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Enzyme Immobilization for Solid-Phase Catalysis
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DOI：10.3390/catal9090732.png)
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Abstract
En 中文
The covalent immobilization of an enzyme to a solid support can broaden its applicability in various workflows. Immobilized enzymes facilitate catalyst re-use, adaptability to automation or high-throughput applications and removal of the enzyme without heat inactivation or reaction purification. In this report, we demonstrate a step-by-step procedure to carry out the bio-orthogonal immobilization of DNA modifying enzymes employing the self-labelling activity of the SNAP-tag to covalently conjugate the enzyme of interest to the solid support. We also demonstrate how modifying the surface functionality of the support can improve the activity of the immobilized enzyme. Finally, the utility of immobilized DNA-modifying enzymes is depicted through sequential processing of genomic DNA libraries for Illumina next-generation sequencing (NGS), resulting in improved read coverage across AT-rich sequences.
Keywords:
enzyme immobilization
covalent conjugation
DNA modifying enzyme
T4 DNA ligase
magnetic beads
SNAP-tag
polyethylene glycol
hydrophilic polymer
next-generation sequencing
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