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含环碳酸酯基固定化酶载体的合成及其性能研究
Syntheses and Properties of the New Novel Supports for Enzymes Containing Cyclic Carbonate
【摘要】 环碳酸酯基具有于温和条件下与氨基反应形成羟基与氨基甲酸酯的特征 .通过反相悬浮聚合 ,以乙烯撑碳酸酯为反应性单体 ,亲水性N ,N′ 亚甲基双丙烯酰胺为交联剂 ,分别选用N 乙烯基吡咯烷酮和丙烯酸 β 羟乙酯两种亲水性共单体为合成酶载体的成分 ,首次以二甲基甲酰胺和线型高分子聚乙二醇 40 0为复合致孔剂合成了两种性能优越的 ,可快速且可保持酶活力的固载胰蛋白酶的大孔型交联高分子酶载体 .其以σ共价键形成的固定化酶和以纯物理吸附形式吸附的酶同时存在于树脂骨架之中 ,根据相似相容原理 ,固载于载体上的酶分子容易吸附溶液中酶分子使其以物理吸附的形式存在 ,后者在经过 6次使用后近乎定量脱落 ,最终保持 85%活力不变者当为σ共价键结合酶 .纯σ共价键结合于高分子骨架的酶不易脱落 ,利用这一特性可使固定化酶的催化过程简化 ,可避免繁杂而冗长的分离脱落酶的过程 .可望将这种高分子酶载体用于临床医学中属于多肽类的分子量为 2 0 0 0左右的中级分子量毒素的无泄露排除 .
【Abstract】 Cyclic carbonate groups can react with amino groups to form hydroxyl groups and carbonate groups under mild conditions.Two kinds of polymers containing cyclic carbonate were obtained by means of anti\|phase suspension polymerization with vinylene carbonate as reactive monomer,N,N′ methylene biacrylamide as crosslinking agent,N\|vinyl\|2\|pyrrolidnone and 2\|hydroxyethyl acrylate as hydrophilic comonomer respectively,and dimethyl formamide and poly(ethylene glycol)( M n=400) as diluent agents.The supports which were synthesized could be used to immobilize trypsin quickly,containing activity.There were two kinds of immobilized trypsins on the supports,one was in the form of σ bond,and the other was in the form of physical adsorption.The latter dropped completely after operating 6 times,but the former didn’t drop easily and contained 85% activity.The immobilization could be simplified by using this method.These supports will be used to remove toxins with molecular weight of 2 000 in clinic medicine.
【Key words】 vinylene carbonate; anti\|phase suspension polymerization; trypsin; immobilized enzyme;
- 【文献出处】 中国生物化学与分子生物学报 ,Chinese Journal of Biochemistry and Molecular Biology , 编辑部邮箱 ,2001年03期
- 【分类号】O631
- 【被引频次】31
- 【下载频次】249