Solid Fermentation Production Of Ergosterol With Penicillium Chrysogenum

Ergosterol,the main raw material to production of vitamin D₂,is an important pharmaceutical intermediate.And it is mainly located on the fungal cell membrane, which can be used for characterization fungal biomass,it has been becoming one of the most important tumor markers of fungal biomass of the fungal cell membrane.In this paper,solid fermentation production of ergosterol with Penicillium chrysogenum was studied.Qualitative was analysised by Thin-layer Chromatograph(TLC),Ultraviolet Spectrameter(UV) and HPLC.The results show that the crude extract of Penicillium chrysogenum solid fermentation material contains Ergosterol,It had been established that a determination of ergosterol quantitative method which is TLC-UV,and corresponding calculate relationship is established.It was studied that the specification of saponify and extracting ergosterol from Penicillium chrysogenum.Alcohol,ethanol as well as sodium hydroxide were used as saponification menstruum,respectively.The optimal condition of ergosterol extraction was concluded by means of orthogonal design test.Rate of fermentation product vs. saponification menstruum(m/v) was 1:5,and rate of alcohol,ethanol and sodium hydroxid(15%)(v/v) was 2:1:2.Saponification temperature was 90℃.Saponification time was 90min.Fresh Penicillium chrysogenum mycelium can be obtained by flat culture then freeze the substrate.Validate the linearity relation of the content of ergosterol and biomass and the effect of culture time and substrate contain.We get the conclusion that mycelium biomass has a good linear relation with the content of Ergosterol.Culture time and substrate contain little effect to content of ergosterol.The relation(y= 4.354 10^-3x) between fungus myceliurn(x/g) and the ergosterol content(y/g) was established.The optilmal fermentation composition and culture condition were studied. According to single-factor experiment,it was found that addition of glucose, carbamide as well as monosodium orthophosphate to solid substrate containg both wheat bran and bean bran,accumulation of ergosterol was enhanced efficiently.An ideal composition of the solid substrate for fermentation included wheat bran 8g,bean bran 2g,glucose 0.8g,carbamide 0.06g and KH₂PO₄ 0.01g.The optimal culture medium was as follows:water rate is 1.0mL·g^-1,fermentation time is 5.5 days.Under the optimum condition,the output of ergosterol may reach 2.953mg/g(dry culture medium).Studies the isolation and purification Ergosterol from the Penicillium chrysogenum crude extract by silica-gel column chromatography.Eluate used for the petroleum ether-diethyl ether mixed solvent system.volume ratio of 5:2.Qualitative analysis was made by UV and HPLC.The result is satisfactory.The purity of ergosterol is 85.6%.