臺灣博碩士論文加值系統

本研究以1% Hylon VII 抗性澱粉作為益生質，以蕃茄汁培養基培養乳酸菌，添加蔗糖 (S) 及麥芽糊精 (M) 做為冷凍保護劑進行冷凍乾燥處理製備乳酸菌微膠囊。從食品分離之乳酸菌分離株PM206 及 PM212在五種不同濃度10％M、20％M、5％S、10％S 及 5％M+5％S之冷凍保護劑進行冷凍乾燥。實驗結果顯示，PM206 及 PM212 分離株分別添加五種不同濃度保護劑以凍乾處理後所得到之乳酸菌微膠囊中菌數皆為109 CFU/g，PM212在 5％S 於凍乾後則可達1010 CFU/g，保護效果最佳。凍乾製備之乳酸菌微膠囊於7℃ 儲藏 180 天期間之菌數皆維持在109 ~ 1010 CFU/g，添加麥芽糊精 (10％M 及20％M) 之乳酸菌微膠囊相較於添加蔗糖 (5％S、10％S 及 5％M+5％S) 之乳酸菌微膠囊乾燥效果較佳，尤其是20%M 之乳酸菌微膠囊水分含量低於2.0 %，水活性低於0.1，乾燥效果最佳。進一步探討不同濃度保護劑之乳酸菌微膠囊於7℃ 保存 180 天期間對酸與膽鹽的耐受程度以及利用Caco-2細胞分析乳酸菌附著能力與乳酸菌對癌細胞活性之影響。實驗結果顯示，添加1% Hylon VII及5%S 和20%M之乳酸菌微膠囊，保存180天期間於模擬胃腸道試驗中對酸及膽鹽具有較良好的耐受性，兩株乳酸菌數皆維持在106 CFU/g以上。添加1% Hylon VII 及保護劑之 PM206 及 PM212 微膠囊對 Caco-2 細胞具有較佳之附著能力並抑制Caco-2細胞活性。綜合上述，添加1% Hylon VII及20%M之 PM206 及 PM212 微膠囊有助於乳酸菌菌株於儲藏期間之存活、提升乳酸菌株耐酸耐膽鹽之能力，且可以增加乳酸菌株對 Caco-2細胞活性的抑制。

In this study, we use 1% Hylon VII resistant starch as a prebiotic substance and culture lactic acid bacteria (LAB) in tomato juice as medium. In the freeze-dry process to prepare lactic acid bacteria microcapsules which we add the Sucrose (S) and Maltodextrin (M) as protective agent. The LAB, PM206 and PM212 isolated from foods, were treated five concentrations of M and S which was 10% M, 20% M, 5% S, 10% S and 5% M + 5% S to protect the damage in the freeze dry process. The results show that the numbers of PM206 and PM212 with five concentrations of protective agent was up to 109 CFU/g except PM212 was 1010 CFU/g. Freeze-dried LAB microcapsules was storage at 7℃for 180 days and showed LAB were still maintained at 109 ~ 1010 CFU/g. Adding Maltodextrin (10%M and 20%M) showed the better result in moisture content test and water activity test, especially the LAB with 20%M that moisture content was 2.0% and water activity was below 0.1. Furthermore we studied the survival rate Caco-2 cell that co-culture with PM206 and PM212 which storage at 7℃for 180 days and treated with acid and bile. The result showed that the numbers of LAB were all maintained at 106 CFU / g or even higher and LAB microcapsules with 1% Hylon VII and added 5%S and 20%M showed the better adhered activity to inhibited the growth of Caco-2 cells. In the conclusion, the PM206 and PM212 microcapsule with 1% Hylon VII and the protecting agent showed the have better adhesion activity to Caco-2 cells and inhibited the growth of Caco-2 cells.

中文摘要 I
Abstract II
目錄 III
表目錄 VI
圖目錄 VIII
壹、前言 1
貳、文獻整理 2
一、益生菌 2
(一) 益生菌之定義 2
(二)益生菌之特性 2
二、乳酸菌 3
(一) 乳酸菌之定義 3
(二) 乳酸菌之類型 3
(三) 乳酸菌之應用 4
(四) 乳酸菌對人體健康之益處 5
(五) 乳酸菌抵抗胃腸道之耐受性 7
三、益生質 8
(一) 益生質之定義 8
(二) 抗性澱粉之定義 8
(三) 抗性澱粉的種類 8
(四) 抗性澱粉的功用 9
(五) 抗性澱粉作為益生質之特性 10
四、冷凍乾燥 11
(一) 冷凍乾燥的原理 11
(二) 冷凍乾燥對乳酸菌的影響 11
(三) 冷凍乾燥之保護劑 12
(四)凍乾產品之儲藏安定性 13
五、乳酸菌抑制結腸直腸癌細胞之研究 13
(一) 細胞培養目的 13
(二) 常用的腸道細胞株 13
參、材料與方法 15
一、實驗架構 15
二、實驗材料 16
(一) 實驗菌株 16
(二) 抗性澱粉 16
(三) 培養基 17
(四) 藥品試劑 20
(五) 實驗耗材 21
(六) 實驗設備 22
三、實驗方法 23
(一) 乳酸菌保存與活化 23
(二) 乳酸菌於抗性澱粉存活性之分析 23
(三)冷凍乾燥之微膠囊化 23
(四)乳酸菌微囊粒徑分析 24
(五) 乳酸菌微囊色澤分析 24
(六)乳酸菌微膠囊水分含量 24
(七) 乳酸菌微膠囊水活性 25
(八) 乳酸菌微膠囊儲藏性試驗 25
(九) 凍乾之乳酸菌微膠囊菌數計數 25
(十)乳酸菌耐酸耐膽鹽試驗 25
(十一) 乳酸菌附著Caco-2細胞試驗 26
(十二)乳酸菌對Caco-2細胞活性分析 27
肆、結果與討論 28
一、乳酸菌對抗性澱粉之利用性分析 29
二、抗性澱粉及保護劑對乳酸菌冷凍乾燥之影響 29
三、乳酸菌微膠囊於儲藏期間之菌數變化 30
四、乳酸菌微膠囊之耐酸耐膽鹽試驗 31
五、乳酸菌微膠囊對Caco-2細胞之吸附能力 34
六、乳酸菌微膠囊對Caco-2細胞活性之影響 35
七、乳酸菌微膠囊之水分含量 35
八、乳酸菌微膠囊之水活性 36
九、乳酸菌微膠囊之粒徑分析 36
十、乳酸菌微膠囊之色澤分析 37
伍、結論 38
陸、圖表 39
柒、參考文獻 74

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