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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 羅翊禎 | |
| dc.contributor.author | Yun-Ju Lai | en |
| dc.contributor.author | 賴韻如 | zh_TW |
| dc.date.accessioned | 2021-06-17T03:12:07Z | - |
| dc.date.available | 2023-07-19 | |
| dc.date.copyright | 2018-07-19 | |
| dc.date.issued | 2018 | |
| dc.date.submitted | 2018-07-16 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/69286 | - |
| dc.description.abstract | 苦瓜(Momordica charantia)為葫蘆科一年生草本植物，文獻指出苦瓜皂苷是苦瓜苦味及部分功效成分來源，為一群葫蘆烷型三萜類。本研究目的希望透過酵母菌發酵或乳酸菌之醣苷水解酵素來減少苦瓜中苦味皂苷含量。由於苦瓜皂苷骨架由具長鏈側枝的四環三萜構成，多為不帶醣基之皂苷元或修飾有一至兩個醣基之醣苷態。此類結構之皂苷於培養基質中之溶解度不佳，本實驗首先使用大孔吸附樹脂Diaion™ HP-20針對甲醇萃取物進行純化，收集70-100%梯度甲醇可流洗出之苦瓜皂苷，再透過Tween 20增加其溶解度，經微生物生長曲線及點試驗後，選定以含有0.05% Tween 20之苦瓜皂苷培養基進行後續轉換。經Saccharomyces cerevisiae BY4741和Lactobacillus plantarum BCRC 10069轉換後，觀察到兩者之苦瓜皂苷於轉換前後皆有波鋒消長情形，進一步分析後發現經兩者發酵後苦味皂苷Momordicoside K和Momordicine I皆有顯著下降趨勢。為確認是否為醣苷水解酶作用，於酵母菌選定缺乏合成醣苷水解酶基因之菌株進行發酵，而乳酸菌則透過表現其putative β-glucosidase LpBgl進行試驗，分析後皆發現苦瓜皂苷之轉換與鎖定的目標酵素皆無直接相關。最後為得知苦瓜皂苷之轉換情形，使用菌株發酵單一苦瓜皂苷進行觀察，發現在酵母菌與乳酸菌皆發現Momordicoside K含量之減少除轉換成皂苷元外，尚有一部份會轉換成Momordicoside L，可能為甲基轉移酶(transmethylases)或去甲基酶(demethylases)作用所致。綜合上述結果，可以發現酵母菌及乳酸菌中皆有可轉換苦瓜苦味皂苷之酵素，後續可再針對進行轉換之酵素進行探討及應用。 | zh_TW |
| dc.description.abstract | Momordica charantia is a member of the Curcurbitaceae family and is commonly known as bitter melon. Previous studies indicated that specific bitter melon saponins were contributed to the bitter taste and functional activities. The purpose of this study is to reduce the contents of bitter taste saponins by using yeast fermentation or glucosidase hydrolysis. Enhancing the solubility of bitter melon saponins by adding 0.05% Tween 20 is necessary due to the low polarity of tetracyclic triterpenoids of the bitter melon saponins. The results showed that the composition of saponins decline after fermentation with Saccharomyces cerevisiae or with Lactobacillus plantarum. Specifically, the contents of bitter taste saponins, momordiciside K and momordicine I decreased significantly after fermentation. To dissect the underlying mechanism, we used glucosidase deleted yeast to undergo fermentation and in vitro study with putative glucosidase LpBgl in LAB. Unfortunately, we did not identify specific glucosidase related to the hydrolysis of saponins. However, we discovered that momordicoside K was converted into aglycone and the other became momordicoside L. This results indicated that not only glucosidase involved in the biotransformation of saponins, but also possibly transmethylases or demethylases. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-17T03:12:07Z (GMT). No. of bitstreams: 1 ntu-107-R05641004-1.pdf: 5096176 bytes, checksum: b5156c39c2f7d4d54d5598888c29c236 (MD5) Previous issue date: 2018 | en |
| dc.description.tableofcontents | 壹、前言 1
貳、文獻回顧 2 第一節、 苦瓜 2 第二節、 苦瓜皂苷 2 一、化學結構 3 二、生合成途徑 5 三、生理活性 5 四、苦味皂苷 7 第三節、 生物轉換 9 一、葡萄醣苷酶(β-glucosidase) 9 二、酵母菌中的葡萄醣苷酶(β-glucosidase) 9 三、乳酸菌中的葡萄醣苷酶(β-glucosidase) 11 第四節、 苦瓜皂苷的提取 12 一、大孔吸附樹脂(Macroporous adsorption resins) 12 第五節、 苦瓜皂苷的分析方法 14 一、比色法 14 二、薄層層析法 14 三、高效能液相層析法(HPLC) 15 參、研究目的與實驗架構 17 肆、材料與方法 20 第一節、實驗材料 20 一、山苦瓜 20 二、酵母菌菌株 20 三、乳酸菌菌株 20 四、質體 21 三、微生物培養材料 22 第四節、實驗方法 25 一、苦瓜皂苷生物轉換及分析平台之建立 25 二、透過生物轉換苦瓜皂苷萃取物 29 伍、結果與討論 33 第一節、苦瓜皂苷生物轉換及分析平台之建立 33 一、以六種苦瓜皂苷標準品確立苦瓜皂苷分子離子類型 33 二、苦瓜皂苷的檢量線線性、方法檢量之偵測極限與定量極限 35 三、使用Diaion™ HP-20純化苦瓜皂苷甲醇萃取物 35 四、透過Tween 20增加苦瓜皂苷於培養基質中之溶解度 38 五、以固相萃取管純化苦瓜皂苷培養基質之回收率 43 第二節、以酵母菌Saccharomyces cerevisiae BY4741轉換苦瓜皂苷萃取物之探討 45 一、使用野生型酵母BY4741 WT進行苦瓜皂苷萃取物之發酵 45 二、使用單一基因剔除菌株進行苦瓜皂苷萃取物之發酵 51 三、使用Saccharomyces cerevisiae BY4741 WT發酵單一苦瓜皂苷 60 第三節、以乳酸菌Lactobacillus plantarum BCRC10069轉換苦瓜皂苷萃取物之探討 66 一、使用常見之植物性乳酸菌L. plantarum BCRC10069進行苦瓜皂苷萃取物汁發酵 66 二、乳酸菌β-葡萄醣苷酶(Bgl)蛋白表現及活性 72 三、以乳酸菌蛋白LpBgl進行苦瓜皂苷萃取液胞外發酵 79 四、使用Lactobacillus plantarum BCRC 10069發酵單一苦瓜皂苷 81 陸、結論與展望 87 柒、參考文獻 89 捌、附錄 104 | |
| dc.language.iso | zh-TW | |
| dc.subject | 生物轉換 | zh_TW |
| dc.subject | Lactobacillus plantarum | zh_TW |
| dc.subject | Saccharomyces cerevisiae | zh_TW |
| dc.subject | 苦味皂? | zh_TW |
| dc.subject | 苦瓜 | zh_TW |
| dc.subject | biotransformation | en |
| dc.subject | cucurbitane-type saponins | en |
| dc.subject | bitter taste | en |
| dc.subject | Saccharomyces cerevisiae | en |
| dc.subject | Lactobacillus plantarum | en |
| dc.subject | Momordica charantia | en |
| dc.title | 探討酵母菌與乳酸菌轉換苦瓜皂苷之影響 | zh_TW |
| dc.title | The effect of Saccharomyces cerevisiae and Lactobacillus plantarum BCRC 10069 biotransformation on saponins of Momordica charantai L. | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 106-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 呂廷璋,陳勁初,廖家慶,黃學聰,王如邦 | |
| dc.subject.keyword | 苦瓜,苦味皂?,Saccharomyces cerevisiae,Lactobacillus plantarum,生物轉換, | zh_TW |
| dc.subject.keyword | Momordica charantia,cucurbitane-type saponins,bitter taste,Saccharomyces cerevisiae,Lactobacillus plantarum,biotransformation, | en |
| dc.relation.page | 138 | |
| dc.identifier.doi | 10.6342/NTU201801347 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2018-07-16 | |
| dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
| dc.contributor.author-dept | 食品科技研究所 | zh_TW |
| 顯示於系所單位： | 食品科技研究所 | |
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