Study On Separation And Activity Of Adenosine Substance In Submerged Fermentation Of Cs-4

This paper studies the liquid submerged fermentation of North Chinese caterpiller fungus(CS-4),as well as optimizes the fermentation process of 20 L fermentor by comparing content of adenosine compounds in CS-4 mycelia, to solve the problem of resources shortage of wild Cordyceps sinensis. The researches also optimizes extraction process of method-ultrasonic extraction method combined with microwave by the Box-Behnken Design, and analyses adenosine compounds with qualitative and quantitative character by UV spectrophotometry, TLC scanning method(TLCS) and high performance liquid chromatography(HPLC) method. Meanwhile, the study has been done to purify adenosine compounds through ion-adsorption method, combined with HPLC,besides, it investigates parts of biological activities of adenosine substance. The results are as follows:1. The experiment draws growth curve of CS-4 fungus using turbidimetry method,then optimizes the fermentation process of 20 L fermentor, and gets the best process: ventilation quantity is 1:1.0~1:1.2v/v·min, rotation rate is 180 r / min, temperature is 24.5~26.5 ℃, inculum concentration is 13%, initial pH is 6.0~7.0.Thus, the quantity of adenosine substance in CS-4 mycelia is 60.89ug/g. Some of the biological activities of the adenosine substance were also investigated.2. The research optimizes method-ultrasonic extraction method combined with microwave to extract adenosine compounds,by single factor experiments and the Box-Behnken Design,and gets the best conditions : extraction solvent is distilled water,ultrasound time is 15 min, microwave min is 6min, microwave power is 440 W, Solid-liquid ratio is 300mL/g.The extraction rate of adenosine compounds is 61.24%.3. It analyses the adenosine substance in CS-4 mycelia, and measures the contents of cytidine, uridine, adenosine, uridine, cordycepin is 92.54ug/g, 305.79ug/g, 1.79mg/g,785.85ug/g, 695.64ug/g, with regression equation, using UV spectrophotometry, TLCS and HPLC method.4. It optimizes purifitation process of adenosine materials, using 732 resin, through single factor experiments and the Box-Behnken Design, and gets the best process: velocity is 2.0BV/h, the sample pH value is 4.0, the concentration of sample is 1.6mg / mL, the quantity of sample is 0.8BV. And the best elution process are : elution volume is 4BV,elution concentration of ammoniae is 0.15 mo L/L, velocity is 2.5BV/h. The elution rate of Cordycepin also can get 94.52%,with method of recrystallization by prep-HPLC.5. The research studies antioxidant activity of adenosine compounds in vitro. Theresults show that adenosine compounds have the ability of antioxidant. It also studies antibacterial property of adenosine compounds and the effects of Staphylococcus aureus,Bacillus subtilis bacillus, Escherichia coli and actinomycetes etc. The results show that adenosine compounds can inhibite these pathogenic bacterias obviously in food.