Study On Liquid Fermentation And Fruiting Body Cultivation Of Cordyceps Militaris

Cordyceps militaris (L.) Fr, is a fungus used as tonic medicine and food,has the very high medicinal and nutritional values. Cordyceps militaris is the siblingspecies of Cordyceps sinensis, as well as the genus pattern of Cordyceps, which isused as Cordyceps sinensis in northern China folk. As the substitute of Cordycepssinensis, Cordyceps militaris has aroused people’s high attention. Modern researchhas shown that cordycepin is an important active ingredient of Cordyceps sinensiswith the function of anti-tumor, anti-virus, anti-fatigue, anti-microbial, antioxidant,improving the body’s immune. Furthermore, the contents of Cordyceps militaris’active ingredients, such as cordycepin, cordyceps polysaccharide, superoxidedismutase (SOD), adenosine, are significantly higher than the natural Cordycepssinensis. Hence, Cordyceps militaris is regarded as the ideal substitute of Cordycepssinensis.This study includes, firstly, establishing the standard curve equation. Thequantitative of cordycepin in the culture medium without fruiting body was measuredunder the optimized cordycepin ultrasonic extraction condition and HPLCquantitative analysis condition in order to provide a basis for reasonable utilization ofwaste solid medium. Secondly, different concentrations of vitamin B1, B6and2,4-Dwere added to Cordyceps militaris liquid fermentation medium and the effects ofmaterials on the production of cordycepin were studied. Thirdly, conduct theexperiment of Cordyceps militaris fruiting body’s cultivation. The effect of solidmedium composition,2,4-D, humectant, harvest time on fruiting body wereinvestigated.The main results are as follows:1. Established the cordycepin’s high performance liquid chromatography (HPLC)rapid detection method, the method can not only make the peak of cordycepin wellseparate with other peaks, but also can fast analysis on the cordycepin’s qualitativeand quantitative. This method also has a great value in Cordyceps militaris liquid fermentation and solid cultivation regarding cordycepin as indicator. Thedetermination of cordycepin in solid medium without fruiting body showed that, inthe5batches of Cordyceps militaris solid medium (can’t producing fruiting body),the average content of cordycepin reach561.32μg/g.2. The results showed that the addition of vitamin B1, B6and2,4-D obviouslyimproved the production of cordycepin. Moreover, both optimal levels for vitamin B1and B6were0.83g/L, and the best level of2,4-D was0.015mg/mL. The effects ofhigh temperature sterilization of vitamin B1, B6and2,4-D on the cordycepinproduction were studied. The results showed that the high temperature sterilizationdecomposed vitamin B1partly. Addition of vitamin B1before high temperaturesterilization was more effective to the increase the cordycepin production than aftersterilization and thus cordycepin was increased19.9%. It seemed that the decomposedproducts have better effect than the original vitamin B1.3. In this experiment, the conditions of Cordyceps militaris solid cultivation wereoptimized. The selection of solid material showed that, from inoculation to finish theturn color, the medium with rice and wheat1:1had the highest content of cordycepin,up to318.6μg/g. Because of the positive correlation on the content of cordycepinbetween the solid medium and fruiting body, the rice/wheat1:1was choosed as themedium of Cordyceps militaris solid cultivation. The experiment of addition of2,4-Dshowed that, adding2,4-D (6mg/L) is most suitable for Cordyceps militaris fruitingbody’s growth, and obtaining the maximum output5.8±0.3g/bottle, but too much2,4-D can lead to the abnormal growth of fruiting body. Adding humectant (nutrientsolution/humectant300mL:1g) can effectively reduce the times of water, and thefruiting body growth better. In case of the harvest time, during40-55days, the growthof fruiting body is in a good state, but after55days, the growth is nearly terminal andthe increasing of production is not obvious, thus, the best harvest time is55days.Additionally, this paper summarized the technologies of the solid cultivation ofCordyceps militaris and the cases needing attention.