《蒙脱石共固定化淀粉酶和糖化酶的研究》-毕业论文.doc

PAGE 蒙脱石共固定化淀粉酶和糖化酶的研究 摘要 自20世纪60年代以来,固定化酶技术在食品与发酵工业、有机合成等领域得到了广泛的应用。而近年来酶的共固定化技术,既有固定化酶的高稳定性和连续操作能力,又具有酶之间的协同作用，显示出很强的优越性。 本研究选择目前工业上应用比较广泛的蒙脱石为载体,采用吸附法共固定了α-淀粉酶和糖化酶双酶。实验所用的蒙脱石经提纯钠化后，用KH570对其进行偶联，然后用经以上处理的蒙脱石对双酶同时固定。并通过测定其活性和酶的吸附率分析实验结果，确定了共固定化的最适条件,研究了共固定化酶的性质,为工业应用提供了一定的理论依据。 经本实验所确定的共固定最佳条件为淀粉酶（U）：糖化酶（U）：蒙脱石（g）为15：7.5：0.2，pH为5.5，温度为20℃。在最佳固定条件下共固定化酶的性质为：最适反应温度为55℃左右;最适反应pH为5.5；稳定性在低温下较好，而在高温下不甚理想；共固定化酶在在0℃下保存10天后其相对酶活仍在50%以上，使用四次后其相对酶活仍有50%以上，故储藏性能和操作性能较好。 关键词：共固定化，α-淀粉酶，糖化酶。 Studies on co-immobilization of α-amylase and glucoamylase on montmorillonite Abstract Since the advent of the immobilized enzymes in the 60s of last century, the immobilized enzymes has been widely used in the areas of food and fermentation industry and organic synthesis. The co-immobilized enzymes has both high stability and continuous operation capability and shows a strong advantage. This study has chosen the montmorillonite which was widely used in todays industrial production as the carrier , and fixed α-amylase and glucoamylase dual enzyme system by adsorption . Determined the optimal conditions of co-immobilization and studied the nature of the co-immobilized enzyme, providing a theoretical basis for industrial applications .The montmorillonite used in this experiment has been purified and Na-treated , and then conjugated by one coupling agent which named KH570 in order to improve its adsorption capacity of the enzymes. Then using the montmorillonite to fix two-enzyme. Determined the optimal conditions of co-immobilization and studied the nature of the co-immobilized enzyme, providing a theoretical basis for industrial applications . The present study identified a total of constant optimal conditions for amylase (U): glucoamylase (U): montmorillonite (g) as 15:7.5:0.2, pH 5.5, temperature 20 ℃. In the best condition of the fixed nature of immobilized enzyme: optimal temperature was around 55 ℃; optimal reaction pH 5.5; stability at low temperatures well, but not ideal at hig