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首页> 外文期刊>Biotechnology Progress >protein-Coated Polymer as a Matrix for Enzyme Immobilization: Immobilization of Trypsin on Bovine Serum Albumin-Coated AUyl Glycidyl Ether-Ethylene Glycol Dimethacrylate Copolymer
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protein-Coated Polymer as a Matrix for Enzyme Immobilization: Immobilization of Trypsin on Bovine Serum Albumin-Coated AUyl Glycidyl Ether-Ethylene Glycol Dimethacrylate Copolymer

机译：蛋白涂层的聚合物作为酶固定的基质：胰蛋白酶在牛血​​清白蛋白涂层的AUyl缩水甘油醚-乙二醇二甲基丙烯酸酯共聚物上的固定

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摘要

AUyl glycidyl ether (AGE)-ethylene glycol dimethacrylate (EGDM) copolymer with 25% crosslink density (AGE-25) shows excellent bovine serum albumin (BSA) adsorption (up to 16% (w/w)) at pH 8.0 and the adsorbed BSA is strongly bound. This protein-coated polymer provides a novel matrix with naturally existing functional groups such as thiol, amino, and carboxylic acid that are available for covalent immobilization of functional enzymes. Employing appropriate strategies, trypsin as a model protein was covalently bound to BSA-coated matrix both independently, and in a stepwise manner on the same matrix, with less than 5% loss of enzyme activity during immobilization. Glutaraldehyde crosslinking after immobilization provide stable enzyme preparation with activity of 510 units/g recycled up to six times without loss of enzyme activity. AFM studies reveal that the polymer surface has protein peaks and valleys rather than a uniform monolayer distribution of the protein and the immobilized enzyme preparation can best be described as polymer supported cross-linked enzyme aggregates (CLEAs).
机译：交联密度为25％（AGE-25）的AUyl缩水甘油基醚（AGE）-乙二醇二甲基丙烯酸酯（EGDM）共聚物在pH 8.0时表现出优异的牛血清白蛋白（BSA）吸附（高达16％（w / w）），并且被吸附BSA具有很强的约束力。这种蛋白涂层的聚合物提供了一种新型的基质，该基质具有天然存在的官能团，例如硫醇，氨基和羧酸，可用于共价固定功能酶。通过采用适当的策略，胰蛋白酶作为模型蛋白可以独立地和逐步地共价结合到BSA包被的基质上，并固定在同一基质上，固定过程中酶活损失少于5％。固定后的戊二醛交联可提供稳定的酶制剂，其活性为510单位/ g，最多可循环六次，而不会损失酶的活性。原子力显微镜研究表明，聚合物表面具有蛋白质的峰和谷，而不是蛋白质的均匀单层分布，固定化的酶制剂可以最好地描述为聚合物支持的交联酶聚集体（CLEA）。

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