| In this paper, we successfully get the fruiting bodies of Cordyceps militaris. Then we use the fruiting bodies as material, establish a method of analysis the content of adenosine and cordycepin in Cordyceps militaris by high performance liquid chromatography. On the basis, this paper change the carbon, nitrogen, vitamin types, pH value of nutrient solution, culture medium and the amount of pupa powder, to explore the relationship of culture factors and fresh weight, dry weight, polysaccharides, proteins, adenosine, cordycepin content in the bodies of Cordyceps militaris. The main conclusions are as follows:(1) In the experiment of passaged strains, F0-F5 Dynasties strains were inoculated in solid medium culture, although the first three generations appeared bacteria degradation in the process of culture, but three generations later, the ability of the strains to generate fruiting bodies of Cordyceps species have recovered. Observe the sporulation of subcultured Cordyceps sinensis,we found generations hyphae have serial production spore structure, sporulation was not observed.(2) A HPLC method for determining the contents of adenosine and cordycepin in Cordyceps militaris(L.) is described, and its extraction conditions were optimized comprehensively. The separation column was XBridgeTM C18(5 um,4.6 × 250 mm), which was held at 30℃. The mobile phase was composed of pure water and methanol(V/V,84:16) at a flow rate of 1.0 mL/min. The sample loading volume was 10 μL. UV detection was performed at 254 nm. The blank-spike recoveries of adenosine was 99.13% and those of cordycepin was 99.18%. The add-spike recoveries of adenosine in 3 replicates determinations was 95.78% and those of cordycepin was 94.35%. The limit of detection was 0.01 μg/mL for both adenosine and cordycepin. The two components was extracted by pure water, under 30 ℃ and 700 W ultrasonic power for one time. This method is characterized by simplicity and high sensitivity and accuracy.(3) Nitrogen source types, pupa powder and culture medium have a more significant impact on the fresh weight and dry weight of Cordyceps militaris. Comprehensive fresh weight and dry weight, the peptone is the best nitrogen source. When the nitrogen source is wheat bran, the militaris yield(fresh weight 16.32 g, dry weight 3.80 g)is lower than peptone, but it is also significantly higher(fresh weight 13.78 g dry weight 3.02 g) than the control group, and its low price will have more economic benefits; 0.5 g pupa powder per bottle cultivation medium can get 7.06 g fresh weight and 1.37 g dry weight than the control group, but with the increase of the content of pupa powder, The fresh weight and dry weight of Cordyceps militaris did not increase; When the culture are millet(27.16 g, 5.01 g) and wheat(25.26 g, 4.20 g), Cordyceps militaris can have a higher fresh and dry weight.The factors of Nitrogen source, vitamins and culture media can obviously improve the polysaccharide content in Cordyceps militaris. When the nitrogen source is peptone, vitamin is VB1, culture medium is rice, it is more conducive to the improvement of polysaccharide content in Cordyceps militaris. When nitrogen source is soybean meal, culture medium is sorghum rice or corn, the protein content in Cordyceps militaris will be higher.Whe the carbon is source Glucose, the adenosine content in Cordyceps militaris is the highest(169.5 mg/100 g), followed by maltose(163.1 mg/100 g). Whe the glucose is the carbon source, the content of cordycepin(149.0 mg/100 g) is the highest, considering the two indicators of adenosine and cordycepin, glucose is the best carbon source; When the bran and soybean meal as nitrogen source, the adenosine content in Cordyceps militaris is(184.6 mg/100 g),(178.8 mg/100 g) and the cordycepin content is(172.8 mg/100 g)(146.9 mg/100 g). Therefore, the best nitrogen source is wheat bran, the followed one is soybean meal; When the corn is culture medium, the content of adenosine(248.5 mg/100 g) is highest, followed by wheat(225.9 mg/100 g), and when rice is medium, the content of adenosine(186.1 mg/100 g) is the lowest. When the sorghum is medium, the cordycepin content(248.2 mg/100 g) is the highest; When the pupa powder is 0.5 g, the adenosine content(204.4 mg/100 g) is higher than that of the control group(179.6 mg/100 g),but it did not increase with the improvement of the pupa powder. |
PDF Full Text Request