Identification of the Diterpenoids Produced by Endophytic Fungus of Torreya fargesii and Its Inhibitory Effect on Hela Cells

摘要

[Objectives] The purpose of this study is to dissociate endophytic fungus producing diterpenoids from Torreya fargesii tissue and examines its inhibiting effect on tumor cells. [Methods]Plant endophytes were isolated and purified to study their resistance to Gram-positive( G+) and Gram-negative bacteria( G-). High performance liquid chromatography( HPLC) was used for analysis of the retention time,relative peak area and percentage content of its metabolite. By liquid chromatography-mass spectrometry( HPLC-MS),the material characteristic of the ion pair information of the metabolites was measured. The bacterial strain was also classified. [Results] The results showed that the secondary metabolites produced by the strain BP6 T3 possessed double resistance to G+and G-bacteria. The strain was identified as Penicillium sp by preliminary classification. Through HPLC analysis,the retention time of fermentation extracts was 12. 8 min with almost the same as the standard of taxol. According to the chromatograph,the relative peak area was 12 887. 11,the average relative percentage was about 15. 8%,and the content of taxol analogs in fermentation broth reached 16. 59 mg/L. The material characteristic of the formation of ion fragments of taxane analogues in metabolic extracts was identical to that of the taxol standard determined by HPLC-MS. It can be initially determined that strain BP6 T3 can produce taxane compounds. Taxol substance produced by this strain had obvious inhibitory effect on Hela cells with the concentration increasing. Different precursors had a significant effect on the production of paclitaxel metabolites in this strain. L-phenylalanine was used as the precursor and the yield increased most,with an increase rate of 425. 7%. [Conclusions] The strain is expected to be used for mass production in antitumor drug taxol.