https://scholars.lib.ntu.edu.tw/handle/123456789/87838
Title: | 利用生物技術大量產製瘤胃細菌纖維分解酵素複合體 | Authors: | 徐濟泰 | Keywords: | 纖維素分解酵素;聚木糖分解酵素;纖維素結合蛋白質;酵素活性;cellulase;xylanase;cellulose;binding protein;enzyme activity | Issue Date: | 2002 | Publisher: | 臺北市:國立臺灣大學動物科學技術學系暨研究所 | Abstract: | 本試驗以瘤胃中之纖維分解菌F.succinogenes S85 為纖維素分解酵素及纖維素結合蛋白質(CBP)來源,並利用R.. albus 7 之XynC 基因產物為聚木糖分解酵素(xylanase)來源,測試添加纖維素結合蛋白質後之酵素活性表現。XynC 基因之產物 約為62 kDa,且具有高活性之聚木糖分解酵素活性。來自F.. succinogenes S85 之纖維素分解酵素經離子交換法及膠體過濾法純化後可得約120 及65 kDa 兩個主要蛋白質,且經活性染色確定其酵素活性。CBP利用含10%纖維二糖之磷酸鹽緩衝液配合1%CHAPS 進行回收。XynC 基因產物之聚木糖分解酵素與純化後之纖維素分解酵素,在加入CBP 後可有效提升其酵素作用 之特異性4 至10 倍,且發現CBP 可作用於不同菌株而來之分解酵素,提升酵素之效力。 Attempts were made to examine enzymes activity after addition cellulose binding protein (CBP). Cellulase and CBP were purified from rumen cellulytic bacterium F.. succinogenes S85, and the gene encoding xylanase C (XynC) of rumen cellulytic bacterium R. albus 7 was be the source of xylanase. The gene encoded xylanase was 62 kDa, and detected high xylanase activity. Cellulase from F.. succinogenes S85, after passing ion exchange and gel filtration columns, present two protein peaks with cellulase activity, and the molecular weight was approximately 120 and 65 kDa, respectively. CBP was eluted with sodium phosphate buffer containing 10% cellobiose and 1% CHAPS. Enzymes activity were increasing from four to ten times after addition of CBP. The result showed that CBP can effect on both cellulytic and hemicellulytic enzyme from different bacteria species and improve their enzyme activity. |
URI: | http://ntur.lib.ntu.edu.tw//handle/246246/16449 | Other Identifiers: | 902313B002313 | Rights: | 國立臺灣大學動物科學技術學系暨研究所 |
Appears in Collections: | 動物科學技術學系 |
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902313B002313.pdf | 197.85 kB | Adobe PDF | View/Open |
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