Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/3645
標題: 以固定化酵素生產L-同苯丙胺酸
Enzymatic process for the synthesis of L-Homophenylalanine
作者: 顏敏智
Yen, Min-Chih
關鍵字: L-homophenylalanine
L-同苯丙胺酸
L-N-Carbamoylase
racemase
enzyme immobilization
酵素固定化
出版社: 化學工程學系所
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摘要: 本研究主要目的是利用固定化酵素生產抗高血藥物之前驅物L-同苯丙胺酸(L-HPA),這些含有L-HPA的ACEI (Angiotensin-converting enzyme inhibitor )引起的副作用明顯降低了許多,因此,如能以酵素方法大量製造是極具實用性的;固定化擔體選用含有環氧基的高分子聚合物Eupergit C,利用共價鍵結法固定酵素LNCA與Racemase,並探討最適固定化反應條件與固定化酵素之性質,當LNCA 濃度1.05 mg/ml、Racemase濃度0.97 mg/ml為最適固定化酵素之濃度;固定化LNCA最適反應溫度與自由態相比可提高至50℃且溫度操作範圍較大,而固定化Racemase最適反應溫度為60℃,與自由態並無差異;熱穩定性方面,固定化LNCA與Racemase自60~80℃皆比自由態酵素有較高之相對活性;至於反應pH值方面,固定化LNCA與自由態相比在pH 5~7有較高之相對活性,固定化Racemase最適反應pH值為6-9與自由態相比(pH 6~7)相比,往鹼性方向增加兩個單位。 於重複批次反應上,固定化LNCA可維持9個循環,操作時間達18小時,依然保有約70%的初始活性,而固定化Racemase穩定性更佳,經過40個循環,長達80小時的操作時間,活性保有約75%的初始活性;為了找出最適添加比例,本研究利用酵素反應器,以連續進料(1ml/hr)方式得到固定化酵素最適添加比例為1:10 ( Racemase : LNCA );利用酵素固定化方法生產L-同苯丙胺酸,除了對環境的耐抗性有所提昇外,重複使用也是一大優點,可減少酵素使用量,更符合經濟效益。
An enzymatic process was developed for the production of L-homophenylalanine(L-HPA), an angiotensin-converting enzyme inhibitor precursor for the synthesis of many antihypertensive drugs. Eupergit C containing epoxy groups was selected as the support for covalent immobilization of L-N-carbamoylase(LNCA) and racemase. The optimal enzyme loads of LNCA and racemase were 1.05 mg/ml and 0.97 mg/ml, respectively. Compared with the free enzyme, the optimal reaction temperature of immobilized LNCA was increased form 40℃ to 50℃. However, the optimal reaction temperature of racemase was not affected by immobilization. While the optimal pH of LNCA was downshifted by 3 units upon immobilization, that of racemase was broadened upward by 2 pH units. In batch operation, racemase was much more stable than LNCA. While the immobilized LNCA retained 70% of the initial activity after nine cycles, the immobilized racemase retained 75% of the initial activity after forty cycles. The optimal enzyme ratio of racemase and LNCA was 1 to 10 in continuous process. It was shown feasible to produce L-HPA via the novel enzymatic process.
URI: http://hdl.handle.net/11455/3645
其他識別: U0005-2508200717021600
文章連結: http://www.airitilibrary.com/Publication/alDetailedMesh1?DocID=U0005-2508200717021600
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