半乳糖氧化酶在珠状和磁性珠状纤维素以及聚（甲基丙烯酸羟乙酯 - 二甲基丙烯酸乙二酯）和磁性聚（甲基丙烯酸羟乙酯 - 二甲基丙烯酸乙二酯）微球上的定向固定化。

Oriented immobilization of galactose oxidase to bead and magnetic bead cellulose and poly(HEMA-co-EDMA) and magnetic poly(HEMA-co-EDMA) microspheres.

作者信息

Bílková Zuzana, Slováková Marcela, Lycka Antonín, Horák Daniel, Lenfeld Jirí, Turková Jaroslava, Churácek Jaroslav

机构信息

Department of Biological and Biochemical Sciences, University of Pardubice, Czech Republic.

出版信息

J Chromatogr B Analyt Technol Biomed Life Sci. 2002 Apr 25;770(1-2):25-34. doi: 10.1016/s0378-4347(01)00439-x.

DOI:10.1016/s0378-4347(01)00439-x

PMID:12013233

Abstract

In order to obtain an active and stable oxidation reactor for daily use in biochemical laboratory we decided to immobilize galactose oxidase orientedly through a carbohydrate chain to the magnetic carriers. We used hydrazide derivatives of non-magnetic and magnetic bead cellulose and of magnetic and non-magnetic poly(HEMA-co-EDMA) microspheres. Activation of the enzyme molecules was done by sodium periodate in the presence of supplements (fucose, CuSO4, catalase). Orientedly immobilized galactose oxidase presents high storage stability and lower susceptibility to inappropriate microenvironmental conditions. Reactor reactivated by three pulses of D-galactose retained practically 100% of its native activity after 6 months. The positive properties of both magnetic carriers were entirely confirmed.

摘要

为了获得一个可在生化实验室日常使用的活性稳定的氧化反应器，我们决定通过碳水化合物链将半乳糖氧化酶定向固定到磁性载体上。我们使用了非磁性和磁性珠状纤维素以及磁性和非磁性聚（甲基丙烯酸羟乙酯 - 二甲基丙烯酸乙二醇酯）微球的酰肼衍生物。在补充剂（岩藻糖、硫酸铜、过氧化氢酶）存在的情况下，用高碘酸钠对酶分子进行活化。定向固定的半乳糖氧化酶具有高储存稳定性，并且对不适当的微环境条件敏感性较低。用三个D - 半乳糖脉冲重新激活的反应器在6个月后实际上保留了其天然活性的100％。两种磁性载体的积极特性都得到了充分证实。