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標題: Lactobacillus 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
Studies on the Enhancement of the Freeze-Tolerance of Lactobacillus reuteri: the Application of Cell Microencapsulation and Fructooligosaccharides Addition
作者: 彭雅薇
Peng, Ya-Wei
關鍵字: Lactobacillus reuteri;Lactobacillus reuteri;frozen storage;microencapsulation;cryoprotectant;fructooligosaccharides;freeze-tolerance;冷凍貯藏;微膠囊化;冷凍保護劑;果寡醣;冷凍耐受性
出版社: 食品暨應用生物科技學系所
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Lactobacillus 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。

Lactobacillus 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.
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