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Lookup NU author(s): Dr Claire Dumon, Professor William WillatsORCiD
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License (CC BY-NC 4.0).
Carbohydrate-active enzymes have multiple biological roles and industrial applications. Advances in genome and transcriptome sequencing together with associated bioinformatics tools have identified vast numbers of putative carbohydrate-degrading and -modifying enzymes including glycoside hydrolases and lytic polysaccharide monooxygenases. However, there is a paucity of methods for rapidly screening the activities of these enzymes. By combining the multiplexing capacity of carbohydrate microarrays with the specificity of molecular probes, we have developed a sensitive, high throughput, and versatile semiquantitative enzyme screening technique that requires low amounts of enzyme and substrate. The method can be used to assess the activities of single enzymes, enzyme mixtures, and crude culture broths against single substrates, substrate mixtures, and biomass samples. Moreover, we show that the technique can be used to analyze both endo-acting and exo-acting glycoside hydrolases, polysaccharide lyases, carbohydrate esterases, and lytic polysaccharide monooxygenases. We demonstrate the potential of the technique by identifying the substrate specificities of purified uncharacterized enzymes and by screening enzyme activities from fungal culture broths.
Author(s): Vidal-Melgosa S, Pedersen HL, Schückel J, Arnal G, Dumon C, Amby DB, Monrad RN, Westereng B, Willats WG
Publication type: Article
Publication status: Published
Journal: The Journal of Biological Chemistry
Year: 2015
Volume: 290
Issue: 14
Pages: 9020-9036
Print publication date: 03/04/2015
Online publication date: 03/04/2015
Acceptance date: 05/02/2015
Date deposited: 17/02/2017
ISSN (electronic): 1083-351X
Publisher: American Society for Biochemistry and Molecular Biology, Inc.
URL: http://dx.doi.org/10.1074/jbc.M114.630673
DOI: 10.1074/jbc.M114.630673
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