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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 羅翊禎 | zh_TW |
| dc.contributor.author | 林欣誼 | zh_TW |
| dc.contributor.author | Hsin-I Lin | en |
| dc.date.accessioned | 2021-07-11T15:14:32Z | - |
| dc.date.available | 2024-08-07 | - |
| dc.date.copyright | 2019-08-07 | - |
| dc.date.issued | 2019 | - |
| dc.date.submitted | 2002-01-01 | - |
| dc.identifier.citation | Borovikova, D.; Teparić, R.; Mrša, V.; Rapoport, A., Anhydrobiosis in yeast: cell wall mannoproteins are important for yeast Saccharomyces cerevisiae resistance to dehydration. Yeast 2016, 33 (8), 347-353.
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/78720 | - |
| dc.description.abstract | 苦瓜為亞洲常見之瓜果類蔬菜，具有許多功效性成分，使之成為歷史悠久之藥用植物。皂苷為苦瓜中重要的活性成分之一，其中，3β, 7β, 25-trihydroxycucurbita-5, 23 (E)-dien-19-al (THC) 被發現對誘導引發有醣尿病之小鼠具有降血醣之功效。因此本實驗希望透過酵母菌發酵山苦瓜以提高具降血醣功效成分3β,7β,25-trihydroxycucurbita-5,23(E)-dien-19-al之含量。實驗先期透過TLC及HPLC-MS/MS分析篩選具皂苷水解能力之菌株，結果發現酵母菌 YCL-L-019 及 YCL-L-020 具轉換 momordicoside L及 momordicoside K 之能力，同時可顯著提升 THC 之含量，並且YCL-L-020之轉換速度比YCL-L-019快。透過滲透壓、剛果紅及 zymolyase 敏感性試驗觀察到YCL-L-020對於滲透壓及剛果紅有較高之敏感性，並於使用SDS-PAGE分析不同酵母菌株之胞外蛋白時亦觀察到YCL-L-020 相對於YCL-L-019有較為複雜之組成，可初步推測 YCL-L-019 及 YCL-L-020 細胞壁存在有差異，為導致其胞外蛋白組成較為複雜及轉換苦瓜皂苷速度較快速的可能原因之一。欲進一步探討是何種酵素作用造成苦瓜皂苷之轉換，本實驗透過分析 YCL-L-020 之胞外蛋白質體及資料庫搜尋推測可能之作用酵素，並以建構質體之方式使 S. cerevisiae 表現推測之酵素，且將表現有該酵素之菌株培養於含苦瓜甲醇萃取物之基質中，觀察該菌株轉換苦瓜皂苷之效果。最終於表現YCL-L-020 BGL1之菌株中觀察到轉換苦瓜皂苷之效果且轉換效果與YCL-L-020之轉換效果相似，後續可再針對該酵素之活性優化及應用進行探討。 | zh_TW |
| dc.description.abstract | Momordica charantia L., also known as bitter gourd, belongs to family Cucurbitaceae and has been used as herbal medicine for a long time in Asia and Africa. Previous studies indicated that saponins play important roles in the bioactivity of Momordica charantia. Particularly, 3β, 7β, 25-trihydroxycucurbita-5, 23 (E)-dien-19-al (THC) is specific saponin that has been reported to have hypoglycemic effects in the diabetes-induced male ddY mice. The aim of this study is to find a unique yeast can enhance the content of THC. In the analysis of thin layer chromatography and HPLC-MS/MS, we found that YCL-L-019 and YCL-L-020 are both able to transform momordicoside L into THC while YCL-L-020 convert more rapidly compared to YCL-L-019. By the test of osmotic sensitivity, congo red sensitivity, Zymolyase sensitivity, and SDS-PAGE analysis, we propose that the difference of cell wall between YCL-L-019 and YCL-L-020 might be a reason why YCL-L-020 have a higher speed in saponin transformation than YCL-L-019. To find out which enzyme is critical for YCL-L-020 to transform the saponins of Momordica charantia further, we use secretome analysis and database searching to screen candidates and clone these enzyme in plasmid and transform into Saccharomyces cerevisiae BY4741 mutants that can’t convert bitter melon saponin in previous experiment. Successfully, we found that the S. cerevisiae BY4741 expresses the BGL1 of YCL-L-020 can specifically convert momordicoside L into THC. | en |
| dc.description.provenance | Made available in DSpace on 2021-07-11T15:14:32Z (GMT). No. of bitstreams: 1 ntu-108-R06641017-1.pdf: 12841355 bytes, checksum: d06bf952e1c493808e4f7a6e7b32f9ce (MD5) Previous issue date: 2019 | en |
| dc.description.tableofcontents | 摘要 i
Abstract ii 目錄 iii 圖目錄 v 表目錄 vi 附錄目錄 vii 第一章、前言 1 第二章、文獻回顧 2 第一節、苦瓜 2 一、苦瓜簡介 2 二、苦瓜皂苷 3 三、皂苷分析方法 5 第二節、生物轉換 7 一、生物轉換具醣苷之苦味物質 7 二、三萜類皂苷之生物轉換 7 第三節、酵母菌細胞壁 9 一、幾丁質 9 二、葡聚醣 9 三、甘露醣蛋白 10 第四節、蛋白質體分析 11 第三章、研究目的與實驗架構 12 第四章、材料與方法 14 第一節、實驗材料 14 一、山苦瓜 14 二、酵母菌菌株 14 三、質體 14 四、引子 14 五、藥品與試劑 19 六、儀器設備 20 七、統計軟體 21 第二節、實驗方法 22 一、酵母菌生物轉換苦瓜皂苷萃取物 22 二、基本特性確認 24 三、外泌蛋白分析 25 四、作用酵素探討 28 第五章、結果與討論 31 第一節、探討酵母菌轉換苦瓜萃取物中皂苷之能力 31 一、菌株篩選 31 二、YCL-L-019及YCL-L-020轉換苦瓜甲醇萃取物之探討 33 第二節、YCL-L-020及YCL-L-020菌株基本特性之探討 37 一、生長溫度及世代時間 37 二、滲透壓耐受性 39 三、剛果紅耐受性 40 四、酵母菌裂解酶 (zymolyase) 敏感性 42 五、外泌蛋白組成 44 第三節、YCL-L-020中可能作用酵素之篩選 46 第四節、作用酵素之探討 49 一、質體骨架建構 51 二、酵素基因置入及轉換苦瓜皂苷能力探討 52 第六章、結論與展望 72 第七章、參考文獻 73 第八章、附錄 80 | - |
| dc.language.iso | zh_TW | - |
| dc.title | 以酵母菌發酵提高苦瓜甲醇萃取物中功效性成分3β, 7β, 25-trihydroxycucurbita-5, 23 (E)-dien-19-al之探討 | zh_TW |
| dc.title | Increasing 3β, 7β, 25-trihydroxycucurbita-5, 23 (E)-dien-19-al content in methanolic extract of Momordica charantia L. by yeast fermentation | en |
| dc.type | Thesis | - |
| dc.date.schoolyear | 107-2 | - |
| dc.description.degree | 碩士 | - |
| dc.contributor.oralexamcommittee | 黃青真;呂廷璋;陳勁初;廖辰中;邱群惠 | zh_TW |
| dc.contributor.oralexamcommittee | ;;;; | en |
| dc.subject.keyword | 苦瓜,3β,7β,25-trihydroxycucurbita-5,23(E)-dien-19-al,酵母菌,外泌蛋白,BGL1, | zh_TW |
| dc.subject.keyword | Momordica charantia,3β,7β,25-trihydroxycucurbita-5,23 (E)-dien-19-al,yeast,BGL1, | en |
| dc.relation.page | 100 | - |
| dc.identifier.doi | 10.6342/NTU201902141 | - |
| dc.rights.note | 未授權 | - |
| dc.date.accepted | 2019-07-30 | - |
| dc.contributor.author-college | 生物資源暨農學院 | - |
| dc.contributor.author-dept | 食品科技研究所 | - |
| dc.date.embargo-lift | 2029-12-31 | - |
| 顯示於系所單位： | 食品科技研究所 | |
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