Abstract: With N-isopropyl acrylamide (NIPAM) as temperature sensitive monomer，using the poly (methylacrylic acid - beta hydroxy ethyl ester) (PHEMA) or ethylenediamine aminated poly (methylacrylic acid - beta hydroxy ethyl ester) (PAEMA) as linear polymer，the thermosensitive porous semi-IPN hydrogels were prepared and then modified with maleic anhydride to get two new enzyme immobilization carriersⅠandⅡ，It was found that the pore sizes of the gels were more than 10 microns. With carrierⅠandⅡ，based on the thiol-based click reaction，the immobilization of lipase/ glucoamylase was studied. The results showed that the recovery of immobilized enzymes activity was up to 6.03%，5 times as glutaraldehyde immobilization with high stability.

Key words: temperature sensitive, semi-IPN hydrogels, thiol-based click reaction, immobilized enzyme

摘要： 以N-异丙基丙烯酰胺（NIPAM）为温敏单体，引入线性聚甲基丙烯酸-β-羟乙酯（PHEMA）或乙二胺胺化的聚甲基丙烯酸-β-羟乙酯（PAEMA），制备多孔半互穿水凝胶，并引入马来酰亚胺基团修饰，制备了两种新型水凝胶酶固定化载体Ⅰ和Ⅱ，凝胶的平均孔洞大小均约在10 ?m以上。以Ⅰ和Ⅱ为载体，马来酰亚胺与巯基的点击反应为基础，进行脂肪酶与糖化酶的固定化研究。结果表明，通过点击反应固定化得到的固定化脂肪酶最高活力回收率达6.03%，是相同条件下戊二醛固定化的5倍左右，且稳定性较高。

关键词: 温度敏感, 半互穿水凝胶, 点击反应, 固定化酶