臺灣博碩士論文加值系統

本研究探討乳酸菌液態發酵及生產活菌粉末之產程條件。針對菌株、培養基和乾燥程序進行研究。本研究主要研究目的有三：首先改善乳酸菌液態培養的牛乳培養基配方，提高以牛乳為主培養基的乳酸菌發酵液之菌體濃度；其次將改善後培養基應用於80L乳酸菌批次發酵，並建立一完整之發酵流程；最後將80L乳酸菌批次發酵的發酵液利用低溫負壓式流動床噴霧造粒乾燥機進行噴霧乾燥製造粉劑，測試其噴霧乾燥流程的操作條件。本研究的研究菌株有：Lactobacillus acidophilus、Bifidobacterium bifidum、Lactobacillus sporogenes 等三株乳酸菌，其中Lactobacillus sporogenes 是一會形成內生孢子的乳酸菌，是本文較重視的研究菌種。
在液態培養基的配方改善研究方面，Lactobacillus acidophilus最高菌體濃度為4.78 ×109 CFU/mL（30 ℃, 10% milk powder, 1600 U/mL bromelain, add 1 % glucose at 12hr, ends at 24hr），Bifidobacterium bifidum最高菌體濃度為5.23×109 CFU/mL（28 ℃, 8 % milk powder, 1600 U/mL bromelain, add 1 % glucose at 12 hr, ends at 24 hr），Lactobacillus sporogenes最高菌體濃度4.38×1010 CFU/mL（33 ℃, 8 % milk powder, 1600 U/mL bromelain, add 1 % glucose at 12 hr, ends at 24 hr）。
在80L乳酸菌批次發酵方面，Lactobacillus acidophilus最高菌體濃度為2.27×109 CFU/mL（30 ℃, 10 % milk powder, 1600 U/mL bromelain, add 1 % glucose at 12 hr, stir 10 sec/12hr, ends at 24 hr），Bifidobacterium bifidum最高菌體濃度為1.31×109 CFU/mL（28 ℃, 8 % milk powder, 1600 U/mL bromelain, add 1 % glucose at 12 hr, stir 10 sec/12hr, ends at 24 hr），Lactobacillus sporogenes最高菌體濃度4.27×109 CFU/mL（33 ℃, 8 % milk powder, 1600 U/mL bromelain, add 1 % glucose at 12 hr, stir 10 sec/12hr, ends at 24 hr）。
在乳酸菌粉劑的製程上，現階段可以全程在35℃ 下36 小時內將80L 乳酸菌發酵液噴霧乾燥成8Kg 108 CFU/g 的Lactobacillus acidophilus 和Bifidobacterium bifidum 粉末，及8Kg 109 CFU/g 的Lactobacillus sporogenes 粉末；而本研究亦探討流程和設備上的缺失，未來若能改善這些問題，將會提升本系統的效率和效果，使本系統更趨於完善。

This research studied the submerged fermentation, strains, medium composition and drying process for the production of lactic acid bacteria. The application of a reduced pressure, low temperature fluidized drying system on the powdered product was also studied. Three strains: Lactobacillus acidophilus, Bifidobacterium bifidum and Lactobacillus sporogenes were used in this study. One of which, Lactobacillus sporogenes can form endospore and is believed to be advantageous to its survival in the human digestive system, is emphasized in this research.

Using improved cultural medium flor flask culture, the cell concentration of Lactobacillus acidophilus was 4.78 ×109 CFU/mL (30℃, 10% milk powder, 1600U/mL bromelain, add 1% glucose at 12 hr, harvest at 24 hr); while that for Bifidobacterium bifidum was 5.23×109 CFU/mL (28℃, 8% milk powder, 1600U/mL bromelain, add 1% glucose at 12 hr, ends at 24 hr). For Lactobacillus sporogenes, the cell concentration was 4.38×1010 CFU/mL (33℃, 8% milk powder, 1600U/mL bromelain, add 1% glucose at 12 hr, harvest at 24 hr).

In 80L batch fermentation, cell concentration of Lactobacillus acidophilus could reach 2.27×109 CFU/mL (30℃, 10 % milk powder, 1600 U/mL bromelain, add 1 % glucose at 12 hr, stir 10 sec/12hr, harvest at 24 hr); while that of Bifidobacterium bifidum was 1.31×109 CFU/mL (28℃, 8 % milk powder, 1600U/mL bromelain, add 1 % glucose at 12 hr, stir 10 sec/12hr, harvest at 24 hr). For Lactobacillus sporogenes, the cell population was 4.27×109 CFU/mL (33℃, 8 % milk powder, 1600 U/mL bromelain, add 1 % glucose at 12 hr, stir 10 sec/12hr, harvest at 24 hr).

In drying process, the reduced pressure, low temperature fluidized bed spray dryer could produce 8Kg lactic acid bacteria powder, at drying temperature of 33°C, in 36 HR. The cell viability of Lactobacillus acidophilus powder and Bifidobacterium bifidum powder were 108 CFU/g , and that for Lactobacillus sporogenes powder was 109 CFU/g . The loss of cell viability during spraying, caused by pressure drop, was estimated to be 20%.

書名頁
國科會授權書
論文口試委員會審定書(中文版)
論文口試委員會審定書(英文版)
中文摘要 Ⅰ
英文摘要 Ⅲ
誌謝 Ⅴ
目錄 Ⅶ
圖目錄 .ⅩⅡ
表目錄 .ⅩⅤ

壹、 前言 1
貳、 文獻回顧 2
一、 Probiotics（益生菌）and Prebiotics（益菌生） 2
1. 腸道內菌群與健康 2
1-1 腸道內菌群之分佈 2
1-2 腸道內菌群存在之重要性 5
1-3 影響腸內菌相的因子 7
1-4 腸道內菌群對健康之影響 10
2. probiotics (益生菌，原生菌) 11
2-1 益生菌之定義 11
2-2 益生菌應具備之條件 13
2-3 益生菌之作用 19
2-4 益生菌的應用 33
3. Prebiotics（益菌生，益生助生質，益菌益生源） 35
3-1 Prebiotics 之定義 35
3-2 Prebiotics 之條件 35
3-3 Prebiotics 之機能性 35
3-4目前所認定為prebiotics 之物質 36
4. Synbiotics (Conbiotics) 36
二、 乳酸菌(Lactic Acid Bacteria, LAB) 38
1. 乳酸菌的起源 38
2. 乳酸菌之特性 39
3. 乳酸菌在自然界之分布 40
4. 乳酸菌對碳水化合物之代謝 40
5. 環境對乳酸菌基因表現的影響 44
6. 乳酸菌在腸道中的作用機制 47
7. 乳酸菌的生理功能 48
8. 乳酸菌之分類 64
9. 乳酸菌的應用 71
三、主要研究菌株及其優點 75
1. 雙歧桿菌 75
1-1雙歧桿菌之特性 75
1-2 雙歧桿菌在自然界之分布 76
1-3 雙歧桿菌對碳水化合物之代謝 76
1-4 雙歧桿菌對人體的作用 77
2. 孢子型乳酸菌 83
2-1 孢子 83
2-2 孢子型乳酸菌的鑑定特徵 84
2-3 孢子型乳酸菌在腸道的特殊性 84
2-4 孢子型乳酸菌的特點 84
四、低溫負壓式流動床噴霧造粒乾燥機 87
參、 材料與方法 89
一、實驗藥品 89
1. 培養基 89
2. 添加物 89
二、實驗菌株 90
三、實驗方法 90
A. 菌種保存 90
B. 菌株活化 91
C. 液態培養 91
a. MRSB之液態培養及馴養 91
b. 牛乳之液態培養 92
D. 固態培養與菌落測試（Double Layers Plate Count） 92
E . 菌落數之判定 93
F. 80L乳酸菌批次發酵 94
肆、 結果與討論 95
第一章 乳酸菌之篩選與馴養 95
一、 菌種活化 95
二、 篩選與馴養 95
第二章 燒瓶培養 102
一、 培養基 102
1. 主培養基 102
2. 添加物 107
2-1 添加物測定 107
2-2 酵素測定 107
2-3 維生素B 群測定 122
2-4 複方腸胃中藥粉測定 122
二、 pH值 131
三、 溫度 131
四、 最適培養基 144
1. 牛乳濃度 144
2. 酵素濃度 144
3. 醣 144
3-1 Glucose and Lactose 144
3-2 Glucose 濃度 157
五、 最終測定及最高菌體濃度 157
第三章 80L 批次發酵 162
一、 牛乳發酵 162
1. 靜置培養 162
2. 分層測定 162
3. 攪拌測定 162
4. 延長發酵時間 162
二、 羊乳發酵 177
第四章 低溫負壓式流動床噴霧造粒乾燥機之粉劑製造 180
一、 全脂牛奶粉 1850
二、 8Kg 粉底 180
1. 流動劑和乳醣 180
2. 流動劑 183
三、 羊奶粉粉末 186
伍、 結論與展望 189
參考文獻 192
附錄一、Vitamin B complex 成分 216
附錄二、平胃散成分 217
附錄三、實驗室所用之儀器設備 218

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