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標題: 以固定化海藻糖合成酶生產海藻糖之研究
Immobilization of recombinant trehalose synthase for the production of trehalose
作者: 程子鈞
Cheng, Tzu- Chun
關鍵字: trehalose synthase
出版社: 化學工程學系所
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摘要: 海藻糖是一種天然的多功能雙糖。本研究將菌種Picrophilus torridus所生產的海藻糖合成酶,結合純化跟酵素固定化的步驟來生產海藻糖。商業膠體Eupergit C 被選為此研究的固定化基材。經純化後的P. torridus海藻糖合成酶,自由態跟固定化酵素的最適操作pH值均為pH 6,最佳反應溫度皆為50oC,並未隨著酵素作固定化操作而改變。不過,經由固定化操作的酵素展現出較好的溫度耐抗性,在反應溫度為70 oC時,固定化的酵素相較於自由態(4.1 %)能保有較高的殘餘活性59.5 % 。在最大海藻糖轉換率方面從實驗結果得知不論是固定化酵素還是自由態酵素的反應皆與反應基質濃度的高低無關,於最適反應操作50oC、pH 6時,海藻糖最大的轉化率接近60%。對於固定化後批次反應的『可重複使用性』,固定化酵素在重複24個循環操作下依然能保有92.8 %的初始活性。在連續式操作方面,將進料流速控制在0.1ml/min時可得到最大海藻糖的轉化率41.7%。
Trehalose is a kind of natural functional disaccharide. In this study an integrated process following the concomitant purification and immobilization of recombinant trehalose synthase from Picrophilus torridus for the production of trehalose was established. Eupergit C was selected as a carrier in this work. The purified recombinant P. torridus TSase (PTTS) showed that the optimum pH and temperature of 6.0 and 50 °C, respectively, were not alternated upon immobilization. Nevertheless, the immobilized activity did exhibit the higher tolerance of temperature. A higher residual activity of 59.5% was observed for the immobilized enzyme under 70 oC whereas it was 4.1% for free enzyme. The maximum conversion rate of maltose to trehalose by both free and immoblized enzyme was independent upon the substrate concentration under the optimum reaction conditions, in which the yield of approximately 60% for trehalose was obtained. The reusability of the immobilized enzyme in a batch reactor was also studied and we found that the immobilized enzyme maintained approximately 92.8% of residual activity after 24 cycles. While using the continuous packed-bed reactor, a maximum yield of 41.7% for trehalose has been reached from 100 mM maltose with a flow rate of 0.1 ml/min.
其他識別: U0005-1808201111025300
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