Study On Microbial Fermentation Modification Of Small Hundred And Kunlun

The vital movements of microbes are very vigorous, the majority of them could make a full use of the effective ingredients in culture mediums for the growth and metabolism. In recent years, with the continuous development of modernization of Traditional Chinese Medicine (TCM), the technology that using the microbial decomposition abilities into the Chinese herbal medicine processing has been attracted extensive attentions of the experts all over the world. We can say that the microbial fermentation and modification of TCM, is bound to pave a new road for the development of TCM. This paper studied the fermentation and modification of traditional chinese medicine Asparagus filicinus and Coreopsis tinctoria with ten different pathogenic fungi strains. After the chemical composition and activity analysis of fermented crude extracts, we have had an intensive understanding of the modified mechanism with microbial fermentation. This thesis was composed of three chapters:In chapter 1, the solid fermentation of A. filicinus was investigated. Some chemical composition changes were detected after fermentation with Fusarium oxysporum by use of TLC and HPLC. After separation and analysis of the compound, rubrosterone was determined, and 20-hydroxyecdysone was confirmed as the precursor compound of this biotransformation, the conversion ratio between them was close to 100%. Then, the research of different fermentation conditions showed that 20-hydroxyecdysone could be transformed into rubrosterone in a high yield only under the condition of solid fermentation. However,β-sitosterol and β-daucosterol couldn’t be transformed to corresponding androsterone compounds by F. oxysporum under solid fermentation. In addition, the chemical characterization of solid fermented A. filicinus with NMR showed that F. oxysporum fermentation could greatly reduce the sugar content of A. filicinus, which was more convenient for the subsequent extraction and separation. Finally, different acvities (anti-tumor activity, antioxidant activity in vitro, anti-pathogenic activity) of fermented crude extracts and compounds were also screened.The results showed that microbial solid fermentation could greatly improve the antioxidant activity of A. filicinus, and the crude extracts of solid fermented A. filicinus had a great improvement on the anti-pathogenic activity of Bacillus subtilis and Escherichia coli. This research provided an innovative approach to accumulate rubrosterone with mild reaction conditions and high conversion rates, at the same time, provided an idea for the improvement of antioxidant activities and anti-pathogenic activities of Chinese medicinal materials.In chapter 2, the solid fermentation of Coreopsis tinctoria was studied. The total phenolic content (TPC) and total flavonoid content (TFC) of the crude extracts of C. tinctoria fermented with different microbial indicated that the TPC and TFC were increased obviously with the fermentation of Aspergillus niger and Stagonosporopsis cucurbitacearum. The study of the antioxidant activities in vitro of fermented C. tinctoria in different methods (DPPH radical-scavenging activities, ABTS radical-scavenging activities, Ferric reducing-antioxidant power) was carried out. The antioxidant activities of fermented C. tinctoria with A. niger and S. cucurbitacearum revealed an increasing trend, and showed a good correlation between TPC and TFC, which illuminated that phenolics and flavonoids played an important role in the antioxidant activities of these extracts. In addition, UV、NMR、IR、HPLC were also used for the characterization of chemical changes of nonfermented C. tinctoria, fermented C. tinctoria with S. cucurbitacearum and A. niger. This study provided a reference for the modification research on microbial fermentation of C. tinctoria, and has been heralded as the alternative strategy for the Chinese herbal medicine processing.In chapter 3, the transformation reaction mechanism of steroid with microbial and the application of transformation technology in steroid compounds were reviewed in recent years.