臺灣博碩士論文加值系統

本研究以液態培養四株麴菌包括Trichoderma reesei RUT C30， Botrytis cinerea BCRC 33112， Cordyceps militaris ATCC 34164,，Aspergillus niger BCRC 32720以生產纖維素水解酶。以麥麩為基礎培養基，額外補充α-cellulose水解液以及蛋白腖(Peptone)或酵母抽出物(Yeast extract)等氮源。首先培養Asp. niger BCRC 32720，結果發現以麥麩加10%的α-cellulose水解液能使FPU(Filter Paper Assay，纖維水解酵素)，β-glucosdase and CMCase(Endoglucanase，內切纖維水解酵素) 等活性提高28~35 %。α-cellulose水解液扮演誘導劑的角色。
我們用10 %麥麩、0.5 %蛋白腖、1 % α-cellulose水解液來培養四株麴菌以生產纖維素水解酶，結果發現B. cinerea BCRC 33112得到最高的FPU，β-glucosidase，CMCase，分別為0.68 Unit/mL，0.65 Unit/mL，0.58 Unit/mL，而Trichoderma reesei RUT C30 得到最低的活性。我們重複這實驗。我們改用0.5 %酵母抽出物取代蛋白腖，結果也是B. cinerea BCRC 33112得到最高FPU, β-glucosidase, CMCase，分別為0.78 Unit/mL, 0.7 Unit/mL, 0.75 Unit/mL，而Asp. niger BCRC 32720得到第二高的FPU，β-glucosidase, CMCase活性。

The production of cellulose degrading enzymes by cultivation of Trichoderma reesei RUT C30, Botrytis cinerea BCRC 33112, Cordyceps militarisATCC 34164, Aspergillus niger BCRC 32720 was studied. Wheat bran suspension supplemented with α-cellulose hydrolyzate and yeast extract or peptone was used as cultivation medium. It was found that FPU(Filter Paper Assay), β-glucosdase and CMCase activities by Asp. niger 32720 were enhanced in the range of 28~35 % for the medium using wheat bran supplemented with 1 % α-cellulose hydrolyzate. α-Cellulose hydrolyzate performs as an inducer.
Wheat bran (10 %) supplemented with 0.5 % peptone and 1 % α-cellulose hydrolyzate in the medium was used to cultivate the 4 fungi to produce cellulase. The cultivation with B. cinerea BCRC 33112 gave the most activity of FPU (0.68 Unit/ml), β-glucosidase (0.65 Unit/ml), CMCase (0.58 Unit/ml). Trichoderma reesei RUT C30 gave the lowest activity among the four strains.
The experiment was duplicated by replacing 0.5 % peptone to 0.5 % yeast extract. B. cinerea BCRC 33112 again gave the most activity of FPU (0.78 Unit/ml), β-glucosidase (0.7 Unit/ml), CMCase (0.75 Unit/ml). Asp. niger BCRC 32720 ran the second most activity.

誌謝 I
中文摘要 II
Abstract III
圖目錄 VIII
表目錄 X
第一章 緒論 1
1.1研究背景與動機 1
第二章 文獻回顧 2
2.1.1 木質纖維素的組成 2
2.1.2纖維素 3
2.1.4 木質素 6
2.2 纖維素水解酶 7
2.3 纖維素水解酶的生產 8
2.3.1 生產纖維素水解酶的菌種 10
第三章 材料與方法 13
3.1儀器設備與藥品 13
3.2 實驗材料 14
3.2.1菌種來源 14
3.2.2藥品 14
3.3實驗方法 16
3.3.1 菌種保存 16
3.3.2 平板培養 16
3.3.3 液態培養基 16
3.3.4 配置1% α-cellulose水解液 17
3.4 四株麴菌的培養條件 17
3.4.1 Aspergillus niger BCRC 32720在不同濃度水解液 17
3.4.2 四株麴菌額外添加氮源YE 18
3.4.3 四株麴菌額外添加氮源peptone 19
3.5分析方法 20
3.5.1 葡萄糖含量之測定法 20
3.5.2 纖維水解酵素(Cellulase,FPAse)測定(Filter Paper Assay) (Ghose，1987) 20
3.5.3 β-葡萄糖苷酶(β-glucosidase)測定(Tsao et.al,2000) 22
3.5.4 內切纖維水解酵素(Endoglucanase,CMCase)測定 (Ghose，1987) 23
第四章 結果與討論 24
4.1不同濃度α-cellulose水解液的液態培養對纖維水解酵素活性之影響 24
4.1.1 Asp. niger BCRC 32720在不同濃度α-cellulose水解液的FPU 25
4.1.2 Asp. niger BCRC 32720在不同濃度α-cellulose水解液的β-glucosidase 活性 26
4.1.3 Asp. niger BCRC32720在不同濃度α-cellulose水解液的CMCase 27
4.2 比較四株真菌生產纖維水解酶 28
4.2.1 添加0.5 % peptone對纖維素水解酶之影響 29
4.2.2以0.5 %YE 為添加氮源 34
第五章 結論 38
參考文獻 39
附錄 42

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