黑豆咖啡被認為是一種具有降低咖啡因與養生效果的創新「雙豆飲」咖啡，而本研究以戊糖乳桿菌(Lactobacillus pentosus，簡稱LBP)發酵黑豆咖啡萃出液，探討不同黑豆與咖啡(混合)比值(10:0、7:3、5:5、3:7、0:10)與發酵時數對發酵上清液的生菌數(OD600)、pH值與抗氧化活性的影響。實驗結果顯示， LBP生菌數會隨發酵時數增加而增加，且在發酵開始後的最初4小時期間的生長速率最佳。然而LBP生菌數會隨咖啡比例的提升而減少，換言之，高黑豆含量有利於LBP生長。另外，在發酵開始後的最初4小時，發酵上清液pH值會隨黑豆在黑豆咖啡含量增加而下降速率越快，因此，黑豆萃出液對LBP而言是促進菌體生長的良好培養液。最後，不同黑豆與咖啡比值所得的發酵上清液的抗氧化活性會隨黑豆在黑豆咖啡含量增加而降低，其中以純咖啡(0:10)發酵上清液的抗氧化活性為最佳。此實驗結果顯示，不同比值之黑豆咖啡萃出液的抗氧化活性在儲存期間會隨儲存時間增加而略微逐漸下降，然而LBP活菌似乎有維持黑豆咖啡萃出液之抗氧化活性功能，不會造成黑豆咖啡萃出液之抗氧化活性隨發酵時間而有所變化。總結來看，儘管烘焙過咖啡具有良好的抗氧化活性，但卻不利於LBP的生長。然而，加入黑豆後能夠促進LBP的生長，增加菌數，有助於維持飲品的整體品質。綜合以上實驗結果，以5:5黑豆咖啡為最適比值，在此比值條件下，雖然在抗氧化活性略有降低，但仍能使LBP的生菌數保持在最適的選擇性範圍，因此達到一種平衡選擇。此研究初步呈現LBP發酵黑豆咖啡可作為促進健康的飲品，未來需要更進一步探討黑豆咖啡的抗氧化機制，並優化發酵條件以提高其抗氧化能力，而證實其具有促進健康的潛在應用價值。

Black soybean coffee is an innovative "dual-bean beverage" with the healthcare value of black soybean tea mixed with the roasted coffee for reducing caffeine. In this study, the extract of black soybean coffee was fermented by Lactobacillus pentosus (LBP), and the effects of the different ratios of black soybean to coffee (10:0, 7:3, 5:5, 3:7, 0:10) and fermentation time (hour; hr) on the viable cell count (OD600), pH, and antioxidant activity of the fermented supernatant were investigated. The results showed that the viable cell count (vcc) of LBP increased with fermentation time during the first 4 hr and the highest growth rate was observed at the first 4 hr. However, the vcc of LBP decreased with increasing in the ratio of coffee in black soybean coffee, that is, the higher content of black soybean in black soybean coffee was better for the LBP growth. Additionally, the pH of the fermented supernatant during the first 4 hr fermentation decreased and it decreased with increasing the ratio of black soybean in black soybean coffee. Therefore, the black soybean extract seemed to be suitable as the culture medium for LBP growth. Finally, the antioxidant activity of the fermented supernatant decreased with increasing the ratio of black soybean in black soybean coffee. The highest antioxidant activity of the fermented supernatant from pure coffee (0:10) was obtained and this result indicated that the extract from the roasted coffee initially had the good antioxidant activity and its activity was better than that of the extract from black soybean tea. The antioxidant activities for all the black soybean coffee extracts without LBP were slightly decreases with increasing storage time. For all the black soybean coffee extracts, LBP seems to be the good strain for keeping the antioxidant activity without significantly changing during fermentation time., therefore, LBP seems to maintain the antioxidant activity of all the fermented supernatants during fermentation. However, the addition of black soybean is good for improving the LBP growth and increasing its vcc and keeping all the quality. Finally, the optimal ratio of black soybean coffee was selected to be 5:5 due to the results of the LBP vvc, pH, and antioxidant activity, not only the antioxidant activity was slightly reduced, but also the vvc of LBP was kept within the appropriate range for achieving the balanced selection. The results of this study preliminarily indicated that the black soybean coffee fermented with LBP will be one good beverage for improving the human health. For this future research, the optimal antioxidant mechanism of black soybean coffee fermented with LBP will be obtained for the potential application in promoting health by conducting to optimize fermentation conditions and to enhance the antioxidant capacity of the fermented.

摘要..........i
Abstract..........ii
誌謝..........iv
目錄..........v
表目錄..........viii
圖目錄..........ix
第一章 緒論..........1
1.1前言..........1
1.2研究目的..........3
第二章 文獻探討..........4
2.1人口老化議題..........4
2.2台灣農產品議題..........6
2.3黑豆（Glycine max (L.) Merr.）..........7
2.3.1黑豆簡介..........7
2.3.2黑豆的營養成分及益處..........8
2.3.3黑豆的應用..........13
2.4咖啡（Coffea spp.）..........14
2.4.1咖啡簡介..........14
2.4.2咖啡的營養成分及益處..........15
2.4.3咖啡的種類..........18
2.5乳酸菌（Lactobacillus）..........19
2.5.1乳酸菌簡介..........19
2.5.2乳酸菌代謝途徑..........20
2.5.3乳酸菌的益處..........22
2.5.4乳酸菌的應用..........23
2.6戊糖乳酸菌（Lactobacillus pentosus）..........24
2.6.1戊糖乳酸菌的培養條件..........24
2.6.2戊糖乳酸菌代謝特徵..........24
2.6.3戊糖乳酸菌功能特性..........24
2.7自由基與抗氧化能力之介紹..........25
2.7.1自由基..........25
2.7.2抗氧化劑..........26
第三章 材料與方法..........27
3.1實驗材料..........27
3.2實驗藥品..........27
3.3實驗儀器..........28
3.4實驗方法..........29
3.4.1微生物培養條件..........29
3.4.2咖啡沖泡準備..........29
3.4.3發酵條件和設計..........29
3.5實驗結果分析..........31
3.5.1 pH測定..........31
3.5.2 600nm分光掃描..........31
3.5.3總還原能力測定..........32
3.5.4總酚類化合物含量..........33
3.6統計分析..........34
第四章 結果與討論..........35
4.1 pH值變化..........35
4.2 生長曲線變化（600nm）..........37
4.3還原能力試驗..........39
4.4總酚類化合物含量試驗..........42
第五章 結論與未來展望..........45
參考文獻..........47
附錄一..........55
附錄二..........56
附錄二..........57
附錄四..........58
附錄五..........59
附錄六..........60
附錄七..........61
附錄八..........62
附錄九..........63
附錄十..........64
Extended Abstract..........65
Abstract..........65

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