臺灣博碩士論文加值系統

摘要
本研究以滅菌的洋蔥(Allium cepa L.)汁為材料，依序進行酒精
和醋酸的批式發酵試驗，包括篩選出二株最適合洋蔥酒發酵的酵母菌與醋酸發酵的醋酸菌及建立洋蔥醋的發酵模式與相關技術的探討。
洋蔥含有Allysine的成分具有辛辣而刺激性的味道，且有抑菌作用。在預實驗時，酵母菌幾乎無法在生洋蔥汁中行酒精發酵。然而，Allysine的成分具有揮發性，可在殺菌釜滅菌過程去除，故以滅菌
的洋蔥汁即可成功進行酒精發酵。
酵母菌株之篩選以Saccharomyces cerevisiae CCRC 21551株之最終酒精濃度最高，達14.1%(v/v)，而Saccharomyces cerevisiae CCRC 20262具有特殊的水果香味，故以這兩株酵母菌進行最適酒精發酵條件之探討。
醋酸菌的篩選以Acetobactor sp. CCRC 12326產醋效率最高，可在含5%酒精的Mannitol medium中發酵五天產出4.86%(v/v)的醋酸，故以之進行最適醋酸發酵條件之探討。選用洋蔥品種CAL 505的洋蔥汁為原料，其可溶性固形物為8.4∘Brix，可發酵成酒精濃度3.92%(v/v)的洋蔥酒，再發酵為醋酸濃度3.43%(v/v)的洋蔥醋。
在不同濃度洋蔥酒及添加醋酸與否的醋酸發酵比較試驗中，發酵第六天以5%洋蔥酒產醋酸較高，為3.44%(v/v)，而發酵第八天以7%洋蔥酒產醋酸較高，為3.61%(v/v)；添加1%醋酸的洋蔥酒其醋酸發酵效率並未增加。
在以添酒方式行醋酸發酵之試驗中，經二次添酒後到第十五天其滴定醋酸達3.4829%，而其體積因二次添酒而增加為2.25倍。而以二階段馴化方式行醋酸發酵，可使產醋效率提高及酸度增加，其中以菌酛及種醋培養各三天的方式，經24天發酵之實際產醋量最高為4.97%；而菌酛及種醋培養各五天者，最終醋酸濃度可達6.02%(v/v)。
洋蔥醋的其它有機酸成分為每一百毫升含葡萄糖醛酸0.43克、葡萄糖酸0.91克、酒石酸0.16克、焦麩胺酸0.097克及蘋果酸0.80克。

Abstract
The objectives of this study were to investigate the alcohol and acetic acid fermentations using the sterile onion(Allium cepa L.) juice as raw material and performed by traditional static cultivation system. This included screening of yeast strain and acetic acid bacteria strain, and establishment of the conditions for onion vinegar fermentation process.
Allysine compounds present in onion have a keen, stimulating smell and an anti-bacterial property, which inhibits the growth of yeast. In preliminary experiments, difficulty was experienced in fermenting the raw onion juice using yeast. However, since the allysine compounds are volatile and they can be removed by autoclaving. Alcohol and acetic acid
fermentations using the sterile onion juice were thus performed successfully by batch culture.
Saccharomyces cerevisiae CCRC 21551 was chosen for its best alcohol producing capability, up to 14.1%(v/v) was obtained, while S. cerevisiae 20262 was best for its special fruit flavor . These two strains were chosen to study the subsequent determination of optimum condition of alcohol fermentation.
Acetobacter sp. CCRC 12326 was chosen to study the optimum condition of acetic acid fermentation. The acetic acid producing capability of this strain was up to 4.86% (v/v) in mannitol medium containing 5% ethanol.
The juice from the onion CAL 505 cultivar, which contain 8.4 ºBrix soluble solid, was converted by yeast to onion wine containing 3.92% (v/v) ethanol; and then fermented further to onion vinegar with 3.43% (v/v) acetic acid.
Evaluations of the fermentation efficiencies for Acetobacter sp. CCRC 12326 on fermenting onion wines (OWs) with different concentrations of ethanol, and OWs with or without supplementing with 1% acetic acid was performed. The highest acidity of onion acidity vinegar in the 6th day was 3.44% (v/v) from the OW containing 5% ethanol, and the highest one in the 8th day was 3.61% from the OW with 7% alcohol. However, supplementing the OW with 1% acetic acid did not enhance the vinegar fermentation rate.
The acidity of onion vinegar was 3.48% at day 15 if the fresh OW was added during fermentation, and accompanied by the increase of final volume of broth to 2.25 times.
Both acetic acid production rate and acidity increased if a modified two-stage vinegar fermentation method which comprised four operation modes(A1, A2, B1, B2) was used. The highest acetic acid production rate was obtained for the vinegar fermentation performed according to A1 mode (starter-making 3 days, mother vinegar-making 3 days); the concentration of acetic acid was 4.97% after 24 days. However, the final acidity was higher, 6.02%, for that of using B2 mode (starter-making 5 days and mother vinegar-making 5 days).
The contents of major organic acids except acetic acid in the onion vinegar were glucuronic acid 0.43, gluconic acid 0.91, tartaric acid 0.16, pyroglutamic acid 0.097, and malic acid 0.80 g/100ml.

目錄
壹、前言................................................................................................ 1
貳、文獻整理........................................................................................ 3
【壹】洋蔥的相關介紹………………………………………............. 3
一、洋蔥的組成分…………………………………………............. 3
二、洋蔥的抑菌性…………………………………………............. 3
三、洋蔥的保健功效………………………………………............. 5
(一)洋蔥中含硫成分的保健功效………………………............. 5
(二)洋蔥中類黃酮的保健功效…………………………............. 9
四、洋蔥的加工製品………………………………….................. 13
【貳】醋酸發酵之相關介紹…………………………………........... 16
一、醋的定義及功能……………………………………................. 16
(一)醋的定義……………………………………………….......... 16
(二)醋的功能…………………………………………….............. 16
二、醋酸菌之研究……………………………………………........ 17
(一)醋酸菌的分離與培養…………………………...................... 18
(二)醋酸菌的種類……………………………………….............. 18
三、食醋之釀造方法………………………………………............ 19
(一)傳統的靜置發酵…………………………………….............. 19
(二)深部培養法(Submerged culture) …………………................ 20
四、影響醋酸發酵的因素…………………………………............ 22
(一)氧氣………………………………………………….............. 23
(二)溫度………………………………………………….............. 23
(三)發酵基質成份比例…………………………………….......... 23
(四)醋酸發酵的促進因子……………………………………...... 25
參、材料與方法………………………………………………........... 26
【壹】實驗材料…………………………………………………...... 26
一、洋蔥………………………………………………………....... 26
二、實驗菌株………………………………………………........... 26
三、培養基………………………………………………….......... 26
四、化學藥劑……………………………………………….......... 27
五、儀器設備……………………………………………….......... 28
六、電腦套裝軟體........................................................................... 29
【貳】實驗方法……………………………………………........... 30
一、實驗大綱…………………………………………….…...... 30
二、實驗方法………………………………………………....... 30
(一) 洋蔥汁的製備………………………………………....... 30
(二)酒精發酵條件之探求…………………………………..... 31
(三)醋酸發酵條件之探求..…………………………………...... 32
三、分析方法………………………………………………...... 33
四、統計分析及圖形繪製…………………………………...... 35
肆、結果與討論………………………………………………....... 36
【壹】最適酵母菌株之篩選……………………………………..... 36
【貳】最適醋酸菌菌株之篩選…………………………………..... 49
【參】醋酸發酵條件之探討……………………………………..... 56
(一)不同酒精濃度洋蔥酒及添加醋酸與否之醋酸發酵比較….. 56
(二)以12 o與14oBrix SSOJ發酵之洋蔥酒行醋酸發酵比較…… 59
(三)以添酒方式行醋酸發酵試驗。…………………………….... 62
(四)二階段馴化方式醋酸發酵比較試驗。…………………….... 64
【肆】其他成份分析結果………………………………………..... 73
(一)糖類分析………………………………………………........ 73
(二)其他有機酸分析(除醋酸外) ……………............................. 79
伍、結論…………………………………………………………....... 82
參考文獻…………………………………………………………..... 84
圖目錄
圖次
圖一、不同酵母菌菌株於滅菌洋蔥汁中培養生菌數之變化.... 37
圖二、不同酵母菌菌株於滅菌洋蔥汁中培養生菌數之變化.... 38
圖三、不同酵母菌菌株於滅菌洋蔥汁中培養生菌數之變化.... 39
圖四、不同酵母菌菌株於滅菌洋蔥汁中培養生菌數之變化.... 40
圖五、不同酵母菌菌株於補糖洋蔥汁(SOJ)靜置發酵時第三
和第五天之產酒精變化..................................................
43
圖六、六株優勢酵母菌株於補糖洋蔥汁(SSOJ)中靜置發酵
時之產酒精變化..............................................................
44
圖七、三株優勢酵母菌株於補糖洋蔥汁(SSOJ)中靜置發酵
時之生菌數變化..............................................................
45
圖八、三株優勢酵母菌株於補糖洋蔥汁(SSOJ)中靜置發酵
時之生菌數變化..............................................................
46
圖九、兩株選定酵母菌個別或混合於滅菌洋蔥汁靜置發酵
之產酒精變化..................................................................
47
圖十、兩株選定酵母菌個別或混合於滅菌洋蔥汁靜置發酵
之生菌數變化..................................................................
48
圖十一、不同醋酸菌發酵過程產醋量與殘存酒精的變化........ 51
圖十二、不同醋酸菌發酵過程生菌數變化................................ 52
圖十三、不同醋酸菌發酵過程pH值變化.................................. 54
圖十四、Acetobacter sp. CCRC 12326發酵過程產醋量與殘
存酒精量的變化..........................................................
55
圖十五、添加醋酸對不同酒精濃度洋蔥酒醋酸發酵之影響... 57
圖十六、添加醋酸對不同酒精濃度洋蔥酒醋酸發酵時pH變
化之影響......................................................................
60
圖十七、以12 o與14oBrix洋蔥汁為原料所製得之洋蔥酒行醋
酸發酵時酸度與pH之變化比較.................................
61
圖十八、以添酒方式行醋酸發酵之滴定酸度變化.................... 63
圖十九、二階段馴化方式醋酸發酵之生菌數變化.................... 65
圖二十、二階段馴化方式醋酸發酵之醋酸與酒精濃度變化.... 66
圖二十一、二階段馴化方式醋酸發酵之醋酸與酒精濃度變
化..............................................................................
69
圖二十二、二階段馴化控制組之醋酸與酒精濃度比較........... 70
圖二十三、二階段馴化方式醋酸發酵之實際產醋量比較........ 71
圖二十四、兩株選定酵母菌個別及混合行酒精發酵之甘油
副產物變化..............................................................
75
圖二十五、不同糖度洋蔥汁行酒精發酵之甘油副產物變化.... 76
圖二十六、不同糖度洋蔥汁完全發酵產生之酒精與甘油的
關係.......................................................................
78
表目錄
表次
表一、不同酵母菌株於滅菌洋蔥汁中培養時殘糖之變化.......... 41
表二、不同醋酸菌於含5%酒精之Mannitol培養基中發酵第
五天之醋酸含量................................................................
53
表三、添加醋酸對不同酒精濃度洋蔥酒醋酸發酵之影響......... 58
表四、以二階段馴化方式行醋酸發酵之產醋量比較(HPLC分
析值)...................................................................................
68
表五、洋蔥製品的HPLC糖類分析.............................................. 74
表六(a)、洋蔥製品的有機酸含量比較(除醋酸外)...................... 80
表六(b)、洋蔥製品的有機酸含量比較(除醋酸外)...................... 81
附錄圖表
附圖一、所雷亞系統.................................................................... 91
附圖二、半連續式發酵法............................................................ 92
附圖三、具有香氣回收設備的發酵槽........................................ 93
附圖四、酒精及醋酸對醋酸菌生長的影響及最適培養濃度.... 94
附圖五、三階段添酒的醋酸發酵培養法.................................... 95
附表一、有效發表之醋酸菌種.................................................... 96
附表二、洋蔥汁、洋蔥醋及其他食醋的有機酸含量比較.......... 98
附表三、洋蔥醋與其他食醋之礦物質、總胺基酸及總有機酸
含量比較......................................................................
99

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