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dc.contributor.authorPeng, Ya-Weien_US
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dc.description.abstractLactobacillus reuteri存在於自然環境中,且能定殖於健康動物,包括人類的腸道中。經研究證實其具有多項利於人體健康的益生功效,故被認可為益生菌。目前不論在研究或應用方面,均日益受到重視。但是因其在冷凍貯藏之耐受性差,所以在食品上的應用並不廣泛。細胞微膠囊化技術可以有效的保護菌株,來提升菌株對不利的環境的耐受性,以及在穿越腸胃道的過程中提供保護,從而提高其存活率與活性,以及加工處理與貯藏過程中的耐受性。本研究利用褐藻酸鈣來對Lactobacillus reuteri 進行細胞微膠囊化處理,同時分別添加不同濃度(1%、2%、3%)的果寡醣作為保護劑,再放置於不同凍結溫度下 (-20℃、-40℃、-60℃、-80℃、-196℃) 來進行貯藏,探討其對L. reuteri在冷凍貯藏中的保護效果,以提升菌株之存活率、活性與貯藏安定性,並比較不同處理間的差異。結果顯示,細胞微膠囊化能提高菌株在冷凍貯藏的存活率。在凍結貯藏溫度-60℃下,菌株的存活率較高。且果寡醣的添加對於微膠囊化菌株的死滅有降低的效果,添加2%果寡醣並在-60℃冷凍溫度貯藏下,可以獲得最小的死滅速率。而菌株β-半乳糖苷酶活性,與菌株存活率結果相類似,在添加2%果寡醣並在-60℃冷凍溫度貯藏,能保留高達50%的β-半乳糖苷酶活性。脂肪酸成分分析結果顯示,微膠囊化處理和冷凍貯藏溫度對菌株之脂肪酸變化有顯著的影響,但是果寡醣添加則沒有影響。由蛋白質分析結果可發現,經不同處理之菌株皆無法合成新的蛋白質。綜合以上結果得知,在冷凍貯藏過程中,在-60℃凍結貯藏溫度和2%果寡醣添加之L. reuteri處理,其冷凍耐受性較佳。zh_TW
dc.description.abstractLactobacillus reuteri was found in many natural environments, and could colonize the intestines of healthy animals, including human. It shows certain beneficial effects to human health and is recognized as a probiotic. It has led to a great deal of further studies in the researches and applications. However, its application in frozen foods is still not popular because of its poor freezing tolerance during frozen storage. The cell microencapsulation technique has been found to possess great potential for protecting probiotic bacteria, such as lactic acid bacteria, against adverse conditions in food and during passage through the gastrointestinal tract. It could therefore yield improved survival and activity of probiotic bacteria, and resistance against the conditions in food processing and storage. In this study, Ca-alginate was applied to L. reuteri for cell microencapsulation, and different concentrations (1%, 2%, 3%) of fructooligosaccharides (FOS) was added as cryoprotectants at the same time for the storage test at different frozen temperatures (-20℃, -40℃, -60℃, -80℃, -196℃) in order to study their protection effects on the enhancement of cell survival, activity and storage stability of L. reuteri during frozen storage and effects of different treatments were also compared. Results indicated that cell microencapsulation could increase the cell survival during frozen storage, and -60℃ frozen storage could obtain the higher survival among different frozen temperatures used. FOS addition could reduce cell death combined with cell microencapsulation. Addition of 2% FOS and stored in - 60℃ freezing temperature was found to be the best conditions among all the various treatments. The data of fatty acid analysis revealed that the microencapsulation and the freezing storage temperature had the remarkable influence to the fatty acid change, but FOS addition had no effect. According to the protein analysis, no new intracellular protein was found in L. reuteri with different treatments. Based on the above results, it could conclude that 2% FOS addition with -60℃ freezing temperature storage could obtain the better freeze-tolerance.en_US
dc.description.tableofcontents中文摘要 i 英文摘要 ii 目錄 iv 圖目錄 vii 表目錄 ix 前言 1 壹、 文獻整理 2 一、 益生菌(probiotics)簡介 2 (一) 益生菌之定義 2 (二) 益生菌之特性 2 (三) 益生菌之生理功效 4 (四) 洛德乳桿菌(Lactobacillus reuteri) 8 二、 益菌生(prebiotics)簡介 12 (一) 益菌生之定義 12 (二) 益菌生之生理功效 12 (三) 常見益菌生之種類 16 (四) 共生質(synbiotics) 16 三、 微生物之冷凍耐受性 19 (一) 冷凍對微生物之影響 19 (二) 提升微生物冷凍耐受性之方法 21 (三) 冷凍保護劑(cryoprotectant) 21 (四) 多醣類之冷凍保護機制 24 四、 果寡醣(fructooligosaccharides) 27 (一) 果寡醣之化學結構 27 (二) 果寡醣作為益菌生 27 (三) 果寡醣作為冷凍保護劑 27 五、 細胞固定化(cell immobilization) 29 (一) 細胞固定化之技術 29 (二) 微膠囊化技術(microencapsulation) 31 (三) 褐藻膠(alginate) 35 貳、 研究目的 38 參、 材料與方法 39 一、 試驗材料與儀器 39 (一) 材料 39 (二) 儀器 43 二、 試驗方法 43 (一) 實驗架構 43 (二) 實驗菌株的製備 43 (三) 實驗樣本之製備 45 (四) 凍結儲藏試驗 48 (五) ο-nitrophenyl-β-galactopyranoside酵素試驗 48 (六) 脂肪酸分析 48 (七) 蛋白質分析 49 (八) 電子顯微鏡觀察 55 (九) 統計分析 55 肆、 結果與討論 57 一、 菌株之存活率 57 (一) 微膠囊化處理對菌株存活率之影響. 57 (二) 不同濃度之果寡醣添加對菌株存活率之影響 61 (三) 微膠囊化且添加果寡醣對菌株存活率之影響 64 二、 菌株之活性分析 69 (一) ο-nitrophenyl-β-galactopyranoside酵素試驗 69 (二) 微膠囊化處理對β-半乳糖苷酶活性之影響. 69 (三) 不同濃度之果寡醣添加對β-半乳糖苷酶活性之影響 72 (四) 微膠囊化且添加果寡醣對菌株β-半乳糖苷酶活性之影響 74 三、 菌株之菌體成分分析 76 (一) 脂肪酸成分分析 76 (二) 蛋白質成分分析 83 (三) 電子顯微鏡觀察 86 伍、 結論 92 陸、 參考文獻 94zh_TW
dc.subjectLactobacillus reuterien_US
dc.subjectLactobacillus reuterizh_TW
dc.subjectfrozen storageen_US
dc.titleLactobacillus reuteri冷凍耐受性提升之探討: 細胞微膠囊化及果寡醣添加的利用zh_TW
dc.titleStudies on the Enhancement of the Freeze-Tolerance of Lactobacillus reuteri: the Application of Cell Microencapsulation and Fructooligosaccharides Additionen_US
dc.typeThesis and Dissertationzh_TW
item.openairetypeThesis and Dissertation-
item.fulltextno fulltext-
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