本研究選用聚丙烯摻雜馬來酸酐來做為電紡織法的高分子溶液，成功地製備出新穎的固定化擔體材料－PAN-b-MA奈米纖維膜。透過酸酐官能基直接與纖維素分解酵素上的NH2官能基形成共價鍵結，可將酵素固定於奈米纖維膜上。實驗結果觀察到添加馬來酸酐會導致奈米纖維膜上珠狀物的產生，而適量的珠狀物將有助於酵素的活性表現。當聚丙烯與馬來酸酐之重量摻合比例（weight ratio）為1：1時，固定化酵素可以展現出最佳的活性，其值約為450 U/g-material。在最佳的酵素固定化參數下，進行羧甲基纖維素（carboxymethyl cellulose, CMC）水解反應，實驗結果發現，固定化酵素的活性於反應溫度50℃時，水解表現最佳，且不受到反應pH值變化之影響。就酵素穩定性而言，固定化纖維素分解酵素在高溫下的穩定性較游離酵素佳，而在經過6次CMC水解反應後，其相對活性仍可維持在60%左右，與其他材料相較下具有較佳的操作穩定性。總括而言，纖維素分解酵素經固定在PAN-b-MA奈米纖維膜後，進行水解反應時對pH值忍受力高，並且具有較佳的熱穩定性和操作穩定性，充分地顯示出此固定化纖維素分解酵素在工業應用上的潛力。

Electrospinning is a novel process to produce fibrous membranes with nanoscale diameters and properties of high specific surface area as well as porous structure, so electrospun nanofibrous membranes are excellent for filtration application, drug delivery, tissue engineering scaffold and enzyme immobilization.
In this study, nanofibrous membrane, made by poly(acrylonitrile-blending-maleic acid anhydride) (PAN-b-MA), contained anhydride groups for immobilizing cellulase from Aspergillus niger. The average diameter of 150~300 nm was observed from the nanofiber by FESEM and covalent bond formation between enzyme molecule and nanofibrous membrane was confirmed by using FTIR. When the blending ratio of PAN and maleic anhydride ratio was given as 1:1 (w/w), the immobilization efficiency of nanofibrous membrane was superior. The loading efficiency of immobilized enzyme on nanofiber was about 40%, and the specific activity was 450 U/g-material.
The biocatalytic efficiency of immobilized cellulase was examined for carboxymethyl cellulose (CMC) hydrolysis for determining the optimal immobilization conditions in appropriate enzyme concentration of 10 wt%, pH value of 4.6, and reaction time of 90 min at 60℃. Under the optimal immobilization condition, the immobilized enzyme performed well for CMC hydrolysis at reaction temperature of 50℃ but independent of pH value. In addition, the immobilized cellulase demonstrated better thermal stability than free enzyme at higher temperature. Beside, it retained 60% of its initial activity after six repeated batches of reaction.
PAN-b-MA nanofibrous membrane was prepared by electrospinning and successfully used as the support for cellulase immobilization, and it is the potential reusable biocatalyst for hydrolyzing crystalline cellulose.

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