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標題: | 台灣紫芝生物活性代謝產物之醱酵生產與功能評估 Production and functional evaluation of bioactive metabolites using submerged culture of Ganoderma formosanum |
作者: | Ming-Hsu Chen 陳明煦 |
指導教授: | 劉文雄(Wen-Hsiung Liu) |
關鍵字: | 靈芝,台灣紫芝,醱酵,液態培養,生物活性代謝產物,細胞凋亡, Ganoderma,Ganoderma formosanum,fermentation,submerged culture,bioactive metabolites,apoptosis, |
出版年 : | 2006 |
學位: | 碩士 |
摘要: | 台灣紫芝為台灣地區特有之靈芝品種,目前關於此菌株之文獻及研究報告相當稀少。本研究之目的為建立台灣紫芝於五公升醱酵槽中之最適液態醱酵條件,評估醱酵產物之抗腫瘤活性,及純化最具有生物活性之成分。藉由台灣紫芝在 500 mL Hinton 氏三角瓶培養之結果,選出具有生產較多菌絲體及生物活性代謝物潛力之 G. formosanum ATCC76538 做為生產菌株。在五公升醱酵槽中探討生產菌絲體及胞外多醣之最適接種量、攪拌速率、通氣量之結果為: 最適接種量 10%, 最適攪拌速率 150 rpm, 最適通氣量 1 vvm。醱酵濾液以乙酸乙酯進行萃取,所得之萃取物在細胞測試中對肺纖維母細胞 (MRC-5)、肝癌細胞 (Hep3B、HepG2)、子宮頸癌細胞 (HeLa)、肺癌細胞 (CL1-1) 均有明顯抑制生長之效果,進一步之研究發現紫芝萃取物或許能經由粒腺體依賴性細胞凋亡路徑誘發 Hep3B 細胞進行細胞凋亡。由醱酵液萃取物純化所得之活性物質 F4652,在劑量增加下可明顯對 Hep3B 細胞之生長產生抑制。利用固相萃取法及半製備式管柱加速 F4652 之純化流程,並以 GC-MS 分析其分子量。 Ganoderma fromosanum is a unique Ganoderma sp. in Tawain. Until now, there have been just a few literatures and studies on this strain. The purpose of this study is to optimize the fermentation conditions of G. formosanum in a 5-L fermentor, to evaluate the antitumor effect of fermentation products and to purify the the most biological active component. We chose G. formosanum ATCC76538 as the production strain because it showed more potential on mycelia growth and bioactive metabolites production in a 500 mL Hinton flask cultivation. The effect of inoculum size, agitation speed, aeration rate on production of biomass and extracellular polysaccharide were carried out in a 5-L fermentor. As it was found that the optimal cultivation conditions were as follows: inoculum size, 10%; agitation speed, 150 rpm; aeration rate, 1vvm. The fermentation filtrate was extracted with ethylacetate and the extract showed significant growth inhibition to human lung fibroblast cell (MRC-5)、hepatocellular carcinoma cell (Hep3B)、hepatoblastoma cell (HepG2)、cervix epithelial carcinoma cell (HeLa) and lung carcinoma cell (CL1-1). Further investigation showed the extract might induce apoptosis in Hep3B cell through mitochondria dependent pathway. An active compound F4652 was purified from filtrate extract, and showed growth inhibiton against Hep3B cells in a dose-dependent manner. We used solid-phase extraction and semi-preparative column to fasten the purification process and used GC-MS to analyze the molecular weight of F4652. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/32738 |
全文授權: | 有償授權 |
顯示於系所單位: | 微生物學科所 |
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