Studies On The Active Metabolites Of CJ7 In Endophytic Fungi Of Acanthopanax Senticosus

Acanthopanax senticosus (Rupr. et Maxim)Harms, a northern native drug, which has many kinds of bioactive constituents, has been identified as an endangered medicinal plant by the government because of excessive exploitation. Many studies confirmed that endophytic fungi in plants could produce the same or similar active metabolites as bioactive constituents of their hosts. As a result, developing and utilizing the endophytic fungi resources in medicinal plants will bring significant ecological and economic benefit. In this paper, active metabolites of Acanthopanax senticosus endophytic fungi CJ7 will be studied in order to obtain the same or similar active metabolites as bioactive constituents of Acanthopanax senticosus and find a new method to preserve Acanthopanax senticosus resource and develop new medicine.CJ7 strain was identified by morphological characteristics and 18SrDNA sequence analysis. The results indicated that CJ7 strain belonged to Aspergillus Fumigatus.Meanwhile, fermentation conditions were optimized by mono factor experiment and orthogonal experiment. The experiment results showed that the optimal fermentation parameter consisted of initial pH of 8,30℃of fermentation temperature, 2% of inoculation amount,40% of medium loadage,120r/min of rotation, and 8d of fermentation time. The improved culture medium was composed of potato 200g/L, glucose 15 g/L, beef extract 20 g/L, NaCl 2 g/L, ZnSO4 0.08 g/L.CJ7 strain was fermented in large amount under the optimal fermentation conditions. Fermentation broth was separated and purified by macroporous resin, silica gel column chromatography and preparative HPLC. Two compounds were obtained from the bioactive part, which were named crystal A and crystal B.Crystal A and crystal B were screened on antimicrobial activity against 11 strains instruction bacteria and 1 strain instruction fungi by filter paper method. It was showed that crystal A had no antimicrobial activity against these strains while crystal B had antimicrobial activity against 9 strains. Crystal A and crystal B were evaluated on antitumor activity against human liver cancer HepG2 cell by MTT assay so as to further study their bioactivity. Crystal A and crystal B had not evident inhibit effect on HepG20 cell.The structures of two compounds were analyzed by means of MS and 1H-NMR. Crystal A could be a kind of compound in unsaturated fatty acids; Crystal B could be 2, 3-dihydroxypropyl palmitate.In this study the secondary metabolites of Acanthopanax senticosus endophytic fungi CJ7 were separated and purified, and compounds isolated from the secondary metabolites were assessed on antimicrobial activity and antitumor activity, which can provide important reference to the research of endophytic fungi and the development of new medicine.