山羊瘤胃厭氧細菌之纖維水解酵素的特性分析

出版年份: 
2011年
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中文摘要 ................................................1
Abstract..................................................3
致謝.......................................................4
目錄.......................................................5
表目錄.....................................................7
圖目錄.....................................................8
第一章 緒論
1-1前言....................................................9
1-2生質酒精發展............................................10
1-3生質酒精製程............................................11
1-4木質纖維素Lignocellulose............................12
1-4-1纖維素Cellulose......................................13
1-4-2半纖維素Hemicellulose.............................13
1-4-3木質素 Lignin........................................14
1-5纖維素水解酵素......................................14
1-5-1 葡萄糖苷酶...........................................15
1-5-2 內切型纖維素分解酶....................................15
1-5-3 外切型纖維素分解酶....................................15
1-6纖維素分解酵素來源與特性..................................16
1-7研究動機和目的..........................................18
第二章 材料與方法
2-1實驗儀器................................................20
2-2培養基和試劑配方.........................................20
2-3實驗材料與方法..........................................23
2-3-1DNA基本技術操作.......................................23
2-3-2菌株篩選與培養條件.....................................28
2-3-3基因體DNA(Genomic DNA)製備............................30
2-3-4菌株16S rDNA序列鑑定..................................31
2-3-5蛋白纖維素分解酶活性測試................................31
2-3-6SDS-PAGE & CMC Zymogram..............................34
2-3-7基因選殖(Gene cloning) ...............................36
2-3-8重組蛋白大量表現.......................................37
2-3-9重組蛋白的純化........................................40
2-3-10重組蛋白特性分析......................................41
第三章 實驗結果
3-1篩選具纖維素分解能力之菌株...............................44
3-2測試Streptococcus sp.纖維素水解能力.....................44
3-3 Streptococcus sp.之Endo-glucanase蛋白鑑定.............45
3-4 Sg-EGV基因序列分析....................................45
3-5 Sg-EGV蛋白序列分析....................................46
3-6 Sg-EGV表現與純化......................................46
3-7 Sg-EGV重組蛋白生化特性的分析............................47
3-7-1最適反應溫度(optimal temperature)和熱穩定性............47
3-7-2最適反應pH值(optimal temperature)和pH穩定性............48
3-8金屬離子對Sg-EGV活性的影響...............................48
3-9受質特異性..............................................48
第四章 實驗討論............................................50
第五章 結論................................................53
參考文獻..................................................71
附錄......................................................74