Separation And Purification Analysis Of Soluble Polysaccharide By Submerged Fermentation Technology Of Morchella

In this experimentation, Morchella mycelium was prepared by submerged fermentation technology. Mensurated the extracellular polysaccharide from Morchella Produced by Liquid Fermentation, The polysaccharides from Morchella were extracted by ultrasound technology and Ultra-filtering Technology successively and determined the optimal extraction conditions. Finally, study the purification and characterization of water soluble crude polysaccharide.At aspect of determination method of polysaccharide, the mensuration of the extracellular polysaccharide of Toadstool may be interfered by the carbohydrate (such as starch ,sucrose) brought from culture medium. In this article, the extracellular polysaccharide are mensurated by the traditional method : the method of phenol– vitriol and the improved ROBERTS method. Experiment show the method of ROBERTS measured the polysaccharide content only occupies the method of phenol– vitriol to measure the polysaccharide content 17.3%. Experiments proved that ROBERTS Determination on extracellular Polysaccharide of Toadstool is more accurately.Study the ultrasonic extraction of polysaccharides from Morchella hyphostroma in this experimentation.The ultrasound extraction of Morchella hyphostroma polysaccharides was studied using single factor and orthogonal method. The optimal extraction conditions were determined as follows: the time of 20 min, liquid- solid ratio of 1:10 ( g: mL), extraction frequency of 2, ultrasound power of 960 W. The yield of the crude Morchella polysaccharides extraction ratio was 1.14%. This technology could decrease the costs of energy and time for industrial production.Using ultra-filtering technology to condense and extract extracellular polysaccharide of morchella.Determine the optimum condition in the process of using ultra-filtering technology to condense and extract extracellular morchella polysaccharide. Selecting retention rate and membrane flux as the parameter, using three different criteria such as ultra-filtering diaphragm's size, ultra-filtering pressure, ultra- filtering temperature to determine the best ultra-filtering condition.Found the best ultra-filtering condition to condense and extract extracellular morchella polysaccharide is:Use ultra-filtering diaphragm which has particle retention rate of 10KDa, ultra-filtering pressure should be 0.100Mpa, while the ultra-filtering temperature should be 20 degree Celsius. Under this ideal condition, extracellular polysaccharide's retention rate could reach 95.72%.The polysaccharides from Morchella hyphostroma and liquid fermentation were isolated, purified, and characterized. The crude polysaccharides were deprotinized by sevage method repeatedly, isolated and purified by DEAE -52 cellulose, Sephadex-100 gel filtration chromatography. The purity and molecular weight were identified by HPGPC. Their specific rotations were also determined and the sugar compositions of polysaccharides were analyzed by IR . Three crude polysaccharide fractions (PW1 ,PW2 and PW3) were extracted from liquid fermentation of toadstool; Two crude polysaccharide fractions (PN1, PN2) were extracted from Morchella hyphostroma. The polysaccharide fractions PW1,PW2,PN1,PN2 were homogeneous by gel filtration chromatography ,with the molecular weight were 13062,44259,39537 and 8300 respectively.