壹、前言 1
貳、文獻整理 3
一、咖啡 3
1. 咖啡簡介 3
2. 咖啡加工 (Coffee processing) 4
3. 副產物 (By-products) 4
4. 咖啡風味 5
二、咖啡渣 (Spent coffee grounds, SCG) 6
1. 成分組成 6
1.1 碳水化合物 6
1.2 蛋白質 7
1.3 脂質 7
1.4 礦物質 8
1.5 酚類化合物 8
1.6 非蛋白質含氮化合物 9
2. 咖啡渣成分與降血糖活性 10
3. 咖啡渣發酵 11
三、糖尿病 13
1. 糖尿病簡介 13
2. 糖尿病診斷 13
3. 肝糖合成與糖質新生 13
4. 胰島素的訊息傳遞與分子 14
參、實驗架構 15
肆、實驗材料與方法 16
一、材料 16
1. 原料 16
2. 水解酵素 16
3. 菌株與細胞株 16
3.1 實驗菌株 16
3.2 細胞株 17
4. 藥品 17
5. 培養基 18
5.1 菌株培養基 18
5.2 細胞培養基 18
二、儀器設備 18
三、實驗方法 19
1. 咖啡渣酵素前處理 19
1.1 醣解酵素篩選 19
1.2 醣解酵素水解溫度 20
1.3 醣解酵素濃度 20
1.4 醣解酵素水解時間 20
2. 菌株保存與活化 20
2.1 菌株保存 20
2.2 菌株活化 21
3. 咖啡渣發酵條件探討 21
3.1 乳酸桿菌及酵母菌株之篩選 21
3.2 菌株相容性試驗 21
4. 分析 22
4.1 一般成分分析 22
4.1.1 水分 22
4.1.2 灰分 22
4.1.3 粗脂肪 22
4.1.4 粗蛋白 22
4.1.5 粗纖維 23
4.1.6 碳水化合物 23
4.2 菌數分析 23
4.3 pH值 24
4.4 可滴定酸度分析 24
4.5 總胜肽含量 24
4.6 還原糖含量分析 25
4.7 總酚含量分析 25
4.8 咖啡因含量分析 25
4.9 綠原酸含量分析 25
4.10 抑制α-Glucosidase活性分析 26
4.11 抑制α-Amylase活性分析 26
4.12 色差分析 27
4.13 感官品評分析 28
5. 以FL83B細胞模式分析發酵產物之降血糖活性 29
5.1 FL83B細胞培養 29
5.2 不同濃度樣品對FL83B細胞存活率影響 (MTT assay) 29
5.3 誘導 FL83B 細胞產生胰島素阻抗並測定葡萄糖利用率 30
6. 統計分析 30
伍、結果討論 31
一、咖啡粉與咖啡渣之一般組成 31
二、酵素水解咖啡渣與水解條件探討 31
三、微生物發酵咖啡渣水解液之篩選與評估其降血糖潛力 33
四、乳酸菌和酵母菌共同發酵與其相容性試驗 35
1. 乳酸菌株與酵母菌株PK之複合菌株發酵 35
2. 乳酸菌株與酵母菌株SP54之複合菌株發酵 37
五、發酵咖啡渣萃取物之降血糖活性與生理活性物質分析 39
1. 咖啡渣萃取物對FL83B細胞存活率分析 39
2. 發酵咖啡渣萃取物對具胰島素阻抗FL83B細胞葡萄糖利用率之影響 39
3. 發酵咖啡渣萃取物之生理活性物質含量分析 41
六、發酵咖啡渣萃取液之色差分析與感官品評 42
陸、結論 45
柒、參考文獻 46

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