Studies On Submerged Fermentation And Triterpenoids Of Ganoderma Applanatum

With the development and progress of human medicine, more and more attention has been paid on the medicinal value of fungi, especially Ganoderma, mostly of them can be used as medicine and will be good for health. In recently years, chemical composition analysis, extraction, separation and studies on biological activities of Ganoderma were getting attention. Triterpene compound widely exists in Ganoderma with the kind of anti-cancer activity component. Submerged fermentation,X-ray spectrometry scanning were used to optimizing the Liquid medium to improve content of terpene compounds in mycelia and contrast to that of fruit bodies of Ganoderma applanatum, so that it may provid the theory basis for development and utilization on resources of Ganoderma applanatum.Efficiency of microwave extraction, ultrasonic method, saxhlet extraction and immersing extraction of triterpene compounds were compared to get the best extraction method. The result shows that microwave extraction is good method. Then the optimum conditions for microwave extraction of triterpenoids was optimized. It chooses extraction time, extraction temperature, liquid material as optimization factors of the single-factor test, and choose triterpenoid content of Ganoderma applanatum as the optimization condition, and according to the Box-Behnken central composite experimental design principle, and deal with the model data using the Design-expert7.1.6software program to establish the response surface method of mathematical model data processing, so it obtains the optimum conditions of microwave extraction of triterpenoids:Liquid materials of extraction solvent and the sample is20ml·g-1, extracted for15min under constant temperature75℃.It is the first time to use the method for extraction of triterpenoid of Ganoderma applanatum. Advantages is less solvent, short time, improved extraction efficiency, and easy operation.Submerged fermentation was conducted to get a large quantity mycelia of Ganoderma applanatum to compare the triterpene content between fruit bodies and mycelia of Ganoderma applanatum, and may provide theoretical basis for gaining triterpene activity material from mycelia. Orthogonal test was used to get the best proportion of carbon and nitrogen sources and to optimize medium of submerged fermentation to obtain triterpene of Ganoderma applanatum. First a single factor experiment was done, then considering the preparation costs of the culture medium, and chooses sugar, brown sugar, potatoes as different carbon nitrogen source, based on L9(34), and chooses mycelium yield and total triterpenoid content as a scalar, proceeding optimization of culture medium. Elements in fruit bodies and mycelia of Ganoderma applanatum was determined by X-ray spectrometry scanning, and the results so that it have slightly different elements in Mycelia and fruiting bodies, providing more scientific basis for adjusting liquid culturate formula of mycelia.The results show that tht best carbon and nitrogen sources formula of Ganoderma applanatum submerged fermentation medium:sucrose12.5g· L-1, brown sugar,12.5g·L-1, potato25g·L-1.Microwave extraction, Saxhlet extraction, Column chromatography, thin layer chromatography (TLC), HPLC, GC-MS were used on extraction and separation of chemical constitution of Ganoderma applanatum. It shows a steroid compound was separate fromchloroform extraction.38compounds were identified by GC-MS analysis from fat soluble constituents of Ganoderma applanatum, They are alkanes, ester, acid, amine,Ergosterol aldehyde, formate, Butylated Hydroxytoluene and5,6-Dihydroergosterol besides etc. It confirm preliminary that it get a compound from chloroform extraction parts, identified as stigmasterol.The antibacterial activity test of extraction of fruit badies and mycelium of G. applanatum were carried out.The result shows that:friuting badies and mycelium of G applanatum have different levels of bacteriostasis on the three instructions bacteria. Analyse by antibacterial rate, the inhibitory effect of mycelium extraction is significantly stronger than the fruit bodies about chloroform extraction, showing chloroform extraction of mycelia may have a higher content of antibacterial activity material; the inhibitory effect of the fruit bodies extraction is significantly stronger than mycelium of ethyl acetate extraction, showing ethyl acetate extraction of fruit badies may have a higher content of antibacterial activity material.