Study On The Structure Of Sedative And Hypnotic Bioactives Of Paecilomyces Gunnii

Paecilomyces gunnii, the anamorph of Cordyceps gunnii, is a fungus with high medicative value and promising utilization perspectives. The active components in the fermentation mycelia of the strain which isolated from Cordyceps gunnii were analyzed, the most active strains were screened out used as subsequent experimental material. The mycelium powder were extracted by ethanol firstly, then extracted by petroleum ether, ethyl acetate, butanol and methanol. The most significant effective extraction with sedative and hypnogenesis were screened by rat experiments. Based on the screening, the extraction were further separated and purified by through silica gel column. The significant sedative compounds were detected through a hypogenesis and calm model. UV spectroscopy, liquid mass spectrometry and nuclear magnetic resonance analysis were used to determine the structure of the material. The specific contents include isolation of high yield and quality strains, separation of the active ingredients and structure identification of active compounds.The analysis of effective components in the submerged liquid fermentation mycelia of Paecilomyces gunnii were used to screen out the high yield, rich secondary metabolites and rapid growing strain as following exprimental material.Hypnogenesis and calmness activity of the four different extractives from the fermentation were detected through a hypnogenesis and calm model. The results showed that butanol extract had shtronger activity than the other extractives.Based on the above screening, the butanol extract was isolated and purified with HPLC, LC-MS, and column chromatography, and six components were obtained, which were named from Ⅰ to Ⅵ. Hypnogenesis and calmness activity of the six different components from butanol extract were detected through a hypnogenesis and calm model. The results showed that component Ⅰ had stronger activity than the other components.The component Ⅰ was isolated and purified with HPLC, and four compounds were obtained, which were respectively named pg-1, pg-2,pg-3and pg-4. The stronger activity compound, pg-3, was screened out through animal hypnogenesis and sedative model. ESI and HPLC-MS analyses revealed the molecular formulas of pg-3as C20H32N6O4S1and its molecular weight was452.23003. The structure of compound C20H32N6O4S1was detected by NMR analyses. Animal expriment showed that the purified compounds had strong hypnogenesis and calm activities with values of51.59%and41.06%. It suggested that the separation and purification of the bioactive components from the extract was successful.The culture characteristics and colony morphology of ten isolated strains of Paecilomyces gunnii were observed. Besides, their sporulation yield and content of major components such as protein, polysaccharide, mannitol, ergosterol, adenosine were measured. The result showed that different separated strains had different morphological characteristics and there was certain relationship between phenotypic diversity and main ingredients.Liquid culture conditions and extraction process of ergosterol of Paecilomyces gunnii were optimized using biomass and ergosterol content as index. The result of orthogonal test indicated that the optimal liquid medium is:sucrose4%, bran2%, KH2PO40.025%, NaCl0.05%. Response surface experiment demonstrated that the optimal extract solvent concentration was80%ethanol, the ratio of solvent to raw material was200:1and extract time was65min. Results of the this study could be a reference for development of Paecilomyces gunnii.