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Polymer-Assisted Biocatalysis: Polyamide 4 Microparticles as Promising Carriers of Enzymatic Function

机译:聚合物辅助生物分析:聚酰胺4微粒作为酶功能的承诺载体

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摘要

This study reports a new strategy for enzyme immobilization based on passive immobilization in neat and magnetically responsive polyamide 4 (PA4) highly porous particles. The microsized particulate supports were synthesized by low-temperature activated anionic ring-opening polymerization. The enzyme of choice was laccase from Trametes versicolor and was immobilized by either adsorption on prefabricated PA4 microparticles (PA4@iL) or by physical in situ entrapment during the PA4 synthesis (PA4@eL). The surface topography of all PA4 particulate supports and laccase conjugates, as well as their chemical and physical structure, were studied by microscopic, spectral, thermal, and synchrotron WAXS/SAXS methods. The laccase content and activity in each conjugate were determined by complementary spectral and enzyme activity measurements. PA4@eL samples displayed >93% enzyme retention after five incubation cycles in an aqueous medium, and the PA4@iL series retained ca. 60% of the laccase. The newly synthesized PA4-laccase complexes were successfully used in dyestuff decolorization aiming at potential applications in effluent remediation. All of them displayed excellent decolorization of positively charged dyestuffs reaching ~100% in 15 min. With negative dyes after 24 h the decolorization reached 55% for PA4@iL and 85% for PA4@eL. A second consecutive decolorization test revealed only a 5–10% decrease in effectiveness indicating the reusability potential of the laccase-PA4 conjugates.
机译:本研究报告了基于纯和磁响应聚酰胺4(PA4)高度多孔颗粒中的被动固定的酶固定化的新策略。通过低温活化的阴离子开环聚合来合成微颗粒载体。选择的酶是来自Trametes Versicolor的漆酶,通过在预制PA4微粒(PA4 @ IL)上的吸附或通过在PA4合成期间的物理挤压来固定(PA4 @ EL)。通过微观,光谱,热和同步蜡蜡/萨克斯方法研究了所有PA4颗粒状支持和漆酶及其化学和物理结构的所有PA4颗粒状蛋白质的表面形貌。通过互补光谱和酶活性测量来确定每个缀合物中的漆酶含量和活性。 PA4 @ EL样品显示>水性介质中五个培养循环后显示> 93%酶保留,PA4 @ IL系列保留了CA. 60%的漆粉。新合成的PA4-漆酶配合物成功用于染色脱色,旨在潜在的污水修复应用。所有这些都显示出优异的带正电荷染料的脱色,15分钟内达到〜100%。 24小时后具有负染料,PA4 @ IL的脱色达到55%,PA4 @ EL的85%达到55%。第二连续脱色测试显示,有效性降低仅为5-10%,表明LACCASE-PA4缀合物的可重用性潜力。

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