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Functionalized bicomponent polymer membranes as supports for covalent immobilization of enzymes

机译:功能化的双组分聚合物膜作为共价固定酶的载体

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The present work was aimed at developing new reactive polymer materials suitable to be used as supports for enzyme covalent immobilization. Thus, new bicomponent polymer membranes were developed using poly(acrylonitrile-co-vinyl acetate) (PAN-co-PVAc) in mixture with polyvinyl alcohol (PVA). First of all, PAN-co-PVAc/PVA blends were dissolved in DMSO, until a homogenous polymer solution was obtained. To prepare membranes, these solutions were cast on a glass plate followed by the immersion of this plate in a coagulation bath containing a 50%-50% (volume %) water-isopropyl alcohol mixture. In this way membranes containing OH functional groups were obtained. Before tyrosinase immobilization, membranes were functionalized with glutardialdehyde; hence CHO binding sites were inserted and membrane became reactive for the enzyme. In order to prove that both, functionalization and immobilization reactions, were successful, the modifications produced by these reactions were investigated by various techniques i.e. Fourier Transform Infrared Spectrometry, Thermal Gravimetric Analysis, Differential Scanning Calorimetry and Atomic Force Microscopy. The occurrence of important changes in membrane features confirmed the success of both reactions. Furthermore, the activity of bonded enzyme was determined by pyrocatechol method and compared to the activity of the free enzyme. (C) 2015 Elsevier B.V. All rights reserved.
机译:本工作旨在开发适合用作酶共价固定载体的新型反应性聚合物材料。因此,使用聚(丙烯腈-共-乙酸乙烯酯)(PAN-co-PVAc)和聚乙烯醇(PVA)的混合物,开发了新的双组分聚合物膜。首先,将PAN-co-PVAc / PVA共混物溶解在DMSO中,直到获得均质的聚合物溶液。为了制备膜,将这些溶液浇铸在玻璃板上,然后将该板浸入包含50%-50%(体积%)水-异丙醇混合物的凝结浴中。以这种方式,获得了包含OH官能团的膜。在固定酪氨酸酶之前,用戊二醛对膜进行功能化;因此,插入了CHO结合位点,膜对酶产生了反应。为了证明官能化和固定化反应均成功,通过各种技术,即傅立叶变换红外光谱,热重分析,差示扫描量热法和原子力显微镜,研究了这些反应产生的修饰。膜特征的重要变化的发生证实了两个反应的成功。此外,通过邻苯二酚法测定结合酶的活性,并与游离酶的活性进行比较。 (C)2015 Elsevier B.V.保留所有权利。

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