探討以Lactobacillus buchneri發酵巴西蘑菇並產生 γ-氨基丁酸之研究

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彭文正(Wen-Zheng Peng) 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Lactobacillus buchneri發酵巴西蘑菇並產生 γ-氨基丁酸之研究
(Evaluation that produce γ-aminobutyric acid during Agaricus blazei Murrill fermentation by Lactobacillus buchneri BCE11915)

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至系統瀏覽論文 (2025-9-1以後開放)

摘要(中)

長久以來，食用蘑菇因其營養價值和藥用特性而深受人們的消費與讚賞。巴西蘑菇（Agaricus blazei Murill）是一種源自於巴西東南部山區的傘菌屬真菌，因具有許多活性物質而廣受大家注目，尤其是抗腫瘤多醣體，在諸多文獻中都進行廣泛深入的研究。但由於目前缺乏巴西蘑菇與乳酸菌的相關研究，因此本論文目的為將自行分離篩選之乳酸菌與巴西蘑菇子實體及菌絲體進行發酵，並產生高含量γ-氨基丁酸（GABA）之發酵(液)產品，藉以評估巴西蘑菇作為發酵培養基之發展潛力。
在自然發酵的巴西蘑菇樣品液中，我們成功篩選出BCE119151乳酸菌株，並顯示出最高GABA產生能力。根據革蘭氏染色、生化試驗及16S rDNA序列測定結果，表明BCE119151為Lactobacillus buchneri(布氏乳桿菌)。在結合子實體發酵部分，使用BCE119151可成功發酵巴西蘑菇和黃豆粉混合物（ABSFS）。根據實驗結果，於初始pH 6.0、37℃ 操作條件下，可產生27.34 g/L GABA產量、 59.6% GABA轉化率；在未添加Monosodium glutamate (MSG)情況下，也可成功轉化出214 mg/L的GABA含量。在結合菌絲體發酵部分，利用豆漿作為第一階段巴西蘑菇菌絲體培養基並進行搖瓶發酵，再以連續式操作接著第二階段乳酸菌發酵，成功地在最大菌絲重情況下，發酵轉化出24.54 g/L的GABA含量、其GABA轉化率為62.6%；未添加MSG情況下，也可於最大麩胺酸含量下轉化出149 mg/L的GABA含量。綜合上述結果，巴西蘑菇與L. buchneri BCE119151結合進行發酵，可發展出具有高營養價值和功能特性之GABA發酵產品，並應用於相關飲品及其保健食品開發。

摘要(英)

For many years, edible mushrooms have been consumed and appreciated by their nutritional value, and medicinal properties. Agaricus blazei Murrill (ABM) is a mushroom native from Brazil largely studied due to its bioactive compounds, particularly polysaccharides. The aim of this work was to evaluate the potential of ABM as basal medium for the lactic acid fermentation of high γ-aminobutyric acid (GABA)-producing strain which was isolated from fermented edible mushroom. Owing to the lack of a detailed study on fermented ABM with lactic acid bacteria (LAB), the current study was employed to exploit and optimize the fermentation condition for the production of GABA with LAB.
LAB that accumulated GABA in culture medium were screened to identify strains with high GABA-producing ability. One strain, BCE119151, showed the highest GABA-producing ability among the screened strains. BCE119151 was identified as Lactobacillus buchneri based on Gram-staining, metabolic characteristics, and 16S rDNA sequence determination. ABM fruiting bodies powder and soybean flour mixture (ABSF) was successfully fermented using BCE119151. According to our results, the fermentation was optimized using the ABSF mixture (ABMP: soybean flour=2.5:1), with BCE119151, at an initial pH of 6.0, the incubation temperature at 37 degrees C for 7 days. Under these conditions, BCE119151 produced GABA at a concentration of 27.34 g/L with a 59.6% GABA conversion rate, it was also successfully produced 214 mg/L of GABA content without added MSG.
In addition, the first soymilk mycelial fermantation with BCE119151, 24.54 g/L of GABA was produced from the mycelial soymilk medium in maximum of TITC1 biomass and with a 62.6% GABA conversion rate.It was also produced 149 mg/L of GABA content without added MSG.The results suggested that the Agaricus blazei Murrill and L. buchneri BCE119151 possessed a prospect to be applied in fermented plant beverages and other health products with high nutritive values and functional properties.

關鍵字(中)

★ 巴西蘑菇
★ 菌絲體
★ 乳酸菌
★ 豆漿
★ GABA

關鍵字(英)

論文目次

摘要(Abstract) i
致謝 iii
目錄 iv
圖目錄 viii
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 Murill) 3
2-1-1巴西蘑菇的基本介紹 3
2-1-2巴西蘑菇的成分組成 3
2-1-3巴西蘑菇的生物活性物質 8
2-1-4巴西蘑菇的藥理研究 14
2-2 乳酸菌(Lactic acid bacteria) 16
2-2-1 乳酸菌的基本介紹 16
2-2-2 益生菌的基本介紹 20
2-2-3 Lactobacillus buchneri的介紹 21
2-2-4 乳酸菌發酵飲品介紹 22
2-3 乳酸菌產生的生物活性物質 25
2-3-1 γ-氨基丁酸(GABA) 25
2-3-2 硫代葡萄糖苷(Glucosinolates) 27
2-3-3 共軛亞麻油酸(Conjugated linoleic acid，CLA) 29
2-4 影響發酵工程的物化因子 31
2-4-1 培養基組成 31
2-4-2 pH值 33
2-4-3 溫度 34
2-4-4 攪拌速率 34
三、材料與方法 35
3-1 實驗架構 35
3-2 實驗材料 37
3-2-1 實驗菌株 37
3-2-2 實驗樣品 37
3-2-3 實驗藥品 38
3-2-4 實驗儀器與設備 40
3-3 實驗方法 42
3-3-1 乳酸菌分離純化 42
3-3-2 高GABA乳酸菌株篩選 43
3-3-3 高GABA乳酸菌株之相關特性測試 43
3-3-4 Lactobacillus buchneri BCE119151菌株保存及活化 44
3-3-5 Lactobacillus buchneri BCE119151種菌源製備 45
3-3-6 巴西蘑菇子實體發酵最適化條件探討 46
3-3-7 巴西蘑菇子實體發酵-操作條件探討 47
3-3-8 巴西蘑菇子實體發酵-GABA產量最適化探討 48
3-3-9 巴西蘑菇子實體發酵-發酵優化條件探討 48
3-3-10巴西蘑菇子實體發酵-不添加MSG 48
3-3-11 巴西蘑菇菌株保存及活化甘油保存 49
3-3-12 巴西蘑菇菌株液態搖瓶發酵 49
3-3-13 巴西蘑菇菌絲體發酵-第一階段搖瓶發酵 51
3-3-14 巴西蘑菇菌絲體發酵-第二階段乳酸菌厭氧發酵 52
3-4 分析方法 54
3-4-1 生化特性試驗 54
3-4-2 乳酸菌菌體濃度分析 55
3-4-3 pH值分析 55
3-4-4 菌落數分析(平板菌落計數法) 55
3-4-5 菌絲體乾重分析 56
3-4-6 還原糖濃度分析 56
3-4-7 總可溶性糖濃度分析 57
3-4-8 多醣體濃度分析 58
3-4-9 GABA及MSG含量分析 59
3-4-10 游離麩胺酸(Glutamic acid)分析 61
3-4-11 GAD酵素活性分析 63
四、結果與討論 65
4-1 乳酸菌分離純化 65
4-2 高GABA乳酸菌株篩選 66
4-3 高GABA乳酸菌株之相關特性測試 67
4-3-1 生化特性試驗及菌種鑑定 67
4-3-2 生長曲線及GABA發酵動力曲線 68
4-4 巴西蘑菇子實體發酵最適化條件探討 70
4-4-1 額外高蛋白營養源添加 70
4-4-2 調整A/S ratio組成(ABMP / Soy flour) 72
4-4-3 最適碳源添加 74
4-4-4 巴西蘑菇子實體發酵最適化條件之結論 77
4-5 巴西蘑菇子實體發酵-優化條件探討 79
4-6 巴西蘑菇子實體發酵-操作條件探討 81
4-6-1 不同溫度及pH值對發酵結果之影響 81
4-6-2 不同培養溫度對pH值及可溶性糖消耗量之影響 83
4-6-3 不同培養溫度對MSG及GABA含量之影響 85
4-6-4 不同培養溫度之發酵動力曲線 86
4-6-5 巴西蘑菇子實體發酵-操作條件探討之結論 88
4-7 巴西蘑菇子實體發酵-GABA產量最適化探討 90
4-8 巴西蘑菇子實體發酵(不添加MSG) 93
4-9 巴西蘑菇(Agaricus blazei Murrill)菌株液態搖瓶發酵 95
4-9-1 液態搖瓶發酵之游離麩胺酸含量探討 95
4-10 巴西蘑菇菌絲體發酵-第一階段搖瓶發酵 97
4-10-1 最適豆漿培養基固液比 97
4-10-1 巴西蘑菇菌絲體之豆漿培養基發酵動力曲線 98
4-11 巴西蘑菇菌絲體發酵-第二階段乳酸菌厭氧發酵 100
4-11-1 最大菌絲體重下進行乳酸菌發酵 100
4-11-2 最大粗多醣體含量下進行乳酸菌發酵 101
4-11-3 最大游離麩胺酸含量下進行乳酸菌發酵(不添加MSG)
102
4-12 巴西蘑菇子實體及菌絲體發酵之結論 103
五、結論 105
參考文獻 107
附錄 114

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指導教授

徐敬衡

審核日期

2020-8-5

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