探討利用Lactobacillus plantarum發酵Momordica charantia山苦瓜對其降血糖及其他生物活性之影響

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研究生：	劉庭萱 Ting-Hsuan Liu
論文名稱：	探討利用Lactobacillus plantarum發酵Momordica charantia山苦瓜對其降血糖及其他生物活性之影響
指導教授：	徐敬衡
口試委員:
學位類別：	碩士 Master
系所名稱：	工學院 - 化學工程與材料工程學系 Department of Chemical & Materials Engineering
論文出版年：	2020
畢業學年度：	108
語文別：	中文
論文頁數：	105
中文關鍵詞：	山苦瓜、乳酸菌發酵、降低血糖、抑制澱粉酶
外文關鍵詞：	α-amylase inhibition
相關次數：	點閱：10 下載：0
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山苦瓜（Momordica charantia）因具有保健及藥理學功效，近年來被廣泛地進行研究，其中有多篇文獻指出山苦瓜能有效地降低血糖，可被應用於治療第二型糖尿病。然而雖有許多專注於山苦瓜生物活性功能之研究，對於山苦瓜透過微生物發酵的文獻卻相對較少。微生物的發酵製程不僅可以改善風味、延長食品的保存期限，亦可改變原料的生物活性，進而提升其營養及經濟價值。因此本研究利用胚芽乳酸菌Lactobacillus plantarum對山苦瓜進行發酵，欲了解山苦瓜成分與生物活性之關係，並提升其降低血糖及抗氧化之能力。
好氧厭氧、培養溫度及營養源皆是影響微生物生長及表現的因素，對發酵山苦瓜的成份及生物活性產生影響，因此本研究將探討不同培養方式、溫度及以豆漿作為營養源之不同濃度下，利用Lactobacillus plantarum BCRC15478發酵山苦瓜對其成份及生物活性之影響。
苦瓜素（Charantin）為山苦瓜中的一種類固醇糖苷化合物，能對碳水化合物代謝酶進行抑制作用，故本研究觀察此成分在發酵過程中的變化，並與作為抗糖尿病指標的α-澱粉酶抑制率相比較。結果顯示經發酵，微生物分解苦瓜細胞壁，釋放出更多苦瓜素，進而對山苦瓜提取液貢獻更好的澱粉酶抑制效果。此外，亦發現山苦瓜發酵液及提取液中的抗氧化性與多酚物質相關，發酵後可有效提升抗氧化活性。經由實驗結果，L. plantarum於30°C厭氧培養下添加10 %豆漿作為營養源，有最好的菌種生長狀況，且發酵後的山苦瓜液具91.55 %的DPPH自由基清除力，山苦瓜乾粉提取液則有47.14 %的DPPH自由基清除力及64.75 %的α-澱粉酶抑制活性，相比未發酵前增加37.07 %的苦瓜素含量。

Various biological activities and pharmacological effects of Momordica charantia have been reported recent years. Lots of them demonstrated that bitter melon can used for lowering blood glucose levels in patients with diabetes mellitus. However, although there are many studies focusing on the biological activity of M. charantia, there is no sufficient literature to use on the microbial fermentation. Fermentation process not only improved the flavor of the raw material, but also extended the shelf life of the food and enhanced its nutritional value and economic value. Therefore, M. charantia fruit was used to fermented by Lactobacillus plantarum to enhanced its anti-diabetes and antioxidant activities. Moreover, the relationship between compounds and bioactive functions will be discuss in this study.
Charantin is a steroidal glycoside present in the Momordica charantia which can inhibit the key enzymes related to carbohydrate metabolism. Thus, the present study aimed to investigate charantin content and α-amylase inhibition ability during the fermentation. Research is also focusing on total phenolic compounds and antioxidant ability via fermentation with L. plantarum. Results showed that inhibition of α-amylase activity and DPPH scavenging activity can be improved in fermented M. charantia juice. The M. charantia juice with 10 % soymilk as a nutrient supplementation under 30°C provide good environment for the growth of cell viability. The fermented juice and the extraction of fermented M. charantia powder showed higher DPPH scavenging activity and α-amylase inhibition activity compared with the unfermented juice.

摘要    i
Abstract    ii
誌謝    iii
目錄    iv
圖目錄    viii
表目錄    xi
一、緒論    1
1-1 研究動機    1
1-2 研究目的    2
二、文獻回顧    3
2-1山苦瓜    3
2-1-1山苦瓜的基本介紹    3
2-1-2山苦瓜的成份    3
2-1-3山苦瓜的生物活性及功效    6
2-2 苦瓜素（Charantin）    9
2-2-1 苦瓜素的基本介紹    9
2-2-2 苦瓜素的功效    11
2-2-3 苦瓜素的萃取方法    12
2-3 糖尿病（Diabetes）    12
2-3-1 糖尿病的基本介紹    12
2-3-2 糖尿病的成因和類別    13
2-3-3 第2型糖尿病的治療方法    13
2-4 乳酸菌發酵    15
2-4-1 乳酸菌發酵的基本介紹    15
2-4-2 乳酸菌發酵進行生物轉化    15
2-5 山苦瓜發酵研究    18
三、材料與方法    21
3-1 實驗規劃    21
3-2 實驗材料    23
3-2-1 實驗菌株    23
3-2-2 實驗藥品    24
3-2-3 實驗儀器與設備    26
3-3 實驗方法    28
3-3-1菌種保存及固態培養    28
3-3-2液態種瓶培養    29
3-3-3液態發酵實驗    30
3-3-4苦瓜素萃取條件實驗    32
3-4 分析方法    34
3-4-1 平板菌落計數法    34
3-4-2 pH值分析    34
3-4-3 還原糖濃度分析    35
3-4-4 乳酸分析    37
3-4-5 總多酚類化合物含量測定（Total phenolic compounds）    39
3-4-6 蛋白質濃度測定（BCA protein assay）    40
3-4-7 DPPH自由基清除能力測定（DPPH scavenging activity）    42
3-4-8 α-澱粉酶抑制能力測定（α-amylase inhibition ability）    42
3-4-9 苦瓜素濃度分析    44
四、結果與討論    46
4-1 Charantin苦瓜素萃取條件最適化    46
4-2 Lactobacillus plantarum的生長曲線    49
4-3 不同發酵方式對山苦瓜之影響    50
4-3-1 發酵方式對Lactobacillus plantarum生長的影響    50
4-3-2 發酵方式對Momordica charantia發酵液生物活性的影響    53
4-3-3 發酵方式對Momordica charantia萃取液生物活性的影響    55
4-3-4發酵方式對發酵山苦瓜之結論    56
4-4 發酵溫度對山苦瓜之影響    57
4-4-1發酵溫度對pH值和殘糖濃度的影響    57
4-4-2發酵溫度對發酵液抗氧化活性及總多酚含量的影響    59
4-4-3發酵溫度對萃取液抗氧化活性及總多酚含量的影響    62
4-4-4發酵溫度對發酵液澱粉酶抑制活性及蛋白質含量的影響    64
4-4-5發酵溫度對萃取液澱粉酶抑制活性及苦瓜素含量的影響    66
4-4-6發酵溫度對山苦瓜發酵液及萃取液生物活性影響之結論    68
4-5 添加稀釋豆漿對發酵山苦瓜之影響    72
4-5-1稀釋豆漿濃度對發酵山苦瓜的影響    72
4-5-2 添加稀釋豆漿對Lactobacillus plantarum生長狀態的影響    74
4-5-3 添加稀釋豆漿對pH值、殘糖濃度和乳酸含量的影響    75
4-5-4添加稀釋豆漿對發酵液抗氧化活性的影響    77
4-5-5添加稀釋豆漿對萃取液抗氧化活性的影響    79
4-5-6添加稀釋豆漿對發酵液抑制澱粉酶活性的影響    80
4-5-7添加稀釋豆漿對萃取液抑制澱粉酶活性的影響    81
4-5-8添加稀釋豆漿對萃取液中苦瓜素含量的影響    82
4-5-9添加稀釋豆漿對山苦瓜發酵液及萃取液生物活性比較    83
五、結論    85
參考文獻    87
                                

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