1 research outputs found
Enhancing Multistep DNA Processing by Solid-Phase Enzyme Catalysis on Polyethylene Glycol Coated Beads
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