本 研 究 以 褐 藻 酸 鈣(Ca-alginate)和幾丁聚醣-褐藻酸鈣複合材質 (Chitosan-Ca-alginate complex material) 來 包 埋 固 定 化 Lactobacillus reuteri，探討經模擬人體胃腸道條件作用後，於模擬結腸液中溶離釋出
後的菌體存活率，再以 Caco-2 細胞之生體外試驗模式探討釋出之 L.
reuteri 對腸道細胞吸附性、對 Listeria monocytogenes 的拮抗性、益生菌
之安全性等多方面基礎益生特性，並檢測 β-galactosidase 活性以了解模 擬胃腸道環境條件對L. reuteri 菌體細胞膜之影響。總結研究結果如下：
(1) 菌體於模擬胃腸道條件作用之耐受性試驗結果顯示，固定化能提高 L. reuteri 菌株經模擬人體胃腸道條件作用後的存活率，其中以幾丁聚
醣-褐藻酸鈣複合材質保護菌體的結果較佳。所有試驗條件中，以 pH 3 酸液作用 3 小時，再以 0.1%之膽鹽作用 3 小時，最後於模擬結腸液
中作用1 小時的存活菌數與存活率最高。
(2) L. reuteri 經相同 pH 值之酸液作用後，隨著膽鹽濃度增加，β-半乳糖 苷酶的活性也隨之增加，顯示菌體細胞膜之受損程度也隨之增加，而 菌體經包埋固定化則可顯著減少酸與膽鹽對菌體細胞膜的傷害。
(3) 菌體經 pH 3 酸液、0.1%膽鹽及模擬結腸液連續作用後仍具有吸附能 力，經過固定化的二試驗組其吸附率和游離態菌體相比是顯著較高
的，其中以幾丁聚醣-褐藻酸鈣複合材質組之吸附率最高(0.524%)。由 結果推測包埋的菌體因固定化而減少菌體表面蛋白質成分之吸附因 子受到酸液及膽鹽的損害，因此固定化能保護菌體並維持其吸附能 力。
(4) 經模擬胃腸道條件作用後的 L. reuteri 仍保有拮抗病原菌之能力且可 有效減少吸附於Caco-2 細胞上的 L. monocytogenes 數目，其中經包埋 後釋出的菌體其拮抗能力優於游離態菌體者。
(5) 侵入性試驗的結果顯示包埋後釋出的菌株具有安全性，可推測本研究 所使用的菌株是具有安全性，沒有侵襲或感染人體腸道細胞的危險 性。
(6) 以酸、膽鹽與模擬結腸液之連續作用模式來模擬胃腸道環境條件，相 當接近人體之生體內環境，以之探討經上述條件作用後之益生菌的益 生特性為適當可行之生體外試驗評估模式。
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