探討可控式包埋Saccharomyces cerevisiae對於乙醇醱酵之影響

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姓名

黃浩宸(Hao-chen Huang) 查詢紙本館藏

畢業系所

化學工程與材料工程學系

論文名稱

探討可控式包埋Saccharomyces cerevisiae對於乙醇醱酵之影響
(The study of controllable immobilization product ethanol by Saccharomyces cerevisiae)

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摘要(中)

近年來，由於全球石化能源逐漸耗竭及原油價格高漲，尋找永續再生性的新能源已成為一個國家賴以生存及發展經濟所必須努力的目標。厭氧產氫、甲烷，生質乙醇（bioethanol）及生質柴油（biodiesel）等綠色能源的發展受到重視。
本論文以傳統式包埋Saccharomyces cerevisiae 作為控制組，設計立即式包埋法( in-situ immobilization )與可控式包埋法 (controllable immobilization)進行比較，而立即式包埋法可以改善傳統式包埋結構、質傳問題，可控式與立即式的差異在於，多了菌體釋出步驟，使得菌體利用基質的效率更佳。
以100 g/L、200 g/L的葡萄糖，探討立即式與可控式於乙醇醱酵之可行性，結果發現，Saccharomyces cerevisiae 於100 g/L葡萄糖下，立即式、可控式包埋法較傳統式包埋之產率多了約1.35倍，於200 g/L葡萄糖下，在基質與乙醇濃度的抑制下仍有良好的產率，立即式與可控式分別高於傳統包埋1.28與1.15倍，相同條件下，立即式包埋的菌體活性幾乎維持95 %之間，而傳統式會隨之遞減至93 %，導致乙醇總產量與轉化率不及於立即式。

摘要(英)

Ethanol use as a fuel additive or directly as a fuel source has grown in popularity due to governmental regulations and in some cases economic incentives based on environmental concerns as well as a desire to reduce oil dependency.
There are two-type designs which in-situ immobilization and controllable immobilization respectively are to improve the mass transfer problems of traditional immobilization and comparative study of ethanol production by Saccharomyces cerevisiae cells in traditional and in-situ immobilization .
To study the feasibility of ethanol fermentation which in-situ and controllable immobilization can scale up and industrialize , then using 100 and 200 g/L glucose as feedstock in 2.0L bubble column .The results as shown, in 100 g/L glucose , the productivity of in-situ and controllable immobilization more than traditional immobilization by 1.35 times. In 200 g/L glucose, the ethanol concentration and productivity in the matrix is remain stable, and in-situ and controllable immobilization still up to 1.28 and 1.15 times than traditional immobilization. In this condition, the viability of in-situ immobilization almost maintain about 95% , but the viability of traditional immobilization would decrease progressively to 93% .

關鍵字(中)

★ 包埋
★ 乙醇
★ 酵母
★ 可控式

關鍵字(英)

★ immobilization
★ Saccharomyces cerevisiae
★ ethanol

論文目次

摘要 I
Abstract II
誌謝 III
目錄 IV
圖索引 VIII
表索引 XIII
第一章緒論 1
1-1 研究背景 1
1-2 研究目的 1
第二章文獻回顧 2
2-1 真菌 2
2-1-1真菌介紹 2
2-1-2真菌分類表 3
2-1-3酵母菌(Saccharomyces cerevisiae)之介紹 4
2-2 再生能源 9
2-2-1 再生能源之介紹 9
2-2-2 生質能源發展及應用 9
2-2-3 生質乙醇 11
2-3 微生物固定化 17
2-3-1 固定化發展 17
2-3-2 固定化方法 19
2-3-3固定化對生長速率的影響 24
2-3-4固定化對微生物活性的影響 26
2-4海藻酸鈉、海藻酸鈣的介紹 29
2-4-1 海藻酸鈉簡介 29
2-4-2 海藻酸鈉鈣 30
2-4-3 影響海藻酸鈉鈣強度之因子 31
2-4-3-1 海藻酸鈉濃度 31
2-4-3-2 氯化鈣濃度 31
2-4-3-3 浸泡於氯化鈣之時間 31
2-4-3-4 pH值 32
第三章實驗規劃、材料與方法 33
3-1 實驗規劃 33
3-2 實驗材料 33
3-2-1 實驗菌株 33
3-2-2實驗藥品 34
3-2-3 實驗儀器及其他設備 35
3-3實驗方法 36
3-3-1 菌種保存 36
3-3-2 培養基組成 36
3-3-3包埋固定化製備 38
3-3-4酵母菌於不同濃度氯化鈣及海藻酸鈉中的影響 44
3-3-5不同濃度氯化鈣及海藻酸鈉之菌體回收率的研究 45
3-3-6以重複式批次醱酵進行傳統包埋與立即式包埋固定化之研究 45
3-4 分析方法 47
3-4-1 分析流程 47
3-4-2細胞乾重及細胞密度對光學密度之檢量線 48
3-4-3葡萄糖殘量分析： 48
3-4-4乙醇濃度之分析 50
3-4-5 MTS assay 細胞活性測試 51
3-4-6掃描式電子顯微鏡 52
第四章實驗結果與討論 53
4-1比較傳統式包埋、立即式包埋及可控式包埋之製程方法 53
4-2酵母菌於不同濃度的氯化鈣、海藻酸鈉與檸檬酸鈉之影響 54
4-3不同濃度氯化鈣及海藻酸鈉之菌體回收率的研究 58
4-3-1材料特性測試 58
4-3-2立即式與可控式方法之差異 60
4-4搖瓶實驗培養 62
4-4-1重複批次醱酵 (Repeated batch) 62
4-5菌體的表面型態 67
4-6氣泡塔反應槽培養 73
4-6-1傳統式與立即式於100g/L葡萄糖醱酵槽培養之討論 73
4-6-2傳統式與立即式於100g/L葡萄糖醱酵槽之菌體活性測試 73
4-6-3可控式於100 g/L葡萄糖醱酵槽培養之討論 74
4-6-4傳統式與立即式於200 g/L葡萄糖醱酵槽培養之討論 81
4-6-5傳統式與立即式於200 g/L葡萄糖醱酵槽之菌體活性測試 81
4-6-6可控式於200 g/L葡萄糖醱酵槽培養之討論 82
第五章結論與相關建議 89
5-1結論 89
5-2建議 90
第六章參考文獻 91

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指導教授

徐敬衡(Chin-hang Shu)

審核日期

2011-7-18

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