臺灣博碩士論文加值系統

纖維分解酵素是可分解纖維素的水解酵素(hydrolyticenzyme)的總稱，經由內切、外切、β-葡萄糖分解酵素等彼此間的作用，將纖維素分解成可溶性的葡萄糖。本實驗分別測試芽孢桿菌Bacillus amyloliquefaciens BA(CY)與B. amyloliquefaciens A1的CMCase、Avicelase及β-glucosidase在不同溫度下的酵素活性和還原糖量。從剛果紅染色結果中發現兩株菌呈現明顯的透明環，可知兩株菌皆具有纖維降解能力。由內切、外切及β-葡萄糖分解酵素試驗得知，兩株菌在溫度60℃時有最適合之酵素活性，且在BA(CY)與A1的菌液與上清液比較中發現菌液分解出的葡萄糖量較上清液高。從稻稈纖維降解試驗得知，A1在前12小時其分解出的還原糖比BA(CY)高，但A1至48小時之後從391(µg/mL)降至292(µg/mL)，BA(CY)在48小時其還原糖量從274(µg/mL)上升至310(µg/mL)，活性也相對降低。由豆渣纖維降解試驗得知，BA(CY)與A1皆持續上升，於132小時時BA(CY)還原糖量為629(µg/mL)，A1為466(µg/mL)。

Cellulase are the general term for a group of hydrolytic enzymes, which break down cellulose into soluble glucose through synergism between endo, exo, and β-glucose decomposing enzymes. This study tests the enzyme activity and reducing sugar amount of CMCase, Avicelase and β-glucosidase of Bacillus amyloliquefaciens BA(CY) and B.amyloliquefaciens A1 at different temperatures. From the results of Congored staining, it was found that the two strains showed obvious transparent rings, which indicated that both strains had fiber degradation ability. It is known from endo, exo and β-glucose decomposing enzyme tests that the two strains had the most suitable enzyme activity at 60℃, and the bacteria were found in the comparison of the bacterial liquid and supernatant of BA(CY) and A1 the amount of glucose decomposed from the liquid is higher than that of the supernatant. The rice straw fiber degradation test, it is known that A1 decomposes higher reducing sugars than BA(CY) in the first 12 hours, but A1 drops from 391(μg/mL) to 292(μg/mL) after 48 hours. The amount of reducing sugar increased from 274(µg/mL) to 310(µg/mL), and the activity was relatively reduced. According to the okara fiber degradation, BA(CY) and A1 continued rise. At 132 hours, the amount of reducing sugar in BA (CY) was 629(µg/mL), andA1 was 466(µg/mL).

目錄
圖目錄 xi
表目錄 xiv
壹、前言 1
一、循環經濟 CircularEconomy 1
二、纖維分解酵素 Cellulase 4
三、Bacillus spp. 6
四、研究目的 7
貳、材料與步驟   9
一、菌種的來源、純化和保存 9
(一) 菌種來源 9
(二) 菌種的純化 9
(三) 菌種的保存 9
二、纖維降解試驗 10
(一) 固態纖維分解酵素活性測試 10
(二) 分解纖維酵素活性分析-DNS還原糖試驗 11
三、BA(CY)與A1纖維降解試驗分析 13
(一) BA(CY)、A1上清液在60℃、24小時下內切、外切及β-葡萄糖還原糖與酵素活性分析 13
(二) BA(CY)、A1菌液在不同溫度下內切、外切及β-葡萄糖之還原糖與酵素活性分析 14
(三) BA(CY)、A1上清液與菌液在60℃、24小時之還原糖與酵素活性分析 14
四、稻稈纖維降解試驗 15
五、豆渣纖維降解試驗 15
參、結果 16
一、纖維降解試驗結果 16
(一) 固態纖維分解酵素活性測試結果 16
(二) 分解纖維酵素活性分析-DNS還原糖試驗結果 16
二、 BA(CY)與A1纖維降解試驗分析結果 17
(一) BA(CY)、A1上清液在60℃、24小時內切、外切及β-葡萄糖還原糖與酵素活性分析 17
(二) BA(CY)、A1菌液在不同溫度下內切、外切及β-葡萄糖之還原糖與酵素活性分析結果 18
(三)BA(CY)、A1菌液在60℃、24小時內切、外切及β-葡萄糖還原糖與酵素活性分析 18
三、稻稈纖維降解試驗分析結果 20
四、豆渣纖維降解試驗分析解果 20
肆、討論 21
伍、參考文獻 24
陸、圖表 28
柒、附錄 67圖目錄

圖一、於甲基纖維素生長培養基以剛果紅染色之結果 28
圖二、於甲基纖維素生長培養基以剛果紅染色之結果 29
圖三、葡萄糖標準曲線 30
圖四、內切型分解酵素之還原糖量 31
圖五、內切型纖維素分解酵素活性分析結果 32
圖六、外切型分解酵素之還原糖量 33
圖七、外切型纖維素分解酵素活性分析結果 34
圖八、β-葡萄糖分解酵素之還原糖量 35
圖九、β-葡萄糖酵素活性分析結果 36
圖十、BA(CY)、A1上清液在60℃、24小時下之內切型分解酵素(Endo-β-1,4-glucanaese)還原糖量 37
圖十一、BA(CY)、A1上清液在60℃、24小時下之外切型分解酵素(Exo-β-1,4-glucanse)還原糖量 38
圖十二、BA(CY)、A1上清液在60℃、24小時下之葡萄糖分解酵素(β-1,4-glucosidase)還原糖量 39
圖十三、BA(CY)、A1上清液在60℃、24小時下之內切型分解酵素(Endo-β-1,4-glucanaese)活性分析 40
圖十四、BA(CY)、A1上清液在60℃、24小時下之外切型分解酵素(Exo-β-1,4-glucanse)活性分析 41
圖十五、BA(CY)、A1上清液在60℃、24小時下之葡萄糖分解酵素(β-1,4-glucosidase)活性分析 42
圖十六、BA(CY)、A1上清液(A)、菌液(B)在不同溫度下之內切型分解酵素(Endo-β-1,4-glucanaese)還原糖量 43
圖十七、BA(CY)、A1上清液(A)、菌液(B)在不同溫度下之外切型分解酵素(Exo-β-1,4-glucanse)還原糖量 44
圖十八、BA(CY)、A1上清液(A)、菌液(B)在不同溫度下之葡萄糖分解酵素(β-1,4-glucosidase)還原糖量 45
圖十九、BA(CY)、A1上清液(A)、菌液(B)在不同溫度下之內切型分解酵素(Endo-β-1,4-glucanaese)活性分析 46
圖二十、BA(CY)、A1上清液(A)、菌液(B)在不同溫度下之外切型分解酵素(Exo-β-1,4-glucanse)活性分析 47
圖二十一、BA(CY)、A1上清液(A)、菌液(B)在不同溫度下之葡萄糖分解酵素(β-1,4-glucosidase)活性分析 48
圖二十二、BA(CY)、A1上清液(A)、菌液(B)在24小時下之內切型分解酵素(Endo-β-1,4-glucanaese)還原糖量 49
圖二十三、BA(CY)、A1上清液(A)、菌液(B)在24小時下之外切型分解酵素(Exo-β-1,4-glucanse)還原糖量 50
圖二十四、BA(CY)、A1上清液(A)、菌液(B)在24小時下之葡萄糖分解酵素(β-1,4-glucosidase)還原糖量 51
圖二十五、BA(CY)、A1上清液(A)、菌液(B)在24小時下之內切型分解酵素(Endo-β-1,4-glucanaese)酵素活性 52
圖二十六、BA(CY)、A1上清液(A)、菌液(B)在24小時下之外切型分解酵素(Exo-β-1,4-glucanse)酵素活性 53
圖二十七、BA(CY)、A1上清液(A)、菌液(B)在24小時下之葡萄糖分解酵素(β-1,4-glucosidase)酵素活性 54
圖二十八、稻稈在156小時之還原糖分析 55
圖二十九、稻稈0至48小時分解變化圖 56
圖三十、稻稈72至120小時分解變化圖 57
圖三十一、豆渣在132小時之還原糖分析 58
圖三十二、豆渣0至48小時分解變化圖 59
圖三十三、豆渣72至120小時分解變化圖 60
表目錄

表一、內切型分解酵素之還原糖產量 61 08D0C9EA79F9BACE118C8200AA004BA90B02000000080000000D0000005F0054006F006300350033003800310039003400330030000000
表二、內切型纖維素分解酵素活性分析結果  PAGEREF _Toc53819432 \h 62 08D0C9EA79F9BACE118C8200AA004BA90B02000000080000000D0000005F0054006F006300350033003800310039003400330032000000
表三、外切型分解酵素(Avicelaes)之還原糖產量 PAGEREF _Toc53819433 \h 63 08D0C9EA79F9BACE118C8200AA004BA90B02000000080000000D0000005F0054006F006300350033003800310039003400330033000000
表四、外切型纖維素分解酵素活性分析結果  PAGEREF _Toc53819434 \h 64 08D0C9EA79F9BACE118C8200AA004BA90B02000000080000000D0000005F0054006F006300350033003800310039003400330034000000
表五、β-葡萄糖酵素(β-1,4-glucosidase)之還原糖產量  PAGEREF _Toc53819435 \h 65 08D0C9EA79F9BACE118C8200AA004BA90B02000000080000000D0000005F0054006F006300350033003800310039003400330035000000
表六、β-葡萄糖酵素活性分析結果 66 08D0C9EA79F9BACE118C8200AA004BA90B02000000080000000D0000005F0054006F006300350033003800310039003400330036000000

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