由於脂肪分解酵素酵素催化油脂水解生產脂肪酸之製程，具有反應條件溫和及副產物少的優點，優於用化學觸媒催化反應之製程，且酵素具良好生物可分解性，對環境較友善，然酵素價格昂貴，增加製程生產成本。為降低酵素製程的生產成本並提高其發展潛力，因此發展固定化酵素技術，提高酵素穩定性與重覆使用性。由於氧化鐵（Fe3O4）磁性奈米粒子具高化學穩定性與生物相容性，且具備磁性易回收，近年來被廣泛作為酵素固定化之載體。
　　在本研究中，將脂肪分解酵素（lipase）固定化在氧化鐵（Fe3O4）磁性奈米粒子上，並應用於大豆油脂水解生成脂肪酸之研究。脂肪分解酵素經碳二醯胺（carbodiimide）活化後，可鍵結於氧化鐵磁性奈米粒子上。由X-ray繞射光譜（XRD）、穿透式電子顯微鏡（TEM）與超導量干涉磁量儀（SQUID）之物性分析結果，顯示固定化前後磁性奈米粒子結構不變，為奈米級程度，且具有超順磁性。由傅立葉轉換紅外線光譜儀（FTIR）的分析，脂肪分解酵素以共價鍵結在磁性奈米粒子上。在奈米粒子添加濃度為40 mg/mL、固定化pH值為7、固定化時間為60分鐘及固定化溫度為30℃的酵素固定化條件下，固定化酵素具有較高的蛋白質固定量與最佳水解活性。將製備之固定化酵素應用於大豆油水解反應之研究，利用實驗設計法探討反應溫度、油水莫耳比與固定化酵素量對於油脂水解反應之影響，並建立動力學模型描述此反應之行為。由實際實驗值與模型計算值的相關係數（R2）為0.97，說明此模型的可信度。進一步以此模型預測油脂水解反應之最適化反應條件，在此條件下反應23小時左右，脂肪酸產率可達99%，實驗結果與模型預測預測結果相當一致。在酵素之重複使用性實驗部份，本研究所製備的固定化酵素，在重覆使用5次後，其脂肪酸產率為第一次使用之43 %。

The performance of enzyme in catalyzing the oil hydrolysis reaction is better than chemical catalyst, because the enzymatic reaction can operate under a mild reaction condition, produce few by-products, and be friendly to environment. The main purpose of this thesis is to reduce the cost of enzymatic process and to increase the stability and reusability of enzyme. Recently, the technique of immobilization enzyme has been investigated and Fe3O4 magnetic nanoparticles has a novel matrix of immobilization because it has chemical stability and biocompatibility.
In this study, the Candida rugosa lipase was immobilized onto magnetic nanoparticles by carbodiimide activation and employed to catalytically produce fatty acids by hydrolysis of soybean oil. The physical property of magnetic nanoparticles was analyzed by transmission electron microscopy (TEM), X-ray diffraction (XRD) and magnetic measurement (SQUID). It showed that the size and structure of magnetic nanoparticles did not significantly change and possessed superparamagnetic characteristic after enzyme immobilization. Lipase bound onto the magnetic nanoparticles was confirmed by the Fourier transform infrared (FTIR) spectroscopy measurement. The operation conditions for lipase immobilization were optimized to obtain the highest protein loading and specific activity. Hence the immobilized lipase was further employed to investigate the kinetics of the hydrolysis of soybean oil. The hydrolysis reaction was studied at the different levels of reaction temperature, molar ratio of oil to water and lipase amount . Therefore, a kinetic model coupling with the mentioned reaction parameters were proposed to describe the enzymatic hydrolysis of soybean oil. Since the kinetic model met the experiments well, it was applicable to determine the optimal operating condition. Under the optimal reaction conditions, the FFA yield could reach 99% after reaction for 23 hours. An experimental verification was carried out and well matched the model prediction. In reusability examination, it still retained 43% of its initial yield after being used 5 times.

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