探討以Agaricus blazei Murrill、Lactobacillus buchneri二階段發酵Camellia oleifera及Camellia sinensis生產高GABA發酵茶

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吳越安(Yue-An Wu) 查詢紙本館藏

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化學工程與材料工程學系

論文名稱

探討以Agaricus blazei Murrill、Lactobacillus buchneri二階段發酵Camellia oleifera及Camellia sinensis生產高GABA發酵茶
(High GABA Fermented Tea of Camellia sinensis and Camellia oleifera by Lactobacillus buchneri and Agaricus blazei Murill using Two-Stage Fermentation)

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摘要(中)

飲茶文化發源自中國，已有上千年歷史。茶文化影響範圍不單受限於亞洲
地區，十七世紀初荷蘭、葡萄牙兩國透過海上貿易，將中國茶文化帶入西方世
界，引發一股巨大的飲茶風潮。直至今日，茶飲品受普羅大眾所喜愛，在飲品
圈內需求量僅次於水。已有許多研究證實茶對於人體健康保健具有不容小覷的
功效，如增強身體免疫力、降低膽固醇等。茶葉中含有大量多酚類化合物，它
的存在讓茶有著很優秀的抗氧化活性。 γ-氨基丁酸 (GABA) ——一種廣泛存
在於動植物中的胺基酸，在人體中存在於大腦及神經系統，主要功能為鎮靜神
經及調適情緒，近年來人們透過 GABA 改善失眠症狀。在這項研究中，利用
巴西蘑菇 (Agaricus blazei Murill) TITC1 以及布氏乳桿菌 (Lactobacillus
buchneri) BCE119151 做為發酵之菌種，在本實驗室過往研究所得之最適化培
養基條件下，對綠茶 (Camellia sinensis) 及油茶 (Camellia oleifera) 萃取液進
行二階段發酵，目的在於生產高 GABA 含量之發酵茶飲品並探討影響 GABA
產量的操作條件及細節。巴西蘑菇中富含多種胺基酸成分，又以其中的麩胺酸
(Glutamic acid) 最為重要，其做為 γ-氨基丁酸的前驅物能夠刺激布氏乳桿菌中
的 GABA 轉化酶生產高濃度 GABA。在這些菌種之二階段發酵下，使得後發
酵茶飲品能夠有更高的保健價值。
在第一階段巴西蘑菇發酵過程中，發現培養基中茶粉固體存在與否會影響
GABA 產物濃度。將實驗過程中茶葉萃取液和茶粉固體分離後做為培養基使
用，其最終 GABA 產物濃度高於含茶粉固體之培養基所得產物濃度。在第二
階段布氏乳桿菌發酵過程中，試驗巴西蘑菇之存活與否對最終 GABA 產物濃
度產生影響，實驗結果為第一階段之巴西蘑菇在第二階段布氏乳桿菌發酵過程
中無論是活菌抑或是死菌其結果無明顯差異。比較以各式茶作為底物，分別為
油茶、綠茶、紅茶和烏龍茶，分別得出最終 GABA 產量為 25.94 g/L、28.62 g/L、
ii
29.18 g/L 和 29.55 g/L。若將咖啡因也考慮進來，比較每克 GABA 所含每毫克咖
啡因 (YC/G)，油茶將是首選。

摘要(英)

The culture of drinking tea originated in China, it has history of thousands of years.
The influence of tea culture is not only in the Asia, but also in the Europe. In the early
17th century, the Netherlands and Portugal brought Chinese tea culture to the Western
world through trade, triggering a huge wave of tea drinking. Nowadays, drinks of tea
are loved by the general public, and its consumption is merely second to water. Many
studies have confirmed that tea has various health effects on the human body, such as
enhancing the body′s immunity and lowering cholesterol. Tea contains a large amount
of polyphenolic compounds, and it makes tea have excellent antioxidant activity. γaminobutyric acid (GABA)—an amino acid that widely exists in animals and plants. It
exists in the brain and nervous system in the human body. Its main function is to calm
the nerves and adjust emotions. Recently, people have used GABA to improve insomnia.
In this study, Agaricus blazei Murill TITC1 and Lactobacillus buchneri BCE119151
were used as our fermentation strains. Under the optimal medium conditions obtained
in previous research in our laboratory, the extracts of green tea (Camellia sinensis) and
oil camellia (Camellia oleifera) are used into two-stage fermentation, the purpose is to
produce fermented tea with high GABA content and also to explore the operating
conditions and details that affect GABA production. Agaricus blazei Murill are rich in
different amino acids. One of them, glutamic acid is the most important. As a precursor
of γ-aminobutyric acid, it can stimulate the glutamic acid decarboxylase, GAD in
Lactobacillus buchneri to produce a lot of GABA. Under the two-stage fermentation of
these strains, the fermented tea can be more beneficial for human body.
During the first stage of Agaricus blazei Murill fermentation, it was found that the
presence or absence of tea powder in the culture affected the GABA concentration in
iv
final product. During the experiment, the tea extract and tea powder were separated and
only used extract into culture. The final GABA concentration of product was higher than
the product that was from the culture containing tea powder. In the second stage of
fermentation of Lactobacillus buchneri, whether the survival of Agaricus blazei Murill
affected the concentration of final GABA products was tested. There was no significant
difference in the results of the two conditions. Tried to compare with oiltea Camellia、
green tea、black tea and oolong tea for fermentation culture, finally I get the final GABA
concentration for them with 25.94 g/L、28.62 g/L、29.18 g/L 和 29.55 g/L. Considering
the caffeine by YC/G (Caffeine per milligram / GABA per gram), oiltea Camellia will be
the best fermentation culture.

關鍵字(中)

★ 巴西蘑菇
★ 布氏乳桿菌
★ 油茶
★ 茶葉
★ γ-胺基丁酸

關鍵字(英)

★ Agaricus blazei Murrill
★ Lactobacillus buchneri
★ Camellia oleifera
★ Camellia sinensis
★ γ-Aminobutyric acid

論文目次

摘要 i
Abstract iii
致謝 v
目錄 vi
圖目錄 ix
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 巴西蘑菇 (Agaricus blazei Murrill) 3
2-1-1 巴西蘑菇的基本介紹 3
2-1-2 巴西蘑菇的組成成分 5
2-1-3 巴西蘑菇的生物活性物質 6
2-1-4 巴西蘑菇的醫學價值 12
2-2 乳酸菌 (Lactic acid bacteria, LAB) 13
2-2-1乳酸菌的基本介紹 13
2-2-2 乳酸菌的生物活性物質 16
2-2-3 布氏乳桿菌 (Lactobacillus buchneri) 19
2-2-4 乳酸菌發酵飲品 19
2-3 油茶 (Camellia oleifera) 20
2-3-1 油茶的基本介紹 20
2-3-2 油茶的成分介紹 21
2-4 茶 (Camellia sinensis) 23
2-4-1 茶樹的基本介紹 23
2-4-2 茶 (Tea) 24
2-4-3 康普茶(Kombucha) 26
三、材料與方法 29
3-1 實驗架構 29
3-2 實驗材料 30
3-2-1 實驗菌株 30
3-2-2 實驗樣品 31
3-2-3 實驗藥品 31
3-2-4 實驗儀器與設備 33
3-3 實驗方法 35
3-3-1 巴西蘑菇菌株保存 35
3-3-2 巴西蘑菇菌株之液態搖瓶發酵 35
3-3-3 布氏乳桿菌菌株短期保存與活化 36
3-3-4 布氏乳桿菌菌株之液態種瓶培養 37
3-3-5 第一階段之巴西蘑菇搖瓶發酵 38
3-3-6 第二階段之布氏乳桿菌厭氧發酵 38
3-4 分析方法 39
3-4-1 還原醣濃度分析 39
3-4-2 GABA含量和MSG殘量分析 41
3-4-3 總可溶醣濃度分析 44
3-4-4 兒茶素濃度分析 45
3-4-5 茶色素含量分析 47
3-4-6 總多酚含量分析 49
3-4-7 咖啡因含量分析 50
四、結果與討論 51
4-1 巴西蘑菇、布氏乳桿菌二階段發酵之操作優化 51
4-1-1 發酵培養基中茶粉渣存在對發酵產物之影響 51
4-1-2 第二階段培養基中巴西蘑菇存活與否對發酵產物之影響 52
4-2 各種不同茶底物之發酵結果 54
4-2-1 各種不同發酵茶之最終GABA產量 54
4-2-2 各種不同發酵茶之最終MSG剩餘量 55
4-2-3 各種不同發酵茶之最終咖啡因含量 56
4-3 各種不同發酵茶之生物活性物質含量 58
4-3-1 各種不同發酵茶之總兒茶素含量 58
4-3-2 各種不同發酵茶之茶色素含量 59
4-3-3 各種不同發酵茶之總多酚含量 60
4-4 各種不同發酵茶之菌種活性變化 61
4-5 各發酵茶之發酵結果比較 61
4-6 利用茶渣為底物之發酵茶 63
五、結論 64
參考文獻 66

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指導教授

徐敬衡(Chin-Hang Shu)

審核日期

2023-7-22

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