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研究生:陳振輝
研究生(外文):Chen-Hui Chen
論文名稱:新型抗氧化劑的分離、純化和活性分析
論文名稱(外文):Isolation, purification and characterization of novel antioxidants
指導教授:蔡英傑蔡英傑引用關係
指導教授(外文):Ying-Chieh Tsai
學位類別:碩士
校院名稱:國立陽明大學
系所名稱:生物化學研究所
學門:生命科學學門
學類:生物化學學類
論文種類:學術論文
論文出版年:2001
畢業學年度:89
語文別:中文
中文關鍵詞:抗氧化劑低密度脂蛋白
外文關鍵詞:antioxidantPenicillium citrinumgentisic acidscopoletinlow density lipoprotein
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本篇論文描述了Hui-1、gentisic acid、scopoletin的抗氧化特質。 我們從Fujisawa Co.提供的3562株菌的萃取液中, 篩選得到一株具有高抗氧化活性的真菌: Penicillium citrinum (F5), 在大量培養後, 分離得到兩個具抗氧化活性的物質, Hui-1和Hui-2, 經結構鑑定後確定Hui-2為2,5-dihydroxybenzoic acid, 即gentisic acid。 scopoletin的抗氧化性, 也同時一起作探討比較。 scopoletin是實驗室過去從中藥青楓藤中純化到的物質。
從初步活性分析的結果看來, Hui-1和gentisic acid都有著很強的還原力, 還原自由基DPPH的能力和ascorbate及trolox相當。 在LDL氧化實驗中, 100 mM濃度下, 兩者都可以完全保護LDL免於AAPH引起的氧化傷害。 scopoletin則特別具有捕捉superoxide anion的能力。 因為ROS造成的傷害和許多疾病都有關係, 進一步深入的研究自然有其價值。

This paper describes the antioxidant properties of Hui-1, gentisic acid, scopoletin. After screening 3562 strains of microbial extracts from Fujisawa Co., one strain of Penicillium citrinum (F5) with high antioxidant activity was found. Two potent antioxidative compounds, Hui-1 and Hui-2, were isolated from the culture broth of this strain. Hui-2 was identified as 2,5-dihydroxybenzoic acid (gentisic acid). Besides, scopoletin, isolated from Sinomonium acutum (a traditional Chinese herb) in our lab before, was investigated at the same time.
Hui-1 and gentisic acid were as effective as ascorbic acid and trolox in reducing the stable free radical DPPH. They also inhibited oxidation of LDL induced by peroxyl radical (AAPH) completely at 100 mM. Scopoletin scavenged superoxide anion specifically without inhibiting xanthine oxidase but was inactive in other assays. As reactive oxygen species caused damages are related to various diseases, the further research of above novel antioxidants are quite justified.

中文摘要 ---------------------------------------------------- 3
英文摘要 ---------------------------------------------------- 4
縮寫表 ------------------------------------------------------- 5
第一章 緒論 ---------------------------------------------- 6
第一節 抗氧化劑的定義 ------------------------------------------------- 6
第二節 抗氧化劑的評估 ------------------------------------------------- 6
第三節 ROS (reactive oxygen species) 的成員 ----------------------- 9
第四節 抗氧化活性的評估方式 --------------------------------------- 12
第五節 ROS造成的疾病 ------------------------------------------------ 13
第六節 研究動機與目的 ------------------------------------------------ 18
第二章 材料與方法 ------------------------------------ 19
第一節 材料 --------------------------------------------------------------- 19
第二節 方法 --------------------------------------------------------------- 21
第三章 結果 --------------------------------------------- 29
第一節 抗氧化活性菌株的篩選 --------------------------------------- 29
第二節 Penicillium citrinum (F5) 的活性物質純化 ---------------- 36
第三節 Hui-2的結構鑑定結果 ---------------------------------------- 42
第四節 Hui-1及gentisic acid的抗氧化活性分析 ------------------- 42
第五節 scopoletin的抗氧化活性分析 -------------------------------- 47
第四章 討論 --------------------------------------------- 52
第一節 篩選實驗的設計 ------------------------------------------------ 52
第二節 Hui-1和gentisic acid的相關研究 ---------------------------- 52
第三節 scopoletin的相關研究 ----------------------------------------- 53
第四節 實驗總結 --------------------------------------------------------- 53
第五章 參考文獻 --------------------------------------- 56
附錄一、F5菌種鑑定報告 ------------------------------ 60
附錄二、Hui-2的結構鑑定資料 ----------------------- 63

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