Study On Submerged Fermentation And Selenium-enrich Technologcial Conditions Of Antrodia Camphorata Mycelium

Antrodia camphorata is a peculiar medicinal fungus in Taiwan, which has anti-oxidant and anti-tumor effect, but its wild resources are very scarce. Main research contents and results are as follows:The optimum submerged fermentation culture medium for Antrodia camphorata biomass, triterpene and polysaccharides respectively were studied by Plackett-Burman design and Response surface methodology (RSM) experiments. The optimum fermentation conditions were optimized by single factor test and Response surface methodology (RSM) experiments. The results showed that:for Antrodia camphorata biomass, the optimum culture medium is glucose 6.76%,yeast extract 0.67%, KH2PO4 0.1%, MgSO4·7H2O 0.05%, VB1 10mg/100mL; the optimum conditions were:temperature 28.58℃, filled volume 140mL/250mL, fermentation time 8.43d, initial pH2, oscillator speed 150rpm, seeding volume 15%; in this condition the biomass reached 6.78g/L; For triterpene, the optimum culture medium is Maltose 4.57%, yeast extract 0.45%, KH2PO 40.08%, MgSO4·7H2O 0.05%, VB, 10mg/100mL; The optimum conditions were:temperature 24.94℃, filled volume 113.69mL/250mL, fermentation time 8.98d, initial pH6, oscillator speed 150rpm, seeding volume 12%, triterpene reached 82.33mg/L. For polysaccharides, the optimum culture medium is soluble starch 3.17%, wheat bran 0.79%, KH2PO40.1%, MgSO4·7H2O 0.05%, VB1 10mg/100mL. The optimum conditions were:temperature 26.8℃, filled volume 110.24mL/250mL, fermentation time 8.4d, initial pH2, oscillator speed 150rpm, seeding volume 12%; polysaccharide reached 183.46 mg/ L.The study of Antrodia camphorata Selenium-enrich showed that NaSeO3 concentration will affect the growth of Antrodia camphorata,triterpene content and polysaccharides content, the concentration of selenium 20μg/g is optimum. The Se-enrich fermentation culture medium was optimized by Plackett-Burman design and RSM experiments:glucose 2.43%, soluble starch 2.34%, wheat bran 0.8%, KH2PO40.1%, MgSO4-7H2O 0.05%, VB1 73.17mg/L; the fermentation conditions were optimized by single factor test:temperature of 28℃, initial pH 6, seeding volume 10%, filled volume 100mL/250mL, oscillator speed 150rpm, the fermentation time 9d, In this conditons the selenium accumulation rate reached 58.65%, the total selenium content reached 856.676μg/g, biomass was 3.4g/L.The paper has established a systematic method to determine adenosine in Antrodia camphorata with high performance liquid chromatography. It could be separated completely on Agilen C18 column and detected at 260nm, using methanol: ultra-pure water=17:83 as the mobile phase at flow rate 0.8mL/min, the column temperature was 30℃. The linear range of Adenosine in Antrodia camphorata was within 3.9062-125μg/mL, R2=0.9997(n=6). The average recovery rate was 95.35% (RSD=1.07%).This method, reliable, simple, reproducible. The concentration of adenosine in Antrodia camphorata is about 26.086 to 42.348mg/g by this method.