| Summary: | 碩士 === 國立臺東大學 === 生命科學系碩士班 === 100 === Antrodia camphorata is a unique fungus in Taiwan, the fruit body of Antrodia camphorata often used as liver disease treatment. But the fruit body of Antrodia camphorata is difficult to be obtained, and also expensive, now submerged fermentation is usually used to culture the mycelia of Antrodia camphorate. The deep ocean water (DOW) contains rich metals and trace elements (Ca2+, Mg2+, PO42-, etc.) promoting the yeast growth or functional activity of Aspergillus. In this study, the fermentation study was divided into four phases; first we used different proportions of DOW (0%, 25%, 50%, 75%, and 100%) for the fermentation of Antrodia camphorata, then measure the antioxidative ability and component of Antrodia camphorata. The assessment of the targets included: biomass, free radical scavenging test, reductive ability determination, triterpenoids analysis, polysaccharides quantification, total polyphenols quantification, flavonoids quantification, etc. Second we added the extra major ions at a equal concentration with the (Mg2+, Na+, Ca2+, Zn2+, Fe2+, K+) of DOW into ultra-pure water (UPW) medium for the culture of Antrodia camphorata in order to understand the production of the functional metabolites of Antrodia camphorata, and select the major ions stimulation the growth and metabolite production of Antrodia camphorata. Third the ions and nitrogen source (yeast extract) with increasing concentrations into was added DOW medium in order to select the optimal concentration of target ions and yeast extract for the fermentation of Antrodia camphorata. Fourth the concentration of target ions and yeast extract were selected as the three factors of response surface methodology (RSM) to investigate the optimal condition for the fermentation of Antrodia camphorata.On the result increasing the proportion of DOW water increased the biomass triterpenoids, polysaccharides, total polyphenols, and total flavoids. Antioxidative test showed that Antrodia camphorate fermented using 100% DOW has better antioxidative ability, therefore, DOW effectively enhance the production of the functional metabolites of Antrodia camphorate.Furthemore, Mg2+ and Zn2+ had more effect on enhancing the mycelia growth. The third stage results show that DOW medium including Mg2+ and yeast extract was able to enhance biomass, intracellular polysaccharide and extracellular polysaccharide content of Antrodia camphorate with dose dependent-response. Therefore, Mg2+, yeast extract and glucose were selected as the three factors of RSM for the study of optimal condition. The statistic regressions of the target effectors had high R2 value. The optimum conditions was selected according to the overlapped plot, which is 1.65 g/L Mg2+, 15.85 g/L yeast extract, and 4% glucose. In the evaluation for the prevention from Liver crossis, Antrodia camphorate-fermentated product (DOW-AC-1X) cultured by DOW prevented the weight loss induced by TAA, and had more effect on inhibiting the MDA and ROS generation than one dosage of ROW-AC. The western blot experiments show that the iNOS protein expressing of DOW-AC-1X and DOW-AC-2X are lower than that of ROW-AC-1X. The histologic H&E stain and collagen stain showed that DOW-AC-1X has a better ability to prevent the liver damage and crossis, and DOW-AC-2X has a more significant effect. The results showed that using DOW as culture water functional metabolite production increase the ability of the fermentation and the liver protection of Antrodia camphorate under the submerged fermentation.
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