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姓名 王鵬榮(PENG-RONG WANG)  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 探討固態發酵自行生產纖維水解酵素用於分解稻稈及生產生質酒精之研究
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摘要(中) 近年來由於能源危機與石油價格波動,以農業廢棄物作為基質的生質酒精為研究議題漸受重視,也有需多文獻及研究成果,足以說明生質能源目前佔有重要的地位。

本研究透過固態發酵培養Aspergillus niger與Trichiderma reesei,大量生產纖維水解酵素,並於糖化步驟中提出有別於傳統的糖化方法。傳統的方法均為添加純化的纖維水解酵素,近年來亦有自行生產酵素的操作,且需經過粗製的純化與分離菌體的步驟。在本研究中提出藉由控制環境參數以省略分離純化的步驟,抑制菌體生長以及測試還原糖累積,達到減少成本的效果。在糖化的步驟發現纖維水解酵素活性為糖化步驟的主因,進而提出延長酵素活性的方法,使累積還原糖達到更好的效果。最後利用糖化後的發酵液讓Saccharomyces cerevisiae 進行酒精發酵。

由實驗結果顯示,本研究第一階段的Aspergillius niger與Trichoderma reesei共培養酵素液最大活性為0.54FPU/g sub,而在第二階段可以從2g稻稈糖化出的還原糖累積量為14.25g/L,水解效率達到64%,而在第三階段酒精發酵的酒精生產量為3.99g/L,轉化率為55%。

摘要(英) In this study, agricultural waste—straws were hydrolyzed and used to replace the relatively expensive glucose or molasses in ethanol production ,and to reduce the cost of the fermentation process.

Solid-state fermentation was carried out with Trichoderma reesei RUT C-30 and Aspergillus niger BCRC 31130 co-cultured at 28°C. 7.5ml/g sub of 50mM citric acid buffer solution were used to extract enzyme from SSF residual. The crude enzyme without any purification and concentration with the highest enzyme activity of exoglucanase, endoglucanase and β-glucosidase was 0.62, 0.79 and 7.5 U/ml, respectively.

Hydrolysis process was carried out at 50°C for 48h ,with add to soymilk to reduce the cellulase inactivation ,and to get the highest cumulative sugar concentration of 14.25 g/L and the decomposition ratio of 64%.

Ethanol fermentation was carried out in a serum bottle with the sugar gain from hydrolysis and obtained the highest ethanol concentration of 3.99g/L.

關鍵字(中) ★ 生質酒精
★ 發酵
★ 稻稈
關鍵字(英)
論文目次 摘要 II

目錄 IV

表目錄 VI

圖目錄 VII

第一章 緒論 1

1.1. 研究動機 1

1.2. 研究目的 2

第二章 文獻回顧 3

2.1. 木質纖維素 3

2.1.1. 木質纖維素簡介 3

2.1.2. 木質纖維素前處理 5

2.2. 纖維水解酵素 8

2.2.1. 纖維水解酵素簡介 8

2.2.2. 纖維水解酵素來源 9

2.2.3. 纖維水解酵素適合之環境參數 11

2.3. 裡氏木黴菌(Trichoderma reesei) 11

2.4. 黑曲黴(Aspergillus niger) 12

2.5. 蛋白酶抑制劑 13

2.6. 豆漿 14

2.7. 生質酒精 15

2.8. 生質酒精現有技術 16

2.8.1. 分開糖化水解和發酵程序 16

2.8.2. 同步糖化與發酵程序 16

2.9. 釀酒酵母菌(Saccharomyces cerevisiae) 17

第三章 實驗規劃、材料與方法 18

3.1. 實驗材料 18

3.1.1. 實驗菌株 18

3.1.2. 實驗藥品 19

3.1.3. 實驗設備 21

3.2. 實驗流程設計 22

3.2.1. 生產纖維水解酵素 23

3.2.2. 糖化生產還原糖 23

3.2.3. 酒精發酵 24

3.3. 實驗方法 24

3.3.1. 菌種保存 24

3.3.2. 培養基組成 25

3.3.3. 稻稈之前處理 29

3.3.4. 纖維水解酵素生產 30

3.3.5. 糖化生產還原糖 31

3.3.6. 酒精發酵 33

3.4. 分析方法 33

3.4.1. 還原糖定量 33

3.4.2. 酵素活性分析 35

3.4.3. 乙醇濃度分析 37

第四章 實驗結果與討論 39

4.1. 纖維水解酵素生產 39

4.1.1. 固態發酵生產纖維水解酵素 39

4.2. 糖化生產還原糖 43

4.2.1. 酵素萃取比例與還原糖無法持續上升之情形 43

4.2.2. 在高濃度之稻稈糖化結果 45

4.2.3. 探討延長酵素活性之方法 46

4.3. Saccharomyces cerevisiae發酵生產酒精 50

4.3.1. 以不同基質糖化應用於酒精生產 50

第五章 結論與建議 53

5.1. 結論 53

5.2. 建議 54

第六章 參考文獻 55

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指導教授 徐敬衡 審核日期 2015-8-27
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