Optimization Of Fermentation Conditions For The Mycelial Biomass Medicinal Mushroom Phellinus Pini And Its Antineoplastic Activity Analysis

Phellinus pini, a basidiomycete fungi belonging to the genus of Fomes and the fruit body called "Huangzhi" in China, a traditional medicinal mushroom have been used to treat various tumor diseases in Asian, such as esophagus, gastric, lung breast, colon and rectum cancer. However, the main limitation of fungus resource in nature and increasing in search for substitutes of the natural products of fungus. Fermentation technology has played an important role to discover new bioactive metabolite. Therefore, optimization of culture medium for P. pini is necessary to search for natural products, and investigate antineoplastic mechanisms.1. In this study, statistical designed experiments (one-factor-at-a-time, Plackett-Breman, steepest ascent design, CCRD) were applied to optimize the culture medium for P. pini mycelia dry weight. Finally, the optimum medium composition was identified as followed:85g L-1soluble starch,60g L-1glucose,1g L-1CaCl2,0.8g L-1MnSO4,15g L-1beef extract,25g L-1TSB,0.5g L-1KH2PO4and0.5g L-1MgSO4·7H2O, the maximum biomass dry weight was verified28.95g L-1as increased by7-10folds compared to original medium culture.2. Biological activity substances were also investigated, extracts both from mycelia and culture broth tested on growth human gastric cancer cell line SGC-7901in vitro. Cell viability assay by MTT method displayed that intensely antineoplastic activity was observed after petroleum ether extract treated, with an IC50value of6.26ugml-1at24h. 3. Objective To study the chemical constituents if Medicinal mushroom Phellinus pini. Methods Phellinus pini was cultured in liquid medium, its secondary metabolites in culture broth were extracted by Ethyl Acetate. The extract was subjected to chromatography on silica, Sephhadex LH-20column and HPLC. The compounds were identified on the basis of their physicochemical and spectroscopic methods. Results Six compounds were isolated and their structures were educidated:4-vinylphenol, Adenine5’-Deoxy-5’-methylthiopentoside, squalene, Drimenol, Drimendiol and p-Vinylphenyl β-D-Glucopyranoside. Conclusion All the compounds except for4-vinylphenol are isolated from the fungi for the first time.4. MTT methods was used to analysis antineoplastic activity of compounds in growth of SGC-7901cells in vitro, the GES cells was also been tested. It was showed cell growth promoting activity after the compounds treated24h, this result in accordance with the research results had been reported. It is suggested the P. pini was a antineoplastic Medicinal Material for further study.Conclusion, these works were contributed to solve the main limitation of fungus resource in nature and increasing in search for substitutes of the natural products of fungus, and analysis the antitumor activity of P. pini. It could be useful for development of P. pini of medicinal mushroom.