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研究生:李耀全
論文名稱:熱休克處理或冷休克處理對冷凍乾燥嗜酸乳酸桿菌存活率及貯藏安定性的影響
論文名稱(外文):Effect of heat shock or cold shock treatment on the survival and storage stability of freeze-dried lactobacillus acidophilus
指導教授:金安兒金安兒引用關係
學位類別:碩士
校院名稱:國立中興大學
系所名稱:食品科學系
學門:農業科學學門
學類:食品科學類
論文種類:學術論文
論文出版年:2004
畢業學年度:92
語文別:中文
論文頁數:111
中文關鍵詞:熱休克冷休克嗜酸乳酸桿菌冷凍乾燥凍結存活率β-半乳糖苷活性產酸能力
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冷凍乾燥廣泛地被使用於菌種的保存,但是由於其經過了凍結及在凍結狀態下行真空乾燥的操作,導致了大多數菌體細胞的死亡。嗜酸乳酸桿菌(L. acidophilus)具有多項有益人體的生理功能,在市售乾燥食品中常有添加,有助人體健康,但是在乾燥食品中之存活率及儲存安定性均差。不少研究報告指出,菌體具有對不利環境之抵抗性及適應性,而且在不利環境下菌體的存活率可以藉由壓迫反應(stress response)之誘導來促進。因此本研究藉由熱休克(50℃,30分鐘)或冷休克(10℃,3小時)來進行處理,探討其對嗜酸乳酸桿菌在冷凍乾燥中抵抗力的誘導效果,以提升所得乾燥菌株之存活率與儲藏安定性。結果顯示,熱休克的菌株在冷凍及冷凍乾燥後有較高的存活率,冷休克的影響則較不明顯。在貯藏試驗方面,貯藏溫度對貯存結果有顯著的影響,低溫貯藏下的冷凍乾燥菌株存活率較高。貯藏15週後,25℃下貯藏的菌株之菌數呈現出明顯下降的趨勢,不過經過熱休克處理後的菌株,存活率顯著高於其他的組別。比較β-galactosidase的活性可發現,熱休克處理的菌株在冷凍乾燥後β-galactosidase活性下降,顯示細胞膜破壞較少。產酸能力試驗顯示熱休克或冷休克後的菌株能改善凍結傷害所造成之產酸能力的降低。由SDS-PAGE結果可發現熱休克處理會誘導具環境壓力抗性之蛋白質的生成。冷凍乾燥後,嗜酸乳酸桿菌的掃描式電子顯微鏡照片中,熱休克或冷休克處理後菌體外觀較完整,殘存的菌數多。由此推論熱休克處理可提升冷凍乾燥L. acidophilus菌株之存活率與貯藏安定性。
壹、前言…………………………………………………………… 1
貳、文獻整理……………………………………………………… 3
一、乳酸菌介紹……………………………………………… 3
(一)乳酸菌的發現………………………………………. 3
(二)乳酸菌的定義與分類………………………………. 4
(三)乳酸菌的代謝特性…………………………………. 6
(四)乳酸菌為優質益生菌(probiotics)……….………..... 7
(五)乳酸菌在食品加工上的應用…….………………… 10
(六)乳酸菌對人體健康的影響………………….……… 18
(七)嗜酸乳酸桿菌(Lactobacillus acidophilus)………… 25
二、冷凍乾燥……………………………………………....... 28
(一)冷凍乾燥之起源……………………………………. 28
(二)冷凍乾燥定義和原理………………………………. 29
(三)冷凍乾燥之四階段…………………………………. 29
(四)冷凍乾燥設備………………………………………. 33
(五)冷凍乾燥之優缺點…………………………………. 35
(六)冷凍乾燥之產品……………………………………. 39
三、乳酸菌之冷凍乾燥………....…………………………… 41
(一)以冷凍乾燥保存菌株….……………………...……. 41
(二)冷凍乾燥對菌株之傷害……………………………. 41
(三)影響冷凍乾燥菌株貯藏存活率之因素……………. 42
(四)冷凍乾燥菌粉的回溶………………………………. 46
四、環境壓迫反應…………………………………………… 48
(一)熱休克與冷休克反應……………………………… 48
(二)交叉保護(cross protection)………………………… 49
(三)乳酸菌之環境壓迫反應…………………………… 49
參、研究目的……………………………………………………… 51
肆、材料與方法…………………………………………………… 52
一、試驗材料………………………………………………… 52
二、試驗方法………………………………………………… 56
伍、結果與討論…………………………………………………… 64
一、凍結及冷凍乾燥後菌株存活率的比較………………… 64
二、不同壓迫處理後菌數存活率的比較…………………… 68
三、冷凍乾燥後L. acidophilus的貯藏安定性……………… 69
四、貯藏溫度對冷凍乾燥L. acidophilus之貯藏安定性
的影響…………………………………………………...
72
五、β-galactosidase activity……………………………..…… 78
六、產酸能力試驗…………………………………………… 80
七、SDS膠體電泳…………………………………………… 87
八、冷凍乾燥菌株之掃描式電子顯微鏡觀察……………… 89
陸、結論…………………………………………………………… 94
柒、中文摘要……………………………………………………… 96
捌、英文摘要……………………………………………………… 98
玖、參考文獻……………………………………………………… 100
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