Hydrazide-functionalized poly(2-hydroxyethyl methacrylate) microspheres for immobilization of horseradish peroxidase
Hydrazide-functionalized poly(2-hydroxyethyl methacrylate) microspheres for immobilization of horseradish peroxidase
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DOI：
10.1021/bp990006w
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发表时间：
1999-03-01
影响因子：
2.9
通讯作者：
Benes, M
中科院分区：
文献类型：
--
作者：
Horák, D;Karpísek, M;Benes, M
以甲基丙烯酸羟乙酯（HEMA）和二甲基丙烯酸乙二醇酯（EDMA）为原料，通过分散聚合法制备了无孔交联聚（HEMA-co-EDMA）微球。在作为空间稳定剂的乙酸丁酸纤维素和过氧化二苯甲酰引发剂的存在下，在甲苯/2-甲基丙-1-醇中进行聚合。可以通过降低甲苯/2-甲基丙-1-醇比率和通过提高聚合温度来增加粒度。用2，4，6-三氯-1，3，5-三嗪活化微球，使其与己二酰肼结合。在其碳水化合物部分的高碘酸盐氧化后，辣根过氧化物酶被偶联到酰肼官能化的聚（HEMA-co-EDMA）微粒上，高达7.3 μ g酶/g载体，而其活性没有显著损失。固定化过氧化物酶被认为是稳定的，保留超过97%的初始活性时，储存23天后的准备。
Nonporous cross-linked poly(2-hydroxyethyl methacrylate-co-ethylene dimethacrylate) (poly(HEMA-co-EDMA)) microspheres were prepared by dispersion polymerization of HEMA and EDMA. The polymerization was performed in toluene/2-methylpropan-1-ol in the presence of cellulose acetate butyrate as a steric stabilizer and dibenzoyl peroxide initiator. The particle size may be increased by decreasing the toluene/2-methylpropan-1-ol ratio and by increasing polymerization temperature. Adipohydrazide was attached to the microspheres activated with 2,4,6-trichloro-1,3,5-triazine. After periodate oxidation of its carbohydrate moieties, horseradish peroxidase was coupled to the hydrazide-functionalized poly(HEMA-co-EDMA) microparticles up to 7.3 mu g of enzyme/g of carrier without a significant loss of its activity. Immobilized peroxidase was found to be stable, retaining more than 97% of its initial activity when stored for 23 days after the preparation.