Fermentation Process Optimization,Oral Liquid Preparation And Biological Activities Of Cordyceps Militaris

Cordyceps militaris and cordyceps sinensis are from the same genus, but belongto different kinds of typical, also known as north cordyceps sinensis. Reports showthat cordyceps militaris can be used as the new fungus medicine source instead ofthe cordyceps sinensis.There are several active ingredients of cordyceps militaris,one of them is called cordyceps polysaccharide, many studies suggest that it hasmany effects, such as anti-aging、enhancing immune system and lowering bloodsugar. The development of health food oral liquid is an important direction, andespecially for the development of fungi polysaccharide oral liquid. This thesismainly studies from four aspects, the optimization of cordyceps militarisfermentation conditions, the preparation of oral liquid, stability and toxicity analysisof oral liquid, antioxidant of oral liquid and immune-modulating activity.Optimization research for cordyceps militaris strains in the experiment of shakeflask culture conditions. First, from the Plackett-Burman design experimentsanalysis, we get seven conditional factors of speed, age, temperature, cultivationtime,inoculation amount, fluid volume, initial pH, and initial pH、 culturetemperature、time had a great impact on expectations. Further studied from the Box-Behnken design experiment and a multiple quadratic regression analysis tooptimize the optimal values of the three significant factors, finally determine theoptimum of the cordyceps militaris strains shake flask culture conditions for: initialpH of5.4, temperature of25.7℃, incubation timeof5.9d,3d of age, the speed of150r/min, the quantity of5%, fluid volume100ml/250ml conical flask. Thecordyceps militaris strains fermened under this condition, with the largestexpectations of0.5175, which is increased by13.08%compared with the notoptimized culture conditions, and the dry weight of mycelium, adenosine,polysaccharide and cordyceps acid production of the bacteria was increased by4.33%,11.76%,15.17%and6.41%respectively.Adjusting the concentration of the polysaccharide solution of cordyceps militaris for5mg/ml extracted from the optimal conditions. In order to increase thestability of cordyceps militaris polysaccharide oral liquid, first added a certainamount of stabilizer and preservatives in the process of oral liquid preparation of hintin this paper, according to the experimental data, after filtration, to get cordycepsmilitaris polysaccharide oral liquid, sterilization by10ml of brown bottle filling;Second, we made experiment of the light effect, temperature effect test andaccelerated test to the cordyceps militaris polysaccharide oral liquid in this paper。Then the toxicity of the oral liquid test was carried on, acute toxicity experiment inmice at the same time we also do the subacute toxicity experiments in rats.Above all, the experimental results showed that the stability of cordycepsmilitaris polysaccharide oral liquid is good,with minor side effects.Finally, we conducted a series of experiment on oral liquid in vitro, in vivoantioxidant: oral liquid scavenging effect on the OH; Different concentrationgradient oral liquid on O2-scavenging effect; Different concentration gradientoral liquid on the inhibition of oxidative hemolysis of red blood cells induced byH2O2; Different concentration gradient of the oral liquid on the inhibition of lipidperoxidation of liver homogenate in mice;The subacute aging model was establishedbased on kunming mice, using the mice liver, brain tissue and serum, textingdifferent oral liquid of SOD activity and MDA content and gsh-px activityrespectively.Hands-free regulating experiment on low immune function model inmice is built up by using cyclophosphamide. To prove its immunomodulatoryactivity by comparing the mice NK cell activity by the blank group, model groupand experiment group. And use the same group to test the mice’s content ofserum hemolysin and the immune organ index, to further improve its activeimmunity experiment.The experimental results show that, oral liquid prepared in thispaper has a certain of oxidation resistance, also immunity activity at the same time.In this paper we get the best culture conditions for cordyceps militarissuccessfully. The polysaccharide oral liquid has a good stability, security, and theactivity of pharmacodynamics. It provided a reliable preliminary experimental basis for the development of cordyceps militaris oral liquid industy and the increase of theproduction.