Optimization Of Liquid Fermentation Process Of Hirsutella Sinensis And Its Polysaccharide Preparation

Cordyceps sinensis （Cordyceps （Berk） Sacco.） is the the ergot Division（Clavicipiraceae） Cordyceps genus （Cordyceps）, mainly produced in Qinghai, Tibet,Yunnan, Guizhou, Sichuan ect, which altitude in3500⁵000m mountain meadow soil,herbs is the complex of Cordyceps fungus child seat and its host Cordyceps bat moth（Hepialusarmoricanus Oberthur） larvae corpse, rich in physiologically active substances,it is the valuable Chinese herbal medicines for the Chinese specialty. The paper usedstrain is Hirsutella sinensis. And the Hirsutella sinensis proved to be validly publishedCordyceps anamorph.Polysaccharide macromolecular active compound is polymerizationed by aplurality of monosaccharide or a derivative thereof, is the essential component of allliving organism, and is closely related to the maintenance of biological function.Polysaccharide is nonspecific immunomodulatory agents, having the biological functionof anti-tumor, anti-inflammatory, anti-clotting, anti-viral, anti-radiation, lowering bloodsugar, lowering blood presure ect, polysaccharide is becoming one direction of today’sdrug development[1-2].Based on the research object of Hirsutella sinensis and the target of dry weight ofmycelium, we optimize the Hirsutella sinensis fermentation conditions and medium;based on the goal of extracted amount of extracellular polysaccharide of Hirsutellasinensis, we optimize the preparation of Hirsutella sinensis polysaccharide. CultivatingHirsutella sinensis by composite medium in a10L fermentor, and seperatingfermentation mycelia and fermentation liquid, and then dried the mycelium, Theseparation liquid is first concentrated,and second extracting exopolysaccharides, andthird going through the progress of pre-column derivatization HPLC analysis.In order to increase the yield of Hirsutella sinensis mycelium, the first step isto optimize the fermentation conditions.Using the synthetic medium that PDA plusfifteen thousandths silkworm chrysalis meal to shake flask fermentation the Hirsutellasinensis, the explored optimum fermentation conditions: medium initial pH6.5, fluidvolume80mL/250mL, temperature18°C, shaking speed150r/min. The secondstep,optimized the Medium composition by single factor and orthogonalexperiments,and explored the final fermentation medium: glucose5g/L, corn flour15g/L, peptone15g/L,silkworm chrysalis meal5g/L,milk80mL/L,NH4Cl5g/L, KH₂PO₄0.8g/L, MgSO₄·7H2O0.3g/L, riboflavin （VB₂）0.8mg/L. Using thismedium culturing9d in10L fermentation jar，getting the dry weight of mycelium14.1g/L, the dry weight of exopolysaccharide16.47g/L. Both the Hirsutella myceliumand polysaccharide production have been greatly improved.Extracting Hirsutella sinensis polysaccharide with95%ethanol, drawning that4times the volume of ethanol extract the highest amount. Making the crudepolysaccharide been bleached, then deproteinized, and then going through the progressof precolumn derivatization HPLC analysis, derived the Hirsutella sinensis extracellularpolysaccharide monosaccharide composition is glucose, mannose, galactose.