臺灣博碩士論文加值系統

在眾多澱粉質生質能源作物中，適合台灣地理氣候、容易種植栽培、生長期短且產量高的甘藷被評估為具潛力的能源作物料源。然而甘藷除了利用地下部的澱粉塊根外，佔有 50% 生質料之地上部亦值得注目加以開發利用。本研究以目前台灣產量最高的台農57號甘藷之莖蔓枝葉採收廢棄物為原料，成份分析顯示廢棄物中約含有 64% 的碳水化合物和粗纖維具有被利用來生成生質酒精之潛力。以球磨機進行微米化或利用快速釋壓操作將甘藷農作廢棄物進行前處理後再以 Trichoderma reesei ATCC 56765 所生產的酵素水解糖化；最後以 Sacharomyces cerevisiae BCRC 21679 發酵生產酒精。利用 Trichoderma reesei 與 Aspergillus niger 生產的酵素一起搭配使用可將葡萄糖產量由15.7g/L 增加至 20.0g/L。以 Sacharomyces cerevisiae BCRC 21679發酵150 g/L 經快速釋壓前處理過的甘藷廢棄物產生的糖化液 (24.4 g/L 葡萄糖) 可生產 0.96% 的生質酒精；發酵150 g/L 經球磨機微米化前處理的廢棄物基質產生的糖化液 (22.2 g/L 葡萄糖) 可生產 0.92% 的生質酒精。由於酵素糖化生產的可利用糖產率不高，即使酒精理論產率高，最終酒精濃度也無法顯著提昇。建議未來著重於改善前處理和最適化酵素水解條件以及開發五碳糖糖化與發酵製程以增加廢棄物的利用效果。甘藷纖維素生質酒精的技術開發可減少生質物料浪費有助於降低整體成本，增加生質酒精在生產成本上的競爭力。

Among lots choices of energy crops, sweet potato is considered most suitable for Taiwan. However, currently only the underground starch stem is used for producing bioehtanol, there still 50% sweet potato biomass is not exploited yet. The agricultural waste from currently Taiwan maximum yield sweet potato strain Tainung No. 57 was studied for better bioethanol production. Composition analytical results show the potential that the raw dry material contains approximately 64% carbohydrate and fiber can be used as bioethanol substrate. In this study, sweet potato field waste was used as substrate and processed with quickly pressure release process and ball milling micron treatment. Pretreated substartes saccharified by Trichoderma reesei ATCC 56765 were subsequently fermented by Sacharomyces cerevisiae BCRC 21679 to produce bioehtanol. The saccharified enzymes combination from Trichoderma reesei ATCC 56765 and Aspergillus niger BCRC 21679 improve the glucose yield from 15.7g/L to 20.0g/L. 0.96% enthanol can be acheived from 150 g/L quickly pressure release pretreated substrate through the saccharification (24.4 g/L glucose) and fermentation process. 0.92% enthanol can be harvested from 150 g/L ball milling pretreated micron substrate through the saccharification (22.2 g/L glucose) and fermentation process. However, only parts of the carbohydrate and fiber were utilized in this process. Even though the theoretical yield of ethanol is high, the small amount of fermentable sugar after saccharification stage limited final ethanol yield. Further study on optimized the pretreatment processes, enzyme saccharification and development of pentose saccharification and fermentation should be emphasized to improve the waste utilization.

致謝辭……………………………………………………………………I
中文摘要……………………………………………………………… II
SUMMARY……………………………………………………………… III總目錄………………………………………………………………… IV
表目………………………………………………………………… VII
圖目錄………………………………………………………………IX
第一章 前言…………………………………………………………1
1.1 生質能源的發展 1
1.2 生質酒精 2
1.2.1 生質酒精的歷史及發展現況 2
1.2.2生質酒精的優勢 4
1.2.3 生質酒精原料 7
1.3 應用廢棄物開發生質能源 7
1.4 國內農作廢棄物之處理現況 8
1.5 植物細胞壁組成結構與其應用 9
1.5.1 全纖維素 10
1.5.2 纖維素 10
1.5.3 半纖維素 10
1.5.4 木質素 10
1.6 纖維素糖化技術 11
1.6.1 利用前處理方法幫助糖化水解 11
1.6.2 利用酵素水解農作廢棄物中之纖維素 14
1.7 酒精發酵 15
1.8 研究動機與目的 15
第二章 材料與方法……………………………………………… 16
2.1 實驗材料 16
2.1.1 甘藷農作廢棄物 16
2.1.2 試驗菌株及酵素 16
2.1.3 化學藥品 16
2.1.4 儀器設備 17
2.1.5 培養基 19
2.2 實驗設計與流程 21
2.3 材料組成分分析項目 24
2.3.1 甘藷農作廢棄物組成分測定方法 24
2.4 前處理的條件及分析 28
2.4.1 快速釋壓前處理 28
2.4.2. 使用球磨機進行微米化前處理 29
2.4.3 前處理效果評估 29
2.5 酵素糖化法及分析 31
2.5.1 菌種活化及繼代培養 31
2.5.2 菌種保存 31
2.5.3 斜面培養與孢子懸浮液振盪培養 31
2.5.4 纖維素酵素之活性測定 32
2.5.4.1 CMCase 活性測定 34
2.5.4.2 β-glucosidase 活性測定 38
2.5.5 甘藷農作廢棄物粉末之酵素糖化條件 40
2.6 酒精發酵及其分析 42
2.6.1 不同基質濃度糖化後以S. cerevisiae BCRC21679 進行酒精發酵 44
2.6.2 發酵條件探討 44
2.6.2.1 發酵前滅菌操作對酒精產量之影響 44
2.7 統計分析 44
第三章 結果與討論……………………………………………… 45
3.1甘藷農作廢棄物之組成份分析 45
3.2未前處理甘藷農作廢棄物水解糖化效果 46
3.2 前處理操作 49
3.2.1 快速釋壓前處理之效果評估 49
3.2.1.1 還原糖產量 51
3.2.2 以球磨機進行前處理之效果評估 53
3.2.2.1 利用球磨機前處理造成樣品粒徑改變 53
3.2.2.2 評估經球磨機前處理後對水解產生還原糖產量的影響 55
3.3 添加其他酵素以幫助纖維素水解 57
3.3.1 以培養微生物生產纖維水解酵素糖化之條件最適化 57
3.3.1.1培養 Trichoderma reesei ATCC 56765 以生產纖維水解酵素添加於甘藷廢棄物基質中糖化反應之溫度最適化 57
3.3.1.2添加 Trichoderma reesei ATCC 56765 以生產纖維水解酵素於甘藷廢棄物基質中進行糖化反應之酵素添加量探討 60
3.3.2 供反應甘藷廢棄物基質濃度最適化 62
3.3.2.1 以來自 Trichoderma reesei ATCC 26921 生產的商業酵素處理不同基質濃度之甘藷廢棄物 62
3.3.2.2 以的Trichoderma reesei ATCC 56765 所生產的纖維水解酵素處理不同濃度的甘藷廢棄物 66
3.3.3 混合兩種菌種生產水解酵素糖化甘藷廢棄物之最適化 70
3.3.4 前處理對酵素水解之影響 74
3.5 酒精發酵 78
3.5.1不同甘藷廢棄物基質濃度經不同前處理操作後糖化以S. cerevisiae BCRC21679進行酒精發酵 78
3.5.1.1培養 Trichoderma reesei ATCC 56765 生產水解酵素添加於未經過前處理之甘藷廢棄物基質中進行糖化反應 78
3.5.2 培養的發酵條件探討 80
3.5.2.1 糖化後滅菌操作對酒精產量之影響 80
3.5.3 結合前處理、水解條件與發酵條件 89
第四章 結論與展望……………………………………………… 94
參考文獻 ………………………………………………………………96

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