臺灣博碩士論文加值系統

自民國八十九年四月十九日頒布菸酒管理法以來，已有百餘家民間酒莊成立。篩選台灣本土釀酒酵母菌，發展高品質且具特殊風味酒產品有其必要。分別自梅子、荔枝、葡萄、黑后葡萄、柳丁、芒果、鳳梨、楊桃及龍眼等水果自然發酵酒醪中共篩得364株酵母菌，其中123株具有較高的CO2產量(>10 mL/day)，測定其酒精產率後，以篩選自黑后葡萄HP01、HP04、HP06、HP10、HP15、HP20及HS13，及以龍眼為分離源所得之DM02、DM08及DH09等10株酵母菌株，酒精產率高於5.0 %(v/v)。10株酵母菌株接種於含24% (w/w)蔗糖的YMP broth中25 ℃培養10天後，酒精含量高於5.0 %(v/v)，發酵液濁度低於20 NTU，揮發性酸含量低於1.2 g/L，應可用於黑后葡萄及龍眼水果酒的釀製。
黑后葡萄為台灣釀製紅葡萄酒主要原料之一，篩選自黑后葡萄酒醪的7株酵母菌具有較高之酒精產量，經Biolog system鑑定均為Saccharomyces cerevisiae。上述酵母菌株分別接種於黑后葡萄泥中於25 ℃發酵14天後，酒精產量可達15 % (v/v)，揮發性酸產量低於0.7 g/L。與三株商業酵母菌A、B及C相比，以HP01為菌酛發酵之黑后葡萄酒具有最高的酯類含量、顏色強度、嚐味及整體接受度，而以HS13為菌酛者則具有最高酒精、高級醇、揮發性脂肪酸及酚類含量。HP01及HS13之粒腺體DNA經限制酶水解後，其片段組成與多株酵母菌株有明顯差異。HP01及HS13之優良發酵特性，應可用於黑后葡萄酒之釀製，發展具特殊風味之紅葡萄酒。
以米糠、麩皮、蜂蜜、脫脂乳及sodium glutamate等保護劑進行冷凍乾燥黑后葡萄酒菌酛的製備，結果發現HP01添加麩皮經凍乾後，其存活率可較未添加保護劑者高2 倍，HP01添加10 %(w/w) 麩皮及20 %(w/w)脫脂乳，HS13添加20 %(w/w)脫脂乳、5 %(w/w) 蜂蜜及5%(w/w) sodium glutamate之處理組，分別可達到最高的存活率51及80 %。凍乾酵母菌酛與濕酵母菌酛發酵時間無明顯差異，且製成紅葡萄酒之品質特性亦無明顯差異。而於菌酛保存試實驗中，凍乾酵母菌酛經真空包裝後，貯存於4 ℃下，可保存15 個月，故可供應酒庄一可長期保存且方便使用的葡萄酒菌酛。

More than hundreds of private wineries have been established since the Taiwan Tobacco and Alcohol Administration Act was enacted on April 19, 2000. It is essential to screen local wine yeast in order to develop quality wine with unique flavor. 364 yeasts were first isolated from plum, lichee, grape, Black Queen grape, orange, mango, pineapple, carambola and longan must with the supplement of sugar to 17 and 30。Brix, respectively. Among these yeasts, 123 yeasts with CO2 productivity greater than 10 mL after 24 h incubation in YMPG broth containing 10 % glucose were considered as potential yeast isolates. These 123 yeasts were then evaluated for their ethanol productivity in YMPS broth containing 20 % sucrose. Ten yeasts form Black Queen grape and longan with ethanol productivity greater than 5.0 % (v/v) selected for further fermentation properties test. Ethanol contents of fermented YMPS broth were all above 5.0 %(v/v), volatile acidity and turbidity all below 1.2 g/L and 20 NTU, may useable as Black Queen grape and longan wine starter.
Black Queen grape is a main ingredient for red wine brewing in Taiwan. Seven yeasts identified as Saccharomyces cerevisiae by Biolog System with higher ethanol productivity were individually inoculated into Black Queen grape must to undergo fermentation at 25℃ for 14 days. The ethanol contents and volatile acidity in wine were above 15% (v/v) and below 0.7 g/L, respectively. Compared to commercial wine yeasts A, B and C, HP01 fermented red wine contained the highest total ester content, color intensity, taste grade, and overall preference value. On the other hand, HS13 fermented red wine contained the highest ethanol, higher alcohols, fatty acids and phenolic compounds content. The mtDNA restriction fragments profiles of these two wine yeast strains were different from those of other yeast strains. Due to their superiority in oenological characteristics, HP01 and HS13 can be applied in Black Queen red wine brewing to produce red wine with unique flavor.
The effects of several protectants such as rice hull, wheat bran, skim milk, trehalose, and sodium glutamate on the viability of freeze-dried Saccharomyces cerevisiae strains HP01 and HS13 and their fermentation properties such as pH, volatile acidity, ethanol content, titratable acidity, color intensity, phenolic content and residual sugar during grape wine production were investigated. Besides, the effects of storage at 4℃ and 25℃ for 1 to 15 months on the viability and alcoholic fermentation properties of freeze-dried cells and wet cells were also studied. Results showed that the supplement of 10% wheat bran in freeze-drying menstrua could significantly increase the survival rate of freeze-dried HP01 from 25% to 46%. The simultaneous supplement of 10% wheat bran and 20% skim milk increased the survival rate of freeze-dried HP01 to 51%. For the freeze-drying of HS13, the simultaneous supplement of 20% skim milk, 5% honey, and 5% sodium glutamate showed the greatest survival rate, 80% while the survival rate of its control was 9.2%. The fermentation properties of freeze-dried yeasts were like their freshly prepared wet cells; namely, both of them produced 15% ethanol after 9 days fermentation at 25℃. At 4℃ storage, freeze-dried HP01 and HS13 could maintain their viability and fermentation properties as their wet cells for 15 months. While storing at 25℃, the storage lives of freeze-dried cells were less than 2 months.

口試委員會審定書 i
誌謝 ii
中文摘要 iv
英文摘要 v
第一章 文獻整理 1
壹、釀酒技術簡介 1
貳、釀酒用酵母菌簡介 8
參、酒中風味物相關物質 13
肆、酵母菌的發酵生理及風味物質的生合成 24
伍、釀酒用酵母菌的篩選 34
陸、釀酒用酵母菌菌種改良 38
柒、酒精飲料與人體健康 41
捌、參考文獻 46
第二章 水果酒釀造用酵母菌收集及篩選 50
壹、前言 50
貳、材料與方法 51
酵母菌分離保存 51
酵母菌產氣及發酵試驗 51
酵母菌發酵特性分析 51
參、結果與討論 56
酵母菌株的收集及產氣試驗 56
酵母菌株酒精產率分析 56
酵母菌發酵特性分析 67
結論 69
肆、參考文獻 69
第三章 黑后葡萄酒釀酒用酵母菌株鑑定及發酵特性分析 72
壹、前言 72
貳、材料與方法 73
酵母菌株篩選與保存 73
少量釀造試驗 73
酵母菌釀造特性分析 74
黑后葡萄酒嗜好性品評 76
黑后葡萄酒香氣成分萃取、鑑別及定量 76
酵母菌株粒腺體DNA限制酶分析 77
統計分析 78
參、結果與討論 78
酵母菌株鑑定 78
以HP01及HS13與商業釀酒用酵母為菌酛釀製之黑后葡萄酒
品質特性比較 83
以HP01及HS13與商業釀酒用酵母為菌酛釀製之黑后葡萄酒
揮發性成分分析 85
HP01及HS13之粒腺體DNA限制酶分析 89
結論 90
肆、參考文獻 93
第四章凍乾黑后葡萄酒釀酒用菌酛之製備 97
壹、前言 97
貳、材料與方法 99
使用菌種及試藥 99
冷凍乾燥活性乾酵母及濕酵母製備 99
酵母菌計數 100
酵母菌酛乾重及水分測定 100
黑后葡萄酒少量發酵試驗 101
酵母菌株發酵特性分析 101
酵母菌酛保存性試驗 102
統計分析 103
參、結果與討論 103
凍乾酵母菌酛存活率 103
凍乾酵母菌酛發酵特性分析 107
凍乾酵母菌酛保存試驗 110
結論 112
肆、參考文獻 115
附錄Selection and Oenological Comparison of Taiwan
Black Queen Grape Wine Yeasts (已接受) 116

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