本論文主要利用電紡織技術將聚丙烯腈高分子材料製備成奈米纖維膜的形態。使用amidination reaction活化聚丙烯腈上的CN官能基，藉此可與脂肪分解酵素上NH2官能基形成NC-N共價鍵結，達到脂肪分解酵素固定在奈米纖維膜上的目標。在酵素固定化程序研究發現活化時間5分鐘、固定化時間30分鐘、酵素濃度0.5 wt%、固定化溫度50°C及固定化pH值6的條件下，可製備出最佳比活性之固定化酵素。測試酵素安定性方面，固定化酵素儲存於水溶液20天後其活性沒有任何的損失。此外在重複使用次數方面，固定化酵素經10次批次使用後仍保有其原始活性。酵素反應動力學方面，固定化酵素Vmax為18.27 U/mg-protein和Km為88.42 mM與游離酵素的數值差異不明顯。固定化酵素經過安定性及反應動力學分析所展現之特性為相當具有發展潛力，因此進一步將固定化酵素應用於催化大豆油系統反應生成生質柴油。經最適化反應條件研究發現脂肪酸含量50%的油脂10克、甲醇量4克及水量3.84克於30°C下反應24小時即可達到90%的轉化率，且經重複使用10次後其生質柴油轉化率仍維持在81.2%，不但改善了酵素製程之高成本問題，也提升酵素製程生產生質柴油工業化之可行性。

In the study, electrospun polyacrylonitrile (PAN)nanofibrous membrane was activated by amidination reaction for immobilized Pseudomonas cepacia lipase with covalent binding.
For optimized immobilization, different levels of temperature, pH value, immobilization time and enzyme concentration were investigated to determine the optimal specific activity of enzyme. The reaction conditions were obtained as the temperature at 50°C, pH value at 6, immobilization time of 30 min, and enzyme concentration for immobilized lipase of 0.5 wt%. The biocatalytic reaction between enzymatic active site and substrates can be described by using the Michaelis-Menten equation, so the kinetic parameters of immobilized lipase, Km and Vmax, were determined as 88.42 mM and 18.27 U/mg-protein, respectively. Besides, the thermal and pH stabilities of immobilized lipase were improved and still retained the activity as the original one after 10 times repeated uses.
The effects of methanol amount, water content, temperature and content of fatty acid on the conversion of biodiesel were also illustrated and the optimal operation condition was determined. Under the optimal condition, methanol of 4 g, water content of 3.84 g, temperature at 30°C and 10 g oil (contain 50% fatty acid) for biodiesel production, a conversion of 90% was obtained in 24 hours and can remain the conversion of 81.2% after 10 times repeated uses. This effective immobilized lipase demonstrated the high potential for biodiesel production in industrial applications.

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