臺灣博碩士論文加值系統

本研究以穀類作為發酵基質，培養Lactobacillus acidophilus ( A菌)、Bifidobacterium bifidum (B菌)、Lactobacillus sporogenes (S菌)，並觀察冷藏後的菌體活性變化。綜合穀粉培養，為A菌效果最顯著；A菌控制組為1.1 E9 CFU/mL；實驗組以綜合穀粉培養後，菌數提高12倍以上，為1.3 E10 CFU/mL（10% w/v綜合穀粉，10% v/v菌液， 0.1% Bromelain，0.1% α-amylase，0.1% β-amylase，1% glucose，33 ℃, 24hr）。經過冷藏4℃，六天的貯存條件下，S菌菌種活性提高16倍，為3.6 E10 CFU/mL，實驗顯示A菌、B菌、S菌冷藏後菌種活性，以S菌表現最顯著。

以脫脂奶粉作為發酵基質，培養A菌、B菌、S菌，觀察冷藏後之活性變化。發酵24小時後，菌數皆可達E9 CFU/mL（8% w/v脫脂奶粉，10% v/v菌液， 0.1% Bromelain，0.1% α-amylase，0.1% β-amylase，1% glucose，A菌、S菌33℃, B菌30℃，24hr）。冷藏4℃，六天的貯存條件下，S菌活菌數提高3.9倍，為8.6 E9 CFU/mL。顯示A菌、B菌、S菌以脫脂奶粉培養時，以S菌冷藏後的活菌數最高。


本研究另外以MRS、燕麥、糙米、黃豆及黑豆作為B菌發酵基質，比較菌種之生長情況。結果以黃豆培養效果最佳，菌數可達2.1 E9 CFU/mL。其次為燕麥培養液發酵，菌數為8.2 E8 CFU/mL。另外，燕麥、糙米、黃豆及黑豆添加脫脂奶粉培養，實驗結果發現，以糙米培養時，菌數增加13倍，為2.6 E9 CFU/mL；推測是其營養成分中，缺少乳酸菌所需要的氮源之緣故。本實驗使用的乳酸菌培養基，包括脫脂奶粉、燕麥、糙米、黃豆及黑豆，成本分析方面顯示，以脫脂奶粉成本最高。使用全穀類發酵，不僅可維持菌體細胞存活率，並節省奶粉之高成本，提供更好的經濟效益與培養基的選擇性。

This research used lactic acid bacteria (LAB) to ferment cereals like soy bean, brown rice and oat for the production of vegetable source lactic acid bacteria powder. The cell viabilities were higher than 108 CFU/mL (Lactobacillus acidophilus and Bifidobacterium bifidum) or 109 CFU/mL (Lactobacillus sporogenes). This study also compared the viabilities of the three strains of LAB. Using mixed cereal powder, L. acidophilus showed best result, cell concentration was 12 times higher than control (mixed cereal powder 10% w/v, seed culture inoculum at 10% v/v, added 0.1% bromelain, 0.1% -amylase, 0.1% -amylase, 1% glucose, L.acidophilus and L. sporogenes 33C, B. bifidum 30C， 24 h). At 4 C storage for 6 days, L. sporogenes was able to grow further to 3.6 E10 CFU/mL (16 times higher than control ) which was most profound among the three strains investigated.
When skimmed milk was used as fermentation broth to culture the three strains of LAB, and observe variations in viabilities after 6 days of 4C storage. After incubation of 24 h, all strains reached cell concentration of about 1.0E9 CFU/mL. After 6 days of 4C storage, L. sporogenes concentration increased to 8.6 E9 CFU/mL (3.9 fold increase), which among the three LABs, was the most significant growth during low temperature storage.

This research also compared the growth of B. bifidum in MRS, oat, brown rice, soybean and black soybean. Among the selected cereals, the strain grew best in soybean milk. The cell concentration reached 2.1 E9 CFU/mL. That for oat was 8.2 E8 CFU/mL. If skimmed milk powder was added, the cell concentration increased to 2.6 E9 CFU/mL. This could be attributed to the additional nitrogen source in the skimmed milk. Economic analysis showed that the cost for skimmed milk was higher than all cereal bases, while cereal fermentation could provide similar, if not higher viability. Therefore, cereal fermentation is an alternative for LAB production that is worth considering in terms of economy.

中 文 摘 要 I
英 文 摘 要 III
致 謝 V
目 錄 VI I
表目錄 X
圖目錄 XI I
第一章、前言 1
第二章、文獻回顧 3
1. 乳酸菌簡介 3
1-1 益生菌 (probiotics) 3
1-1-1 乳酸菌在自然界之分布 3
1-1-2 乳酸菌之特性 4
1-1-3 乳酸菌對碳水化合物之代謝 5
1-2益生菌種應具備之特點 6
1-3 益生菌群於人體之重要性 8
1-4 乳酸菌之分類 9
1-5 乳酸菌之應用 15
1-6 腸道益生菌的代表性菌種 19

2. 全穀類介紹 26
2-2 全穀類定義 26
2-2 全穀類食物 26
2-3 全穀類食物的主要營養成分 27
2-4 全穀類對人體健康益處 28
2-5 燕麥的組成與生理功效 28
2-6 黃豆的組成與生理功效 31
2-7 糙米的組成與生理功效 32
2-8 黑豆的組成與生理功效 33

3. 益生質(prebiotics ) 34
3-1 益生質之定義 34
3-2 益生質( Prebiotics)之條件 35
3-3 益生質之機能性 35
3-4 目前所認定為prebiotics之物質 36

3-4-1抗性澱粉（resistant starch, RS）分類與功能 36
3-4-2膳食纖維(dietary fiber) 39
3-5 共生質(Synbiotics ) 40

第三章、實驗材料與方法 42
3-1 實驗菌株 42
3-2 實驗培養基 42
3-3 實驗添加物 43
3-4 儀器設備 43
3-5 實驗流程 44

第四章、結果與討論 50
4-1 綜合穀粉培養基接種A菌、B菌、S菌及冷藏結果 50
4-2 脫脂奶粉培養基接種A菌、B菌、S菌及冷藏結果 50
4-3 B菌以穀類培養及添加脫脂奶粉後之菌體濃度 51
4-4 培養基成本比較 52

第五章、結論 53
參考文獻 66

表目錄

表 一、應用於發酵食品之主要乳酸菌 18

表 二、Lactobacillus casei and Bifidobacterium lactis菌株在-20C冷凍0-180天期間之菌數變化 38

表4-1、A、B、S菌以綜合穀粉培養及冷藏4℃、6天後之菌體濃度 56


表4-2、A、B、S菌以MRS、脫脂奶粉培養之菌體濃度及冷藏4℃6天後菌體濃度 58

表4-3、B菌以黃豆、燕麥、糙米、黑豆當培養基比較其添加與未添加脫脂奶粉之菌體濃度 62

表4-4、計算脫脂奶粉、燕麥、糙米、黃豆、黑豆、綜合穀粉之成本分析表 65


圖目錄

圖 一、影響益生菌機能性的技術性因素 19
圖 二、Lactobacillus acidophilus型態 20
圖 三、Bifidobacterium型態 22
圖 四、Bifidobacterium bifidum型態 23
圖 五、Lactobacillus sporogenes型態 25
圖 六、燕麥的組成結構圖 29
圖 七、燕麥中的β-葡聚醣結構 30
圖 八、黃豆營養成分之組成比例32
圖 九、糙米結構 33
圖 十、共生質(synbiotic)冰淇淋生產流程39
圖 十一、培養益生菌/益生質的技術演進 41

圖 4-1 A、B、S菌以綜合穀粉培養及冷藏4℃、6天後菌體濃度 57
圖4-2 A、B、S菌以MRS、脫脂奶粉培養及冷藏4℃、6天後之菌體濃度 59
圖4-2.1 A、B、S菌以MRS、脫脂奶粉與綜合榖粉培養之菌體濃度 60
圖4-2.2 比較A、B、S菌以脫脂奶粉與綜合榖粉培養及冷藏4C保存6天後菌體濃度 61

圖4-3 B菌以黃豆、燕麥、糙米、黑豆當培養基比較其添加與未添加脫脂奶粉之菌體濃度 63

圖4-3.1 B菌以10%黃豆培養基在30℃，24小時培養後之平板落菌64

圖4-3.2 B菌以10%燕麥培養基在30℃，24小時培養後之平板落菌64







 

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