A new versatile microarray-based method for high-throughput screening of carbohydrate-active enzymes

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningfagfællebedømt

Standard

A new versatile microarray-based method for high-throughput screening of carbohydrate-active enzymes. / Vidal Melgosa, Silvia; Pedersen, Henriette Lodberg; Schückel, Julia; Arnal, Grégory; Dumon, Claire; Amby, Daniel Buchvaldt; Monrad, Rune Nygaard; Westereng, Bjørge; Willats, William George Tycho.

I: Journal of Biological Chemistry, Bind 290, 2015, s. 9020-9036.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningfagfællebedømt

Harvard

Vidal Melgosa, S, Pedersen, HL, Schückel, J, Arnal, G, Dumon, C, Amby, DB, Monrad, RN, Westereng, B & Willats, WGT 2015, 'A new versatile microarray-based method for high-throughput screening of carbohydrate-active enzymes', Journal of Biological Chemistry, bind 290, s. 9020-9036. https://doi.org/10.1074/jbc.M114.630673

APA

Vidal Melgosa, S., Pedersen, H. L., Schückel, J., Arnal, G., Dumon, C., Amby, D. B., ... Willats, W. G. T. (2015). A new versatile microarray-based method for high-throughput screening of carbohydrate-active enzymes. Journal of Biological Chemistry, 290, 9020-9036. https://doi.org/10.1074/jbc.M114.630673

Vancouver

Vidal Melgosa S, Pedersen HL, Schückel J, Arnal G, Dumon C, Amby DB o.a. A new versatile microarray-based method for high-throughput screening of carbohydrate-active enzymes. Journal of Biological Chemistry. 2015;290:9020-9036. https://doi.org/10.1074/jbc.M114.630673

Author

Vidal Melgosa, Silvia ; Pedersen, Henriette Lodberg ; Schückel, Julia ; Arnal, Grégory ; Dumon, Claire ; Amby, Daniel Buchvaldt ; Monrad, Rune Nygaard ; Westereng, Bjørge ; Willats, William George Tycho. / A new versatile microarray-based method for high-throughput screening of carbohydrate-active enzymes. I: Journal of Biological Chemistry. 2015 ; Bind 290. s. 9020-9036.

Bibtex

@article{9e125311041748338188b708e630ae21,
title = "A new versatile microarray-based method for high-throughput screening of carbohydrate-active enzymes",
abstract = "Carbohydrate-active enzymes have multiple biological roles and industrial applications. Advances in genome and transcriptome sequencing, together with associated bioinformatic 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 semi-quantitative enzyme-screening technique which requires low amounts of enzyme and substrate. The method can be used to assess the activities of single enzymes, enzyme cocktails and crude culture broths against single substrates, substrate mixtures and biomass samples. Moreover, we show that the technique can be used to analyse 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 un-characterised enzymes and by screening enzyme activities from fungal culture broths.",
author = "{Vidal Melgosa}, Silvia and Pedersen, {Henriette Lodberg} and Julia Sch{\"u}ckel and Gr{\'e}gory Arnal and Claire Dumon and Amby, {Daniel Buchvaldt} and Monrad, {Rune Nygaard} and Bj{\o}rge Westereng and Willats, {William George Tycho}",
note = "Copyright {\circledC} 2015, The American Society for Biochemistry and Molecular Biology.",
year = "2015",
doi = "10.1074/jbc.M114.630673",
language = "English",
volume = "290",
pages = "9020--9036",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology, Inc.",

}

RIS

TY - JOUR

T1 - A new versatile microarray-based method for high-throughput screening of carbohydrate-active enzymes

AU - Vidal Melgosa, Silvia

AU - Pedersen, Henriette Lodberg

AU - Schückel, Julia

AU - Arnal, Grégory

AU - Dumon, Claire

AU - Amby, Daniel Buchvaldt

AU - Monrad, Rune Nygaard

AU - Westereng, Bjørge

AU - Willats, William George Tycho

N1 - Copyright © 2015, The American Society for Biochemistry and Molecular Biology.

PY - 2015

Y1 - 2015

N2 - Carbohydrate-active enzymes have multiple biological roles and industrial applications. Advances in genome and transcriptome sequencing, together with associated bioinformatic 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 semi-quantitative enzyme-screening technique which requires low amounts of enzyme and substrate. The method can be used to assess the activities of single enzymes, enzyme cocktails and crude culture broths against single substrates, substrate mixtures and biomass samples. Moreover, we show that the technique can be used to analyse 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 un-characterised enzymes and by screening enzyme activities from fungal culture broths.

AB - Carbohydrate-active enzymes have multiple biological roles and industrial applications. Advances in genome and transcriptome sequencing, together with associated bioinformatic 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 semi-quantitative enzyme-screening technique which requires low amounts of enzyme and substrate. The method can be used to assess the activities of single enzymes, enzyme cocktails and crude culture broths against single substrates, substrate mixtures and biomass samples. Moreover, we show that the technique can be used to analyse 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 un-characterised enzymes and by screening enzyme activities from fungal culture broths.

U2 - 10.1074/jbc.M114.630673

DO - 10.1074/jbc.M114.630673

M3 - Journal article

VL - 290

SP - 9020

EP - 9036

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

ER -

ID: 131740828