Preliminary Study On New Technology Of Antrodia Cinnamomea Active Product Fermentation Coupled Extraction

Antrodia cinnamomea is a rare medicinal fungu which is unique in Taiwan.It only parasitizes on the inner wall of the heartwood of fallen dead Antrodia cinnamomea,which is difficult to obtain.Antrodia cinnamomea has a strong medicinal effect,the unique ubiquinone compound 4-acetylantroquinonol B(hereinafter abbreviated to 4AAQB)in its mycelium has a significant inhibitory effect on hepatoma cells Heph2 and can be used as a potential anticancer drug.However,due to its growth conditions,large-scale production cannot be achieved.In this experiment,mycelium was obtained by liquid fermentation culture to improve the shortcomings of the long production cycle and unstable quality which are caused by the cultivation of fruiting bodies.In the liquid fermentation,a new fermentation coupled extraction process is used,so that the original intracellular products flow out of the cell and are enriched by the extractant in the fermentation broth to achieve preliminary separation.The main work with the following content:1.By investigating the types,add time,and amount of various external additives such as benzene ring-containing substances,the addition of 0.008%(m/v)p-toluic acid on the sixth day of the fermentation process was screened out successfully,As a small molecular substance,adding low concentration solution can significantly increase the 4AAQB and triterpenoid compounds in Antrodia camphorata mycelia,the yields are 0.57 mg/g and 130.26 mg/g,which are 3.35 times and 1.61 times higher than the blank control group.It was first obtained in the current research that it can avoid the factors of the large amount of exogenous additives and complicated separation which could cause subsequent separation difficult.It has an important role in the research on the metabolic pathway of 4AAQB and triterpenoids synthesized by Antrodia camphor ata.2.The experiment conducted the preliminary researches of the surfactant and extractant types,addition amounts and add time in the 4AAQB system by fermentation coupled extraction.The optimal conditions obtained from the experiment were that 2%(v/v)Tween-80 was added on the 0th day of the fermentation process,and 30%(v/v)oleic acid was added on the 16th day.The intracellular 4AAQB yield was 0.011 mg/g,the organic phase yield was up to 1.6 mg/g,and the total yield was 26.67 times that of the blank group,significantly increasing the yield of 4AAQB in the unit system.3.The experiment conduct the preliminary researches of the surfactant and extractant types,addition amounts and add time in the triterpenoid system by fermentation coupled extraction.According to the research results,the optimal condition is to add 2%(v/v)Tween-80 on the 0th day of the fermentation process and 30%-50%(v/v)oleic acid on the 16th day.The yield of intracellular triterpenoids is 64.2 mg/g,and the yield is 12g/L in the organic phase,which greatly improves the yield of triterpenoids in the unit system.The optimal conditions are similar to the above process,it can be considered to use uniform conditions to produce these two products at the same time.4.Combined with the above optimal conditions,a study on using both p-toluic acid and fermentation coupled extraction system which means that add 2%Tween-80 on the 0th day of fermentation,add 0.008%p-toluic acid on the 6th day and add 30%oleic acid on the 16th day of fermentation can obtain 0.09 mg/g of 4AAQB and 210 mg/g of triterpenes in the cell,6.84 mg/g of 4AAQB and 6.94 g/L triterpenoids in the organic phase.Compared to using these two methods alone,the target product 4AAQB have a significant improvement.