摘要
脂肪酶（Lipase，EC 3.1.1.3）主要是水解三甘油脂成甘油及脂肪酸。廣泛地應用於脂肪、油品、食品及清潔劑工業。大多數的脂肪酶為適中溫性的酵素，無法水解在常溫下為固體的受質及對化學變性劑較無抗性，因此在工業應用上有所限制。但因其催化作用通常伴隨著高特定選擇和高鏡像選擇性，所以在生物技術上反而擁有巨大發展潛力。因此本實驗利用磁石攪拌器乳化油－水反應液以研究從 Pseudomonas fluorescenes CCRC-17015所的抽取的粗製脂肪酶，在批式反應器內催化甘油三丁酸酯的水解反應速率，並探討基質濃度、溫度、pH值、轉速等因素對水解速率之影響。而為了增加酵素的實用性，本實驗以多孔隙性的甲基丙烯醯胺共聚物為載體，此載體經由乙二胺改質、戊二醛活化後，做為酵素固定化的材質，並利用共價結合的方法，將鍵酵素結以形成固定化酵素。對此，固定化酵素以溫度、pH、轉速等不同的操作條件加以探討，並且計算出Km、Vmax。實驗結果顯示，脂肪酶固定化後擁有良好的熱穩定性與耐鹼性。
並列摘要
The primary purposes of lipase is use to hydrolyze triacylglyceol becomes the glycerol and the fatty acid. This enzyme has been widely used in fat, Food ingredients and Detergents industry. Most of lipases are mesophilic enzyme, which can’t hydrolyze substrate that exists in solid form at room temperature and are unstable when chemical solvents are present. This has severely restricted the application of lipase in industries. But in this catalysis reaction, it came with substrate specificity and enantioselectivity. Therefore, the the lipase is one of the most potential in biotechnological. The purpose of this study is research the hydrolysis rate of tributyrin in batch reactor use crude lipase from Pseudomonas fluorescenes CCRC-17015 and using magnetic stir reactor to emulsify oil-water phase. In order to increase the usability of enzyme, the study use lipase immobilization onto porous polymethyacrylamide by covalent fixation. The carrier modified with ethylenediamine and activated by glutaraldehyde. The primary purposes of this thesis are to study the effect of hydrolysis rate at different substrate concentrations, temperature, pH and stirring speed. Final, to determine the operating conditions of free enzyme and immobilized enzyme then we can get kinetic parameters Km and Vmax values. The experimental results showed lipase after immobilization own good thermal stability and greater stability at higher pH value.
參考文獻
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