Effects Of Endophytic Fungus AJ14 On The Active Components Of Acanthopanax Senticosus

Fermentation technology has many years history in our country. Taking advantage of fermented traditional Chinese medicine by microbial can increase the active ingredient contents and the efficacy of TCM, expand the scope of the medicinal, reduce side effects, protect the environment and improve the taste. It has great significance to protect endangered species of wild resources.Acanthopanax senticosus(Rupr. et Maxim.)Harms, a frequently used herb, which is on the edge of extinction. In this work, Acanthopanax senticosus was fermented by endophytic fungi in order to study the contents changes of active components in the fermented products and the non-fermented Acanthopanacis senticosus.In this study, Acanthopanacis senticosus was chosen as substrate which was fermented by single strain and mixed strains with Acanthopanax senticosus endophytic fungi AJ7, AJ14, AJ15 and AJ19, respectively. And the dynamic changes of main active ingredients in non-fermented Acanthopanax senticosus and fermented samples were observed.Conclusions were as follows:1. The contents of isofraxidin were used as an index, and Acanthopanacis senticosus was fermented by single strain and mixed strains with Acanthopanax senticosus endophytic fungi AJ7, AJ14, AJ15 and AJ19. The results showed that the contents of isofraxidin in Acanthopanacis senticosus fermented by AJ14 were obviously higher than the others samples, so the Acanthopanacis senticosus endophytic fungi AJ14 was selected as target stran.2. Acanthopanacis senticosus was fermented by endophytic fungi AJ14, and three kinds of active ingredients (isofraxidin, syringin and polysaccharide) were chosen as indexs, then the dynamic changes of three kinds of active ingredients in non-fermented Acanthopanax senticosus and fermented samples were observed in order to analysis the effects of fermentation on Acanthopanacis senticosus.3. The process of fermented Acanthopanax senticosus by AJ14 was optimized by single-factor test combined with response surface method with the the contents of isofraxidin in fermented Acanthopanax senticosus as marker. The optimal process conditions were as follows:the substrate concentration was 0.2 g mL-1,the added volunm of AJ14 suspension(1×107 FU·mL-1) was 6mL, pH value was 7.5, fermentation temperature was 27℃, speed of shaker was 140 r·min-1, fermentation volumn was set at 30%, the fermentation time was 19d. Under the optimal conditions, the contents of isofraxidin was 0.6669 mg·g-1 in fermented sample, compared with the crude drug samples increased 8.07 times.4. The isofraxidin in non-fermented Acanthopanacis senticosus and fermented product were extracted, separated and purified. Isofraxidin was extracted by using heat reflux with 95% ethanol, and it was purified by TLC, silica gel column chromatography, then it was adnalyze by HPLC. In this way,115.45 mg isofraxidin were got in fermented Acanthopanacis senticosus and its purity was 93%.5. Endophytic fungi AJ14 strain was identified by morphological characteristics and molecular biology. The results indicated that endophytic fungi AJ14 strain belongs to Aspergillus ochraceus.