胰蛋白酶的固定化及其水解酪蛋白的研究

生物工程论文编号:SW131  论文字数:10553,页数:23

目 录
中文摘要
英文摘要…………………………………………………………………….…..…Ⅱ
目录…………………………………………………………………………..…….Ⅲ
绪论………………………………………………………………………..……1
 1.1 胰蛋白酶的研究概述……………………………………………….……1
 1.1.3 胰蛋白酶的制备方法………………………………………………2
 1.2 固定化…………………………………………………………………….4
 1.3 胰蛋白酶的固定化研究现状………………………………………….…6
 1.4  本文的研究思路和目标……………………………………………...…8
2. 实验部分……………………………………………………………………...…9
 2.1 用树脂作介质固定化…...……………………………………………..…9
 2.2 以海藻酸钙为载体固定化胰蛋白酶…………………………………..14
3. 结果与讨论………………………………………….……………………..….18
 3.1实验结果………………………………………...……………...……….18
 3.2实验讨论……………………………………………………...…………18
4. 总结与展望……………………….………………………………………...…19
致谢………………………………….…………………………………………….20
参考文献……………………………………….………………………………….21

摘要：胰蛋白酶是一种动物来源的蛋白水解酶，分子量为23.8 kD，在医药、食品、工业等领域有广泛的应用。本文研究了胰蛋白酶的固定化方法以及利用固定化胰蛋白水解酪蛋白的过程和相关工艺。论文在建立测定胰蛋白酶活力方法的基础上，采用吸附法筛选出适合固定化胰蛋白酶的大孔弱阴离子交换树脂Duolite A7，优化的树脂添加量为0.5g/10mg 蛋白，固定化催化5批次反应后仍有30％以上的酶活力； pH、温度、包埋量等条件对海藻酸钙固定化胰蛋白酶活力的影响进行了考察，海藻酸钠的浓度为15 g/L，CaCl2的浓度为20 g/L，胰蛋白酶的包埋量为60mg/10mL海藻酸钠，反应最适的温度和pH分别为60℃和7.4。
关键词：胰蛋白酶；固定化；海藻酸钙；Duolite A7树脂
 

Abstract: The trypsin that the molecular weight is 23.8 kD is the hydrolytic enzyme which comes from animal. It has the extensive application in medicine, food, industry and so on. This article has studied the trypsin immobilization method as well as the use of trypsin immobilization for casein hydrolysis. Based on determination of trypsin enzyme activity, a macropore weak anion exchange resin Duolite A7 was screened for trypsin immobilization, the optimized resin addition amount is 0.5g/10mg protein, and enzyme activity still reached more than 30% after five-time recycling of the immobilized enzyme. Conditions on pH, temperature, entrapment amount on effect of trypsin activity were investigated by Ca-alginate gel beads, the sodium alginate concentration is 15 g/L, CaCl2 20 g/L, the trypsin entrapment amount is 60mg/10mL sodium alginate; the optimal temperature and pH are 60℃ and 7.4 respectively.
Keywords: Trypsin; Immobilizition; Ca-alginate; Duolite A7 resin