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標題: 以聚麩胺酸為擔體探討脂解酵素AY共價固定化及其應用在連續式填充床生物反應器之研究
Covalent Immobilization of Lipase AY onto Poly(Gamma-Glutamic Acid) and Its Application in Continuous Packed-Bed Bioreactor
作者: 謝淳仁
關鍵字: 應用研究;Lipase AY;食品科技;Lipase AY;固定化;聚麩胺酸;填充床生物反應器;最優化;Immobilization;Poly(γ-glutamic acid);Packed-bed bioreactor,Optimization
脂解酵素AY 由Candida rugosa 發酵所產生,是最具有發展潛力的商業脂解酵素之ㄧ。它為無特定位專一性的脂解酵素,能夠進行水解和酯化等反應,可廣泛應用在各種工業用途上。脂解酵素AY 為一種水溶性分子,有無法回收重複利用之缺憾,增加了工業製程上成本負擔。因此,脂解酵素AY 固定化之目的是希望增加酵素穩定性,避免於因環境的影響而變性,並提升其特異性、應用範圍及降低成本,以符合經濟利益。而固定化方法的選擇依據酵素、擔體的特性及固定化後的用途來決定,因此許多擔體種類和酵素固定化方法因應而生。本計畫中,聚麩胺酸(Poly γ-glutamic acid)是一種便宜且具有生物降解特性的擔體材料,它由微生物發酵生產的生物高分子,常被應用於藥物的載體、生醫材料及高吸水性水膠領域上。本計畫預計於三年內完成。第一年,分別以吸附法和共價鍵結法將商業脂解酵素AY 和基因改良之C. rugosa 脂解酵素固定於矽藻土(CeliteR 545 coarse)和聚麩胺酸上,進行四種固定化實驗條件之探討。第二年,依第一年所得之固定化條件,利用部份因子實驗設計、統計分析及等高線圖,探討最優化固定化條件。第三年,模擬工業化生產流程,以固定化酵素配合填充床(Packed-bed)生物反應器合成己醇酯類。

Lipase AY from Candida rugosa is one of the most attractively commercial lipases.It has also been widely used for the complete hydrolysis of triacylglycerol and severalesterifications in organic medium. At present, the main disadvantage for the enzymeuse for industrial production is the cost of enzyme. The immobilization of lipase is avalid approach, because it would allow the reutilization and additional stability.Therefore, immobilized lipases in the bioreactors are less expensive, more stable andreusable alternatives to free lipases. The choice of immobilization depends on thesource of lipase, the type of support and the purpose of process. In this project,poly( γ -glutamic acid), γ -PGA, is inexpensive and biodegradable as supportmaterial. γ-PGA, produced by microbial fermentation, offers a wide range of uniqueapplications including being used as drug carriers, biomedical materials, and highlywater absorbable hydrogels.This project plans to be finished in three years. In the first year, commercial lipaseAY and genetic engineered C. rugosa lipase were immobilized onto CeliteR 545coarse by adsorption and γ-PGA by covalent binding, respectively. In the secondyear, the optimum immobilization conditions will be obtained by experimental designand statistical analysis. In the third year, the immobilization lipases will be employed tocatalyze the direct-esterification of hexyl esters in a packed-bed bioreactor.
其他識別: NSC95-2313-B005-085-MY3
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