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- Title
Enzyme Immobilization for Solid-Phase Catalysis.
- Authors
Fang, Yi; Zhang, Aihua; Li, Shaohua; Sproviero, Michael; Xu, Ming-Qun
- Abstract
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.
- Subjects
ENCAPSULATION (Catalysis); IMMOBILIZED enzymes; DEOXYRIBOZYMES; ENZYMES; PHASE-transfer catalysis
- Publication
Catalysts (2073-4344), 2019, Vol 9, Issue 9, p732
- ISSN
2073-4344
- Publication type
Article
- DOI
10.3390/catal9090732