Covalent immobilization of enzymes on solid supports provides an alternative approach to homogeneous biocatalysis by adding the benefits of simple enzyme removal, improved stability, and adaptability to automation and high-throughput applications. Nevertheless, immobilized (IM) enzymes generally suffer from reduced activity compared to their soluble counterparts. The nature and hydrophobicity of the supporting material surface can introduce enzyme conformational change, spatial confinement, and limited substrate accessibility, all of which will result in loss of the immobilized enzyme activity. In this work, we demonstrate through kinetic studies that flexible polyethylene glycol (PEG) moieties modifying the surface of magnetic beads improve the activity of covalently immobilized DNA replication enzymes. PEG-modified immobilized enzymes were utilized in library construction for Illumina next-generation sequencing (NGS) increasing the read coverage across AT-rich regions.

中文翻译：

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通过固相酶催化在聚乙二醇包覆的磁珠上增强多步DNA处理

酶在固相支持物上的共价固定化提供了简单的酶去除，提高的稳定性以及对自动化和高通量应用的适应性等优点，为均相生物催化提供了另一种方法。然而，固定化（IM）酶通常比其可溶性对应物活性降低。支撑材料表面的性质和疏水性会引入酶的构象变化，空间限制和有限的底物可及性，所有这些都会导致固定化酶活性的丧失。在这项工作中，我们通过动力学研究证明，修饰磁珠表面的柔性聚乙二醇（PEG）部分可改善共价固定的DNA复制酶的活性。