由聚丙烯腈包覆之氧化鐵（Fe3O4）磁性奈米粒子，可透過amidination reaction活化粒子表面之聚丙烯腈的C≡N官能基,與纖維素分解酵素上之胺基形成共價鍵結。於實驗條件為活化時間2.5分鐘，固定化酵素濃度為0.5wt%，固定化時間60分鐘，固定化溫度50℃,pH值為7.8的條件下，可製得較高蛋白質固定量之固定化纖維素分解酵素。本實驗進而將固定化酵素應用於水解微藻細胞壁，並藉由產生還原醣之濃度，決定最佳反應條件。經實驗證實在適當的水解時間下，固定化纖維素分解酵素可水解微藻之細胞壁，使藻體細胞壁破裂，但破裂程度不致使葉綠素釋出胞外，造成損失，並可提高後續以丙酮萃取葉綠素之效益。經固定化纖維素分解酵素水解後之微藻，利用丙酮於常溫下進行萃取反應，100 mg之藻體可萃得葉綠素1.5mg；相較未經過水解反應之藻體，則只可萃得0.4mg之葉綠素。重複進行批次反應10次以後，萃取率仍維持初始萃取率之88%。由於氧化鐵磁性奈米粒子具備磁性易回收，因此實驗所製得之固定化纖維素分解酵素，證明不僅可多次重複使用，並具有回收容易之優點。

The enzyme, Trichoderma reesei cellulase was immobilized on PAN (polyacrylonitrile) coated magnetic nanoparticles(Fe3O4) by amidination reaction for long term operation. In addition, covalent bond formation between enzyme molecule and the PAN coated magnetic nanoparticles was confirmed via FT-IR measurement. The optimal operation conditions of cellulase immobilization for the highest protein loading were under the enzyme concentration of 0.5 wt%, immobilization time of 60 min, temperature at 50°C and pH value at 7.8. The optimal conditions of hydrolysis reaction were determined by measuring the concentration of producing reducing sugar. The immobilized cellulase was further employed for hydrolyzing the cell walls of microalgae. The experimental results indicate that under the specific reaction time, cell walls of microalgae were broken without releasing of chlorophyll. After breaking the cell walls of microalgae by hydrolysis reaction with immobilized cellulase, 1.5 mg chlorophyll was extracted with acetone, on the other hand, only 0.4 mg chlorophyll was obtained from the microalgae without hydrolysis before extraction. In reusability experiments, the efficiency retained 88% of its initial value after 10 time uses of the immobilized cellulase. Since magnetic nanoparticles are super-paramagnetic materials with strong magnetic properties, the immobilized cellulase on PAN coated magnetic particles can be easily recovered.

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