Trichoderma reesei與Aspergillus niger共醣化稻稈及Saccharomyces cerevisiae生產生質酒精之研究

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研究生：	郭以謙 Yi-chin KUO
論文名稱：	Trichoderma reesei與Aspergillus niger共醣化稻稈及Saccharomyces cerevisiae生產生質酒精之研究 Trichoderma reesei and Aspergillus niger co-saccharification rice straw and production of bioethanol by Saccharomyces cerevisiae
指導教授：	徐敬衡 Ching-heng Shu
口試委員:
學位類別：	碩士 Master
系所名稱：	工學院 - 化學工程與材料工程學系 Department of Chemical & Materials Engineering
論文出版年：	2014
畢業學年度：	102
語文別：	中文
論文頁數：	95
中文關鍵詞：	Trichoderma reesei 、Aspergillus niger 、Saccharomyces cerevisiae 、酵素
相關次數：	點閱：7 下載：0
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摘要
近年來由於能源危機和石油價格高漲，以農業廢棄物做為基質生產生質酒精為研究議題漸受重視，也有許多文獻及研究成果，足以說明生質能源目前占有的地位。
本研究的發酵策略是透過液態單獨培養兩株真菌A.niger及T.reesei分別生產纖維水解酵素，然後以增加接菌量的方式來提升酵素的產量，並且以自行生產的豆渣取代複合式氮源來降低發酵成本，在進入醣化階段時，經過實驗分析醣化之物理條件以及適當的醣化時機後以Mixing的方式混合兩真菌的酵素發酵液進行AT共醣化，在共醣化階段欲發揮AT酵素液水解的極限而在該階段總共添加了兩次稻稈，最後將醣化階段的醣化液應用於Saccharomyces cerevisiae酒精發酵。
由實驗結果顯示，本研究第一階段的AT混合酵素液最大活性為0.511 FPU/mL，而在第二階段可以從3g稻稈醣化出的還原醣累積量為21.25g/L，水解效率達到85.2%，在第三階段酒精發酵的酒精生產量為7.172 g/L，轉化率為76.7%。

Abstract
The study is to develop a fermentation strategies by liquid monocultural two strains of fungi A.niger and T.reesei production hydrolysis enzymes, and then to increase the inoculum way to enhance the production of enzymes, and to replace the nitrogen compound source by self-production of bean dregs to reduce the cost of the fermentation process, when entering the saccharification stage,after study the physical conditions and the timing of saccharification,and then by Mixing a way to mix the enzyme fermentation broth of two fungi called AT co-saccharification.
To achieve maximum effect of hydrolysis of AT enzyme solution,at co-saccharification stage the rice straw was added twice the saccharification stage of the hydrolyzate,used in the final stage of ethanol fermentation by Saccharomyces cerevisiae.Experimental results show that, AT mixed enzyme solution of the present study the maximum cellulase activity was found to be 0.511 FPU / mL, at optimum condition and from the saccharification of 3g rice straw in the second stage were obtained the maximum reducing sugar accumulation of 21.25g / L, the hydrolysis efficiency of 85.2 %, the amount of ethanol produced of 7.172 g / L in the final stage, the conversion yield was 76.7%.

摘要   i
Abstract  ii
誌謝   iii
目錄   v
圖目錄   ix
表目錄  xii
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  4
2-1 木質纖維素  4
2-1-1 木質纖維素簡介   4
2-1-2 木質纖維素之前處理   5
2-2 纖維水解酵素  10
2-2-1 纖維水解酵素的簡介   10
2-2-2 纖維水解酵素來源   10
2-2-3 水解酵素群(Hydrolytic enzyme)  12
2-2-4 纖維水解酵素適合之環境參數  13 
2-3 裡氏木黴菌(Trichoderma reesei)  14
2-4 黑麴黴 (Aspergillus niger)  14
2-5 生質酒精  17
2-6 釀酒酵母菌 (Saccharomyces cerevisiae)   19
第三章 實驗規劃、材料與方法  23
3-1 實驗流程設計  23
3-2 實驗材料  25
3-2-1 實驗菌株 25
3-2-2 實驗藥品 26
3-2-3 實驗設備 28
3-3 實驗方法  30
3-3-1 稻稈之前處理   30
3-3-2 菌種保存 31
3-3-3 培養基組成 32
3-3-4 纖維水解酵素生產  36
3-3-5 醣化生產還原糖   37
3-3-6 Saccharomyces cerevisiae發酵生產酒精   39
3-4 分析方法  40
3-4-1 還原糖定量 40
3-4-2 酵素活性分析   42
3-4-3 乙醇濃度分析   44
3-4-4 菌重定量 45
第四章 實驗結果與討論  46
4-1 生產纖維水解酵素  46 
4-1-1 A niger與T reesei兩菌株種瓶實驗   46 
4-1-2 A niger與T reesei兩菌株酵素活性探討   48
4-1-3 豆渣與複合式氮源活性表現   52 
4-1-4 探討T reesei接菌量及誘導物添加量  53
4-2 醣化生產還原糖  55
4-2-1 醣化物理條件   55
4-2-2 醣化時機的選擇   58 
4-2-3 A niger與T reesei共醣化   63
4-2-4 A niger與T reesei共醣化補充稻稈   65
4-2-5 商業酵素醣化稻稈   69
4-3 酒精發酵  72
第五章 結論與建議  76
5-1 結論  76
5-2 建議  78
第六章 參考文獻  79
                                

第六章參考文獻
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