臺灣博碩士論文加值系統

由兩株菌的混合實驗，發現Aspergillus sp.與Penicillium funiculosum混合培養可得到較強之纖維分解酵素活性，因此以這兩株菌進行混合培養之實驗。而混合培養使用106spores/g DW之總接菌量，Aspergillus sp.與Penicillium funiculosum的混菌比例為1：3，同時接菌培養。培養基的水分含量為80%，木屑以1%NaOH前處理，木屑麩皮比例為1：1，不額外添加氮源，另外添加multiple微量元素，於30°C下培養至第6天，可得到較強之酵素活性，其total cellulase、cellobiohydrolase（CBH）、endoglucanase（EG）與β—glucosidase （BG）之酵素活性分別為2.6985 IU/ g DW、0.465 IU/ g DW、20.6055 IU/ g DW 以及8.4045 IU/ g DW。使用120ml之萃取溶液，而萃取溶液使用0.05M之磷酸緩衝液（pH 6），再添加0.2%之Tween 80，以100 rpm之振盪轉速，在25°C之溫度下萃取一個小時，可以得到最佳之酵素活性，而其total cellulase、EG、CBH與BG之酵素活性分別為4.4476 IU/ g DW、37.8108 IU/ g DW、0.8404 IU/ g DW 以及13.8691 IU/ g DW。且由Asp. sp.與P. f.混合培養所得到的酵素系統，其最適pH為4，最適作用溫度是60°C，pH4~7間酵素活性較穩定，而於50° C下仍能保持80%之活性。而Asp. sp.與P. f.混合培養所得到的酵素系統對無前處理之甘蔗渣即有良好之糖化性，24小時可獲得6%之還原糖，前處理過之甘蔗渣甚至可得到將近20%左右之還原糖。

目錄
中文摘要 I
英文摘要 III
誌謝 V
目錄 VI
圖目錄 X
表目錄 XIII
壹、緒言 1
貳、文獻回顧 3
一、前言 3
二、纖維素之構造 4
三、木質纖維素 7
四、纖維物質的利用轉換性 7
五、纖維分解酵素 10
（一）纖維分解酵素的種類 10
（二）纖維分解酵素的特性 11
（三）纖維分解酵素水解纖維素的作用機制 15
（四）纖維分解酵素之生合成及產物回饋抑制 18
（五）纖維分解酵素之協同作用 23
六、混菌培養（mixed culture）之介紹 24
參、材料與方法 25
一、實驗使用之菌株 25
二、實驗使用之試藥 25
三、實驗設備 28
四、纖維廢棄物之收集與處理 29
五、培養基及各種藥品之配置 30
（一）菌株保存更新用之培養基 30
（二）生產纖維分解酵素用之固態培養基 32
（三）0.2% Tween80 生理食鹽水 32
（四）還原糖測定試劑之配製 32
（五）酵素反應用基質溶液之配製 34
六、實驗方法 35
（一）菌種之保存及使用 35
（二）孢子懸浮液之製作 35
（三）粗酵素萃取 36
（四）纖維分解酵素活性之測定 36
（五）還原糖之測定 37
（六）預備實驗 38
1.所有菌株之預搭配 38
2.木屑以不同方式調濕對纖維分解酵素活性之影響 38
（七）Aspergillus sp.和Penicillium funiculosum之搭配實驗 39
1.不同接菌量對纖維分解酵素活性之影響 39
2.不同混菌比例對纖維分解酵素活性之影響 39
3.不同相化時間對纖維分解酵素活性之影響 39
4.培養基水分含量對纖維分解酵素活性之影響 40
5.不同前處理之木屑對纖維分解酵素活性之影響 40
6.木屑麩皮之比例對纖維分解酵素活性之影響 41
7.添加不同氮源對纖維分解酵素活性之影響 41
8.添加不同之微量元素對纖維分解酵素活性之影響 41
9.培養溫度對纖維分解酵素活性之影響 42
10.最適化條件對纖維分解酵素活性之影響 42
（八）粗酵素萃取方式之探討 43
1.萃取溶液的量對纖維分解酵素活性之影響 43
2.不同萃取溶液對纖維分解酵素活性之影響 43
3.萃取溶液之pH值對纖維分解酵素活性之影響 44
4.不同萃取方式對纖維分解酵素活性之影響 44
5.不同濃度的Tween 80對萃取纖維分解酵素之影響 44
6.萃取溫度對纖維分解酵素活性之影響 45
7.萃取時間對纖維分解酵素活性之影響 45
（九）粗酵素反應特性之探討 45
1.酵素最適作用pH值 45
2.酵素最適作用溫度 46
3.酵素之pH穩定性試驗 46
4.酵素之熱穩定性試驗 46
5.振盪轉速對酵素作用之探討 47
6.木質纖維廢棄物之糖化作用 47
肆、結果與討論 48
一、預備實驗 48
（一）所有菌株之預搭配實驗 48
（二）木屑以不同方式調濕對纖維分解酵素活性之影響 51
二、Aspergillus sp.和Penicillium funiculosum之搭配實驗 53
（一）不同接菌量對纖維分解酵素活性之影響 53
（二）不同混菌比例對纖維分解酵素活性之影響 56
（三）不同相化時間對纖維分解酵素活性之影響 59
（四）培養基中水分含量對纖維分解酵素活性之影響 62
（五）不同前處理之木屑對纖維分解酵素活性之影響 65
（六）木屑麩皮之比例對纖維分解酵素活性之影響 68
（七）添加不同氮源對纖維分解酵素活性之影響 71
（八）添加不同微量元素對纖維分解酵素活性之影響 76
（九）培養溫度對纖維分解酵素活性之影響 80
（十）最適化條件對纖維分解酵素活性之影響 83
三、粗酵素萃取方式之探討 85
（一）萃取溶液的量對纖維分解酵素活性之影響 85
（二）不同萃取溶液對纖維分解酵素活性之影響 87
（三）萃取溶液之pH值對纖維分解酵素活性之影響 89
（四）不同萃取方式對纖維分解酵素活性之影響 91
（五）不同濃度的Tween 80對萃取纖維分解酵素之影響 93
（六）萃取溫度對纖維分解酵素活性之影響 95
（七）萃取時間對纖維分解酵素活性之影響 97
四、粗酵素反應特性之探討 99
（一）酵素最適作用pH值 99
（二）酵素最適作用溫度 101
（三）酵素之pH穩定性試驗 103
（四）酵素之熱穩定性試驗 105
（五）振盪轉速對酵素作用之探討 107
（六）木質纖維廢棄物之糖化作用 109
伍、結論 111
陸、參考文獻 113

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