Fermentation Of Fungal Endophyte Mutant M7226and Purification Of Its Secondary Metabolites

Objective:This study was based on the mutant M7226, which was obtained from the EZG0807of rhizoma zedoary. The fermentation condition was optimized by response surface method and the fermentation of M7226was began to mass produced. The appropriate separation method was designed to obtain the active substances by studying the properties of fermentation broth.Method:The Box-Behnken Design was used for the best fermentation conditions of strain M7226by the determined the influence factors and levels of this factors, then fermentation of M7226was conducted. In order to simplify the sepatation process, stability of fermentation broth on pH and temperature was tested by antibacterial experiment, fermentation broth was extracted by petroleum ether, chloroform, n-butanol successively and then tested by antibacterial experiment and reagent chromogenic experiments. Finally, Metabolites of strain M7226were separated by silica gel column chromatography, Sephadex LH20column chromatography, macroporous resin column chromatography and ODS column chromatography to obtained the pure compounds, which were identified by UV, IR, MS,1H and13C NMR.Result:1. The optimal fermentation condition of strain M7226was determined as pH6.8,29℃of fermentation temperature and10d of fermentation time by the BBD experiments.2. The metabolites of M7226were sensitive to pH but not to heat; The chloroform and n-butanol N-butyl extracts have inhibition effect on four indicator bacterias but petroleum ether extract don’t have.3. The fermentation of M7226was separated by silica column chromatography, Sephadex LH20column chromatography, macroporous resin D101column chromatography, ODS column chromatography, et. Five pure compounds were obtained and identified as curcumin, cinnamic acid,1,4-2hydroxy anthraquinone, gibberellic acid and kaempferol.