted substantial attention for its pharmacological functions including anti-oxidation,anti-radiation,anti-aging,immunomodulation,antitumor,as well as hypoglycemic and hypocholesterolemic effect.In our study,the extraction yield of alcohol precipitation Ophiocordyceps polysaccharide by the Pectinase PL was obviously superior to the extraction yield in water followed by ethanol precipitation.The polysaccharide yield would increase with the increase of enzyme dosage and enzymatic hydrolysis time,and the proper extraction time is 2.5-3.0 h.25 enzyme reagents were selected to polysaccharide extraction research,and the extraction yield is increased of 2.88-57.44%than that in control groups.Results show that the higher polysaccharide yield was obtained by Econase HCP,Pectinase PL and Protease NT group,which has increased polysaccharide yield by 53.90%,57.44%and 46.26%,respectively.The polysaccharide content in Econase HCP,Pectinase PL and Protease NT group is also superior to control groups.The polysaccharides yield by compound enzyme(Econase/Pectinase/Protease)was nearly doubled that of single enzyme,but the polysaccharide amount were not significantly different between single enzyme and combinative enzyme extraction.The result indicated that the enzymatic extraction effect of combinative enzyme is significantly better than that of a single enzyme.The polysaccharide was further purified with ethanol precipitation,and the higher extraction yield was increased with the increase of concentration ratio(extract solutions volume:dry weight of mycelia).The ration of 1.5:1(mL:g)has increased polysaccharide yield by 33.60%than the ration of 6.0:1(mL:g).Within the range of 0.5-6 volume ratios of ethanol,the yield increased with the increase of volume ratios of ethanol,but the yield decreased with the increase of volume ratios solutions to more than 6.3%active carbon was used for decolorization of polysaccharides under the optimum conditions:temperature 50-60?(water bath),time 50-60 min,add 1.5%white clay 5 min before the end of decolorization.The recovery rate of Ophiocordyceps polysaccharide is 76.13%and the decolorization rate is 82.24%under this condition.The three step filtration was applied to centrifugal supernatant with qualitative filter paper,0.80 ?m and 0.45 ?m microfiltration membrane,and the filtering effect is remarkable.The technical route of Ophiocordyceps polysaccharide extraction and purification for industrial production was established in our study.The application of combinative enzyme can efficiently increase the utilization rate of polysaccharide,reduce production cost and improve market competitiveness.