The Artificial Cultivation Of Cordyceps And The Study Of Extraction And Purification With Cordycepin

In this thesis, the north Cordyceps fungus (Cordyceps Militarism) was chosen as the object of research, and the optimization of culture conditions of its liquid fermentation were studied systematically. The yield of cordycepin mycelium was as evaluation indexes. In the optimized culture mediums, the fungus of the north Cordyceps were enlarged to cultivate under the optimized culture conditions, then analyze its growth rhythm. Optimize the process of extraction and purification of Cordycepin in the fermentation broth, and make the obtained crystal of cordycepin by qualitative and quantitative analysis. The results are as follows.1. Through the growth curve of the north Cordyceps fungus, get the best inoculation of seed age for72h.2.Study Cordyceps fermentation medium carbon, nitrogen species and suitable concentration, and levels of orthogonal experiment for the best carbon source, the best nitrogen source, inorganic salts and vitamins for inspection analysis, the best Cordyceps fermentation bacteria based formula as follows:glucose20g/L, peptone8g/L, KH2PO4: MgSO4=1:1, dosage1g/L, VB11g/L.3. Analysis of inoculum (%), initial pH, fermentation temperature (℃), stirring rate (h) through the single-factor test. Analysis on above the four factors as the basic.Through the orthogonal experiment. Determine the optimum fermentation conditions were:inoculum is10%, initial pH=7,the temperature of fermentation is27℃, stirring rate is150r/min.4. Through nine factors and two level climbing experiment of comprehensive development of conditions of north cordyceps sinensis bacteria, the result shows in the99%confidence area. Then we can know that the important factor of cordycepin grain weight just as follows:the quantity, the initial pH, glucose addition amount and add volume of peptone.5. Based on the four main factors gotten in the climbing experiment, the response surface center combination regression model is set up.Then we can obtain good fitting degree mathematical model by the data integration,including model P<0.0001.The results showed that the model can be used to predict the north Cordyceps sinensis bacteriain content in different culture conditions.6.In the optimize training conditions in north from cordyceps sinensis to increase training, extracting cordycepin in Chinese caterpillar fungus fermented liquid. Cordyceps content is as an indicator to determine the optimum extraction method is ultrasonic alcohol formulation. 7. Orthogonality optimize the process of Ultrasonic extraction of Chinese caterpillar fungus, ensure extraction solvent is:water:ethanol (V/V)=1:3, extracting time30min, extraction times for3times, extraction power for800W, getting the highest content element to Chinese caterpillar fungus1.447g/L.8. Based on the study of purification of cordycepin to determine the purification column for the cation resin, the column flow rate of solution2.0ml/min, a volume of800ml; best washing dosage1000ml; best to wash ammonia concentration0.2mol/L,ammonia volume of800ml, flow control2.0ml/min.9. Respectively using UV, IR, high-performance liquid chromatography, mass spectrometry and nuclear magnetic resonance methods and standards of cordycepin cordycepin purified crystal structure identification, test results showed that the purified crystals identified is cordycepin.