Studies On Artificial Culture And The Total Triterpenes Producing In Antrodia Camphorata

Antrodia camphorata is a parasitic fungus that is found only in Taiwan under the name “niu-chang-ku”.A number of bioactive components have been identifided from Antrodia camphorata.Triterpenoid has been recognized as one of the most potential bioactive components.The problems that hangting researchers are the methods of cultivating Antrodia camphorata fruitbody and the low production of triterpenoid of Antrodia camphorata in liquid fermentation.In this study,the cultural characteristics of Antrodia camphorata on solid media and liquid media were studied.The culture condition and culture media of Antrodia camphorata was optimized to increase the triterpenoid production.Antrodia camphorata fruiting body was cultivated with different aging log.The total triterpenes producing in Antrodia camphorata was analized and effects of extracts from different Chinese herbal medicine on growth and intracellular triterpenoids formation of Antrodia camphorata was studied.To explore the cultural characteristics of Antrodia camphorata on solid media,Antrodia camphorata was cultured on solid agar medium.Effects of different media,carbon sources,and nitrogen sources on growth and morphology of Antrodia camphorata were observed,and the effects to the content of total triterpenes of Antrodia camphorata were explored.The growth velocity and morphology of Antrodia camphorata were different in various media.Carrot media,PDA media and Wheat bran media were suitable for solid culture in Antrodia camphorata.The content of total triterpenes of Antrodia camphorata on carrot media and GPY media are higher than in other media.Different carbon sources could be utilized by Antrodia camphorata.Antrodia camphorata grew better when carbon sources was fructose,maltose,mannose,xylose,xylitol in submerged culture conditions.The content of total triterpenes of Antrodia camphorata was higher when carbon sources was glucose,maltose,mannose.Better adaptability to nitrogen source except for urea were observed in Antrodia camphorata.Antrodia camphorata was not grew when nitrogen source was urea.The growth velocity and morphology of Antrodia camphorata were different in various nitrogen source.The content of total triterpenes of Antrodia camphorata was higher when nitrogen sources was ammonium sulphate and fish peptone.To explore the cultural characteristics of Antrodia camphorata on liquid media,Antrodia camphorata was cultured on different liquid medium.Effects of different media on growth and the content of total triterpenes of Antrodia camphorata were observed.In order to increase the triterpenoid production,the culture condition of Antrodia camphorata was optimized by method of one-variable-at-a-time.The optimized result was showed as following: cultural time 20 d,the pH 4,media volumes 150 mL,speed 120 rpm.The carbon sources,nitrogen sources and growth factors of Antrodia camphorata was optimized also by the method of one-variable-at-a-time.The optimized medium was carbon sources Xylose,nitrogen sources NaNO3 and growth factors Corn steep liquor.Studies on artificial culture of Antrodia camphorata fruiting body,the old apple wood,pear wood,chestnut wood,and oak wood were cut into a certain length of pile.These woods were immersed in water for 7 to 10 days and inoculated with Antrodia camphorata.Vegetative growth and reproductive growth management were needed.Eventually fruiting body grew on the abandoned old apple wood.Then pilot scale production for Antrodia camphorata mycelia in submerged culture was experimented in a 50 L stirred-tank bioreactor.Working volume of the 50 L stirred-tank bioreactor was 30 L.Seeding culture was carried out at 28? for 3 or 5 days.And inoculate the Antrodia camphorata to the apple wood.Vegetative growth and reproductive growth management were needed.Eventually fruiting body grew on the abandoned old apple wood.The aim of the research is to investigate the quality and the content of the total triterpenes in different perioid in Antrodia camphorata mycelia.Antrodia camphorata was cultured on different solid agar medium.The content and the changes regulation of the total triterpenes in Antrodia camphorata mycelia are measured.High-performance liquid chromatography is employed to analize the kinds of the total triterpenes in Antrodia camphorata mycelia.And the study researched on the changes regulation of the total triterpenes in different medium and period.With an addition of decreasing in carrot media from 50 to 60 day,the content of the total triterpenes rises from starting to the end on different solid agar medium.The kind and the content of the total triterpenes were demonstrated certain regularity and variance in different periods of the same media or in different medium of the same period.The aim of the research is to investigate the effects of water and ethanol extracts from different Chinese herbal medicine on growth and intracellular triterpenoids formation of Antrodia camphorata.The Antrodia camphorata was inoculated into the basal media with and without addition of water and ethanol extracts fro m different Chinese herbal medicine,and then cultured for 20 d.The results showed that all the water extract had stimulatory effects on the mycelial growth except the water extract from Semen coicis in the concentration of 0.6%.And the water extract from Fructus lycii had significantly stimulatory effects on both the mycelial growth and intracellular triterpenoids formation of Antrodia camphorata.The ethanol extracts from Pericarpium citri reticulatae and Radix angelicae sinensis showed the inhibitory effects on the mycelial growth and intracellular triterpenoids production of Antrodia camphorata.However,ethanol extracts from Fructus forsythiae showed positive effects on the production of triterpenoids.