費氏麴菌二次代謝物之培養最適化條件與生物活性研究

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Title:	費氏麴菌二次代謝物之培養最適化條件與生物活性研究 Studies on the optimal growth condition and biological activities of secondary metabolites from Neosartorya fischeri
Authors:	趙昱揚
Contributors:	藥物科技研究所李冠漢
Keywords:	生物轉化費氏麴菌抗氧化二次代謝物大黃黃柏黃芩 Radix Scutellariae Cortex phellodendri antioxidant Radix et Rhizoma Rhei secondary metabolites biotransformation Neosartorya fischeri
Date:	2012
Issue Date:	2014-03-07 11:06:21 (UTC+8)
Abstract:	本研究目的為探討費氏麴菌二次代謝產物生合成的最適化條件與其生物活性。依照標準培養步驟配置YMG培養基（酸鹼值5.5）在37℃下培養費氏麴菌，培養二週。以萃取方式每天提取費氏麴菌培養基中二次代謝物，並以高效能液相層析儀分析每天變化的情形。在層析圖譜上滯留時間為22.5分鐘有主要吸收峰，藉由質譜與核磁共振譜鑑定其結構為acetylaszonalenin，將acetylaszonalenin視為在費氏麴菌二次代謝物的指標成分。進一步利用環境因子包括：酸鹼值、溫度、碳源、氮源等的改變，觀察費氏麴菌二次代謝物產量的變化。應用生物轉化的技術，於培養基中添加中藥大黃、黃柏、黃芩等水萃取物，以細胞存活率分析和自由基清除率試驗評估其生物轉化前與轉化後的生物活性。結果顯示，費氏麴菌在起始 pH 4.5、溫度40℃條件下，培養天數至第7天時能提高acetylaszonalenin 0.61 mg的產量。並分別在第11天與第14天出現較高產能。碳源與氮源濃度減少為0.5倍與缺乏條件下，造成acetylaszonalenin的產量降低。氮源改變為 meat peptone 培養時產量增加1.29 mg。顯示在起始pH 4.5、溫度40℃、氮源改變為 meat peptone 的條件培養比原條件起始 pH 5.5、溫度37℃，提高3.34倍的產量。生物活性結果顯示，添加黃柏水萃物，明顯提高N. fischeri抗氧化的能力；但大黃水萃物的添加則在生物轉化後所得的二次代謝物，其抗氧化能力有逐漸降低的趨勢。 The aim of this study is to investigate the optimal growth condition and biological activities of secondary metabolites from Neosartorya fischeri. According to protocol, the Neosartoya fischeri deployed YMG medium (pH 5.5) and cultured under 37 °C for two weeks. The total amount of secondary metabolites from N. fischeri was extracted and collected every day and analyzed by using high performance liquid chromatography (HPLC). We observed a peak appeared in HPLC chromatogram at Tr=22.5. After purification and structure elucidation by MS, 1D and 2D NMR, we identified this compound as acetylaszonalein. Therefore, we use acetylaszonalenin as the reference compound for the subsequent secondary metabolites studies of N. fischeri. The environmental factors for growing N. fischeri, such as pH, temperature, carbon sources, nitrogen sources had been changed to investigate the changes of secondary metabolite profiles. Futhermore, we utilized the biotransformation technology to co-culture with the extracts of Chinese herbal medicines, such as Radix et Rhizoma Rhei, Cortex phellodendri, and Radix Scutellariae and to evaluate the preliminary biological activity by MTT and DPPH assay. We concluded that the production of secondary metabolites, acetylaszonalenin, was increased by using pH 4.5, 40℃ and meat peptone nitrogen source as growth condition. When the starvation studies proceed, the production of acetylaszonalenin biosynthesis decreased. The biotransformation of N. fischeri with Cortex phellodendri water extract significantly improved the antioxidant activity of N. fischeri.
Relation:	校內一年後公開，校外永不公開，學年度：100，87頁
Appears in Collections:	[Dept. of Pharmacy] Dissertations and Theses

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