Studies On The Liquid Cultivation Of Corduceps Militaris And The Extraction And Functional Studies Of Cordycepin From The Fungus

Cordyceps militaris(L)Link is a precious medicinal and edible fungus.It contains various active ingredients and has anti-bacterial,anti-oxidant,protecting liver,anti-tumor,enhancing immunity and other bilogical activity.Cordyceps militaris is an alternative of traditional valuable Chinese medicine Cordyceps sinensis.Cordycepin is one of the main biological active substances in Cordyceps militaris.Cordycepin has anti-tumor,anti-virus,regulating immunity,interferencing the synthesis of mRNA and DNA and other bilogical activity.Cordycepin is one of the focus in the recent studies.In the studies the liduid cultivation conditions of Cordyceps militaris,and the separation and purification of codycepin from the liquid cultivation medium of Cordyceps militaris were studied.The mechanism of cordycepin inhibiting the growth of BGC823 cancer cells was also preliminarily studied.The liquid cultivation of Cordyceps militaris is the most common way to acquiring cordycepin recently.The previous studies of other authors show that the majority of cordycepin is in the liduid medium during the liquid culture of Cordyceps militaris.In the studies it was determined that the suitable cultivation medium of the Cordyceps militaris strain C2 was 3.0%glucose,2.0%peptone,2.0%yeast extract,0.1%KH2PO4,0.01%MgSO4·7H2O,0.01%CaC12,and pH6.5.It was showed that the dry weight of mycelia reached the maximum after cultivated for 6 days and the content of codycepin in liquid medium reached the maximum after cultivated for 7 days.The liquid culture medium of Cordyceps militaris,was concentrated by the flash evaporator for the first time.After the carbohydrate and protein was wiped out the liquid culture medium was appropriately concentrated by rotary evaporator,the cordycepin monomer was got using the conjuction of the 732 cation exchange resin column and silica gel chromatography separation.The obtained cordycepin conformed to the features of high performance liquid chromatography and UV wavelength scan detection,and its purity is 98.08%.The content of cordycepin in the Cordyceps militaris liquid culture medium is about 0.0525mg/ml.The ratio of cordycepin in the liduid culture medium and in the mycelium is 40:1.The huaman Gastric cancer cell line BGC823,hepatocellar carcinoma 7721 and cervical cancer Hela cells were exposed to the obtained cordycepin for different concentration and treating time.And the activity of thses cells was assayed by MTT method.It was showed that cordycepin could significantly inhibit the growth of these cells,and its inhibitory effect had exceeded the commonly used anti-cancer agent cyclophospamide,and just weaker than the cisplatin on the overall level.These results revealed that cordycepin had the pontention to be a broad-spectrum ant-cancer agent.The inhibiting effect of cordycepin to human hepatoma 7721 cells is especially strong,and the inhibiton rate could reach to 60.7%.It was found that apoptosis and autophagy could be induced when the BGC823 cells exposed to the 20μg/ml cordycepin.And autophagy of BGC823 cells induced by cordycepin was discovered for the first time.These studies provided a new research direction for the ant-tumor cells growth functions of cordycepin.