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尼龙亲和膜的制备条件优化及其表征

作者：聂华丽¹，²，陈天翔²，朱利民 ²

单位： 1. 东华大学生态纺织教育部重点实验室,2. 东华大学

关键词：亲和膜色谱；壳聚糖；木瓜蛋白酶；半胱氨酸蛋白酶抑制剂

DOI号：

分类号： TQ028.8

出版年,卷(期):页码： 2011,31(2):74-77

摘要：

尼龙膜经稀盐酸水解、壳聚糖改性后，以木瓜蛋白酶为亲和膜的配基，通过戊二醛的活化处理后采用共价结合的方法将配基键合在尼龙膜上，从而得到有特异吸附性能的尼龙亲和膜.本实验考察了制备尼龙亲和膜的交联剂戊二醛的质量分数、pH值、温度、反应时间和酶用量对亲和膜上木瓜蛋白酶活力的影响，确定了最适合的制备条件：pH 9.0，质量分数为0.5%的戊二醛，反应温度为45℃，反应时间为6 h，酶用量为10 mg/mL。该优化条件下制备的尼龙亲和膜具备优良的色谱性能，可用于分离纯化半胱氨酸蛋白酶抑制剂.

Nylon membranes are first hydrolyed with dilute HCl and then treated with chitosan before being used as the affinity carrier. Papain as a ligand has been immobilized on the actived membranes with glutaraldehyde as crosslinking agent. The factors involving with the activity of immobilized papain, such as concentration of glutaraldehyde, pH, temperature, reaction time, and the amount of added papain have been studied. The results show that the optimum conditions for papain immobilization are as follows: pH 9.0, 0.5% glutaraldehyde solution, the concentration of added papain is 10 mg/mL, the reaction time is 6 h at 45℃. The nylon membrane prepared is then used to purify cystatin from potato juice, and showed that they are high efficiency affinity membranes base for cystatin separation.

基金项目：

国家自然科学基金（50773009）；教育部高校博士点新教师基金（20090075120001）；中央高校基本科研业务费专项基金

作者简介：

聂华丽（1978-）, 女，上海市人，博士，讲师，从事蛋白质分离与纯化研究，电话：021-67792659，E-Mail：niehuali2004@126.com

参考文献：

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