臺灣博碩士論文加值系統

本實驗以HPLC法探討八株乳酸菌包括Bifidobacterium longum B6和 15708；Lactobacillus acidophilus N-1和4356；Lactobacillus bulgaricus 448和 449；Streptococcus thermophilus MC和 573合成葉酸之能力並以MTT assay來測定葉酸對細胞株 Intestine 407 (CCRC 60022) 提高細胞存活和抗氧化傷害之影響。B. longum B6顯示有最高的葉酸生成能力，而B. longum 15708也有不錯的葉酸生成能力，在37℃經發酵6小時後葉酸分別增加了3.5倍和2.4倍。而添加乳糖則可增加葉酸合成及菌數；添加氯化鈣則些微增加菌數，但減少葉酸的生成。在4℃儲存期間之變化，顯示儲存時間越久，菌數及葉酸的損失就越多。在葉酸對細胞株存活率的影響試驗中，發現隨著葉酸濃度的提高，Intestine 407細胞的存活率就越高；而添加葉酸濃度的提高對Intestine 407細胞受到以H2O2為氧化劑時的氧化傷害也有降低的效果，顯示葉酸對Intestine 407細胞的生長和抗氧化傷害扮演很重要的角色。

Eight strains of lactic acid bacteria including Bifidobacterium longum B6 and 15708, Lactobacillus acidophilus N-1 and 4356, Lactobacillus bulgaricus 448 and 449, and Streptococcus thermophilus MC and 573 were investigated for folate synthetic ability by high-performance liquid chromatographic method. The effect of folate on cell viability and anti-oxidative damage of Intestine 407 (CCRC 60022) was evaluated by MTT assay. B. longum B6 demonstrated the highest folate synthetic ability, and B. longum 15708 also produced folate well. These two strains increased 3.5 and 2.4 folds of folate, respectively, after 6h fermentation at 37℃. The addition of lactose increased the level of folate and viable cell counts. Calcium chloride increased the numbers of viable cell counts but decreased the level of folate. Folate levels and viable cell counts decreased significantly when stored at 4℃. In MTT assay, cell viability of Intestine 407 increased when the concentration of folic acid increased. And oxidative damage of H2O2 to Intestine 407 was reduced when the concentration of folic acid increased. This study showed that folic acid plays a significant role in cell viability and anti-oxidative damage of Intestine 407.

目錄
目錄 Ⅰ
圖次Ⅳ
表次Ⅴ
壹、文獻整理 1
一、前言 1
二、葉酸之簡介 2
(一) 發現 2
(二) 結構 2
(三) 特性 3
(四) 代謝與吸收 4
(五) 缺乏的影響 8
(六) 食物來源 9
(七) 葉酸建議攝取量 9
(八) 毒性 12
三、合成葉酸之腸道細菌 12
四、葉酸對疾病之預防 14
五、乳酸菌之簡介 18
(一)乳酸菌的分類 18
(二)乳酸菌與人體健康的關係 20
貳、乳酸菌合成葉酸能力之探討 23
一、中文摘要 23
二、英文摘要 24
三、前言 25
四、材料與方法 27
(一) 試驗菌株 及細胞株27
(二) 實驗藥品與器材 27
(三) 葉酸標準品製備 30
(四) Extraction buffer和Human plasma 的製備31
(五) 實驗項目及步驟 31
五、結果與討論 36
(一) 生長曲線試驗36
(二) 葉酸的HPLC層析圖 36
(三) 葉酸生成試驗 37
(四) 添加乳糖與氯化鈣之影響 44
(五) 儲存期間之菌數與葉酸之變化 46
(六) 添加葉酸至細胞株Intestine 407的影響 53
參、參考文獻 57
圖次
壹、文獻整理 1
圖一、各種葉酸型式之結構 5
圖二、攝取的葉酸在人類消化與吸收的過程 6
圖三、四氫葉酸於單碳代謝及傳遞時所扮演之角色 7
貳、乳酸菌合成葉酸能力之探討 23
圖一、乳酸菌生長曲線圖 38
圖二、各種葉酸在激發波長295 nm下
螢光吸收掃瞄圖，掃瞄範圍305-700 nm 39
圖三、HPLC系統之葉酸層析圖 41
圖四、不同培養基中葉酸之生成曲線圖 42
圖五、儲存在4℃期間葉酸的變化51
圖六、儲存在4℃期間菌數的變化52
表次
壹、文獻整理 1
表一、食品中葉酸的含量 10
表二、各種維生素之每日建議攝取量 11
表三、乳酸菌的型態、發酵型式及好氣條件 20
貳、乳酸菌合成葉酸能力之探討 23
表一、發酵乳中各種葉酸型式的含量 40
表二、培養0與6小時之葉酸生成之比較 43
表三、添加乳糖及氯化鈣對乳酸菌菌數之影響 (6hr及18hr) 47
表四、添加乳糖及氯化鈣對乳酸菌葉酸之影響 (6hr及18hr)
48
表五、發酵乳在4℃儲存葉酸之變化49
表六、發酵乳在4℃儲存菌數之變化50
表七、添加葉酸對細胞存活之影響55
表八、添加葉酸對受到過氧化氫傷害之細胞生長的影響56

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