Studies On The Anticancer Active Components Of The Endophytic Fungi Fermentation Broth

Endophytic fungi FSN002 was firstly isolated from the bark of the Chinese catalpa and defined as the trichoderma longibrachiatum. We adopted MTT method to do the anticancer activity detection of the fermentation broth of FSN002. The results showed that the fermentation broth could inhibit the growth of liver cancer cell line HepG2 effectively and also has strong inhibition effect on lung cancer line SPC-A-1 and gastric cancer line SGC. At the same time, the fermentation broth reflects little effect on normal liver cell line HL-7702. In a word, the fermentation broth of FSN002 shows good anticancer effect.In this paper, firstly we studied the correlation between the pellet shape and the anticancer activity of endophyte FSN002, and we found that different pellet shape of FSN002 has different level of anticancer activity and different the metabolic spectrum. In order to improve the stability of the fermentation process and gain the stable anticancer activity of the fermentation broth, we selected monoclonal strains through the way of single-cell spore cultivation and got series of monoclonal strains with different anticancer activity and pellet forms. And three monoclonal strains were found with high and stable anticancer activity.Secondly we learned the relationship between the growth of fungi and the production of anticancer substance with the fuzzy mathematical model, it showed the fact that growth phrase and production of anticancer material process separated, i.e., growth and production phase separation.Then we systematically studied the stability of the fermentation broth in different processing conditions like temperature, organic solvents, acid and alkali to lay the foundation for the separation of anticancer activity component.We found that the temperature that lower than 100℃ and differnent organic solvents(methanol, ethanol, chloroform, dichloromethane, benzene, cyclohexane, ethyl acetate, petroleum ether didn’t show any effect on the stability of the fermentation broth. And the fermentation broth can keep its stability under netural or partially alkaline conditionsFinally we took the HPLC method to study the metabolic spectrum of the endophytic fungi FSN002. Combineing with the anticancer activity test and statistical analysis, we found that there existed positive correlation between the amount of some yellow substance calledXiuLingsu in the fermentation broth and the anticancer activity, and Xiu Lingsu is the candidate of active anticancer properties. The results also showed that the total yellow substance in the fermentation consisted of the medium hydrophobic yellow anticancer activity component and high hydrophobic yellow impurities. Further, we confirmed the element formula(C28H3508) of the anticancer activity component-XiuLingsu through the super high performance liquid chromatography and mass spectrometry analysis and made a solid foundation for the later separation of anticancer activity component.