臺灣博碩士論文加值系統

培養比菲德氏菌會產生一些代謝性有機酸如乳酸與醋酸，這些有機酸會抑制比菲德氏菌的生長。目前並無較可靠快速的方法來偵測培養比菲德氏菌過程中所產生的有機酸與乳糖的代謝情形。本研究探討培養比菲德氏菌longum CCRC14634過程中氧化還原電位的變化。我們利用二種不同的培養基以及批次、饋料批次、循環批次式、透析式和連續式等五種培養方式，來探討醱酵比菲德氏菌過程中細胞密度、培養基代謝（乳糖、葡萄糖）、代謝性有機酸濃度（乳酸、醋酸）與氧化還原電位的關係。
本研究結果發現偵測氧化還原電位可以作為判斷醱酵比菲德氏菌醱酵狀況的依據。隨著醱酵過程中主要碳源的消耗，氧化還原電位逐漸降低。以循環批次式醱酵培養可以氧化還原電位變化作為控制參數，當氧化還原電位開始持平或上升時，移除 3/4 體積的培養液，並饋入 3/4 體積的新鮮培養基以進行下一循環批次醱酵之用。在厭氧培養條件下，培養於 MRS 和 WY 培養基中，分別得到平均約 1.9×109 CFU/ml和3.42×109 CFU/ml的比菲德氏菌。另外，以 MRS 培養基進行批次饋料醱酵和透析式以去除代謝性有機酸醱酵，可得菌體量分別為 5.2×109 CFU/ml 和 1.8×1010 CFU/ml。以 WY 培養基進行連續式醱酵培養時可得菌體量為 3.1×109 CFU/ml。

The lactic acid and acetic acid are produced in the fermentation of Bifidobacteria. The metabolic organic acids inhibit cell growth during fermentation of Bifidobacteria. There are no reliable methods to detect the inhibitory organic acids and lactose uptake during fermentation. The present study monitored the ORP changes during fermentation of Bifidobacteria longum CCRC 14634. The batch, fed-batch, cyclic batch, continuous and dialysed fermentation were performed to culture Bifidobacteria. Cell densities, uptake of carbon sources (glucose, lactose), metabolic organic acids, and ORP were detected during fermentation.
It was found that ORP was very close related to cell growth of Bifidobacteria. ORP decreased continuously during fermentation until exhausting of carbon source. We have performed six cycles of batch fermentation using ORP as control parameter. When ORP remained constant or increased, three fourth of the medium was removed, and the same volume of fresh medium was fed to the fermentor for each cycle. We have achieved 1.9×109 CFU/ml and 3.4×109 CFU/ml in average for cyclic batch fermentation in MRS and WY medium respectively. Furthermore, cell densities of 5.2×109 CFU/ml and 1.8×1010 CFU/ml were achieved by fed-batch and dialysed fermentation respectively in MRS medium. Continuous fermentation was performed using WY medium to achieve 3.1×109 CFU/m

摘要…………………………………………………………………I
英文摘要……………………………………………………………II
目錄…………………………………………………………………III
表和圖目錄…………………………………………………………V
第一章 前言………………………………………………………1
第二章 文獻回顧…………………………………………………2
2.1 雙歧桿菌之分類、型態和分布情形………………………2
2.2 雙歧桿菌之代謝途徑………………………………………3
2.3 雙歧桿菌之促進因子………………………………………8
2.4 雙歧桿菌和人體健康的關係………………………………10
2.4.1 維持人體常到菌從之平衡……………………………10
2.4.2 活化免疫系統…………………………………………12
2.4.3 促進蛋白質分解………………………………………14
2.4.4 抗癌作用及降低肝臟負擔……………………………14
2.4.5 促進乳酸之代謝………………………………………16
2.4.6 合成維生素 B 群………………………………………16
2.4.7 改善乳糖不耐症、腹瀉和便秘………………………16
2.4.8 抗氧化性………………………………………………17
2.4.9 降低血液中膽固醇和降低血壓………………………17
2.5 雙歧桿菌的耐氧性、耐酸性和耐膽鹽性…………………18
2.6 雙歧桿菌在食品界的應用…………………………………22
2.7 乳清和乳糖…………………………………………………22
2.8 氧化還原電位的偵測………………………………………27
第三章 材料與方法…………………………………………………29
3.1 培養基成分…………………………………………………29
3.2 儀器和設備…………………………………………………30
3.3 實驗方法……………………………………………………32
3.3.1 菌種培養和保存………………………………………32
3.3.2 醱酵培養………………………………………………32
3.3.2.1 批次醱酵…………………………………………33
3.3.2.2 批次饋料醱酵……………………………………33
3.3.2.3 循環批次式醱酵培養……………………………33
3.3.2.4 透析式醱酵培養…………………………………35
3.3.2.5 連續式醱酵培養…………………………………35
3.4 結果分析……………………………………………………37
第四章 結果與討論…………………………………………………39
4.1 批次醱酵……………………………………………………39
4.2 批次饋料醱酵………………………………………………44
4.3 循環批次式醱酵培養………………………………………46
4.4 透析式醱酵培養……………………………………………51
4.5 連續式醱酵培養……………………………………………53
第五章 結論…………………………………………………………55
參考文獻………………………………………………………………56
附錄……………………………………………………………………63

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