Publication

Analysis of the substrate specificity of -L-arabinofuranosidases by DNA sequencer-aided fluorophore-assisted carbohydrate electrophoresis

da Fonseca, M. J. M., Jurak, E., Kataja, K., Master, E. R., Berrin, J-G., Stals, I., Desmet, T., Van Landschoot, A. & Briers, Y., Dec-2018, In : Applied Microbiology and Biotechnology. 102, 23, p. 1009-10102 12 p.

Research output: Contribution to journalArticleAcademicpeer-review

APA

da Fonseca, M. J. M., Jurak, E., Kataja, K., Master, E. R., Berrin, J-G., Stals, I., ... Briers, Y. (2018). Analysis of the substrate specificity of -L-arabinofuranosidases by DNA sequencer-aided fluorophore-assisted carbohydrate electrophoresis. Applied Microbiology and Biotechnology, 102(23), 1009-10102. https://doi.org/10.1007/s00253-018-9389-3

Author

da Fonseca, Maria Joao Mauricio ; Jurak, Edita ; Kataja, Kim ; Master, Emma R. ; Berrin, Jean-Guy ; Stals, Ingeborg ; Desmet, Tom ; Van Landschoot, Anita ; Briers, Yves. / Analysis of the substrate specificity of -L-arabinofuranosidases by DNA sequencer-aided fluorophore-assisted carbohydrate electrophoresis. In: Applied Microbiology and Biotechnology. 2018 ; Vol. 102, No. 23. pp. 1009-10102.

Harvard

da Fonseca, MJM, Jurak, E, Kataja, K, Master, ER, Berrin, J-G, Stals, I, Desmet, T, Van Landschoot, A & Briers, Y 2018, 'Analysis of the substrate specificity of -L-arabinofuranosidases by DNA sequencer-aided fluorophore-assisted carbohydrate electrophoresis' Applied Microbiology and Biotechnology, vol. 102, no. 23, pp. 1009-10102. https://doi.org/10.1007/s00253-018-9389-3

Standard

Analysis of the substrate specificity of -L-arabinofuranosidases by DNA sequencer-aided fluorophore-assisted carbohydrate electrophoresis. / da Fonseca, Maria Joao Mauricio; Jurak, Edita; Kataja, Kim; Master, Emma R.; Berrin, Jean-Guy; Stals, Ingeborg; Desmet, Tom; Van Landschoot, Anita; Briers, Yves.

In: Applied Microbiology and Biotechnology, Vol. 102, No. 23, 12.2018, p. 1009-10102.

Research output: Contribution to journalArticleAcademicpeer-review

Vancouver

da Fonseca MJM, Jurak E, Kataja K, Master ER, Berrin J-G, Stals I et al. Analysis of the substrate specificity of -L-arabinofuranosidases by DNA sequencer-aided fluorophore-assisted carbohydrate electrophoresis. Applied Microbiology and Biotechnology. 2018 Dec;102(23):1009-10102. https://doi.org/10.1007/s00253-018-9389-3


BibTeX

@article{07336e10cbba4e76b2d5cffffbc35a26,
title = "Analysis of the substrate specificity of -L-arabinofuranosidases by DNA sequencer-aided fluorophore-assisted carbohydrate electrophoresis",
abstract = "Carbohydrate-active enzyme discovery is often not accompanied by experimental validation, demonstrating the need for techniques to analyze substrate specificities of carbohydrate-active enzymes in an efficient manner. DNA sequencer-aided fluorophore-assisted carbohydrate electrophoresis (DSA-FACE) is utmost appropriate for the analysis of glycoside hydrolases that have complex substrate specificities. DSA-FACE is demonstrated here to be a highly convenient method for the precise identification of the specificity of different -L-arabinofuranosidases for (arabino)xylo-oligosaccharides ((A)XOS). The method was validated with two -L-arabinofuranosidases (EC 3.2.1.55) with well-known specificity, specifically a GH62 -L-arabinofuranosidase from Aspergillus nidulans (AnAbf62A-m2,3) and a GH43 -L-arabinofuranosidase from Bifidobacterium adolescentis (BaAXH-d3). Subsequently, application of DSA-FACE revealed the AXOS specificity of two -L-arabinofuranosidases with previously unknown AXOS specificities. PaAbf62A, a GH62 -L-arabinofuranosidase from Podospora anserina strain S mat+, was shown to target the O-2 and the O-3 arabinofuranosyl monomers as side chain from mono-substituted -D-xylosyl residues, whereas a GH43 -L-arabinofuranosidase from a metagenomic sample (AGphAbf43) only removes an arabinofuranosyl monomer from the smallest AXOS tested. DSA-FACE excels ionic chromatography in terms of detection limit for (A)XOS (picomolar sensitivity), hands-on and analysis time, and the analysis of the degree of polymerization and binding site of the arabinofuranosyl substituent.",
keywords = "alpha-L-arabinofuranosidases, Substrate specificity, DSA-FACE, HPAEC-PAD, Enzyme analysis, ALPHA-L-ARABINOFURANOSIDASES, INDUCED FLUORESCENCE DETECTION, CAPILLARY-ELECTROPHORESIS, BIFIDOBACTERIUM-ADOLESCENTIS, (1,4)-BETA-D-ARABINOXYLAN ARABINOFURANOHYDROLASE, PODOSPORA-ANSERINA, OLIGOSACCHARIDES, ARABINOXYLAN, GH62, POLYSACCHARIDES",
author = "{da Fonseca}, {Maria Joao Mauricio} and Edita Jurak and Kim Kataja and Master, {Emma R.} and Jean-Guy Berrin and Ingeborg Stals and Tom Desmet and {Van Landschoot}, Anita and Yves Briers",
year = "2018",
month = "12",
doi = "10.1007/s00253-018-9389-3",
language = "English",
volume = "102",
pages = "1009--10102",
journal = "Applied Microbiology and Biotechnology",
issn = "0175-7598",
publisher = "SPRINGER",
number = "23",

}

RIS

TY - JOUR

T1 - Analysis of the substrate specificity of -L-arabinofuranosidases by DNA sequencer-aided fluorophore-assisted carbohydrate electrophoresis

AU - da Fonseca, Maria Joao Mauricio

AU - Jurak, Edita

AU - Kataja, Kim

AU - Master, Emma R.

AU - Berrin, Jean-Guy

AU - Stals, Ingeborg

AU - Desmet, Tom

AU - Van Landschoot, Anita

AU - Briers, Yves

PY - 2018/12

Y1 - 2018/12

N2 - Carbohydrate-active enzyme discovery is often not accompanied by experimental validation, demonstrating the need for techniques to analyze substrate specificities of carbohydrate-active enzymes in an efficient manner. DNA sequencer-aided fluorophore-assisted carbohydrate electrophoresis (DSA-FACE) is utmost appropriate for the analysis of glycoside hydrolases that have complex substrate specificities. DSA-FACE is demonstrated here to be a highly convenient method for the precise identification of the specificity of different -L-arabinofuranosidases for (arabino)xylo-oligosaccharides ((A)XOS). The method was validated with two -L-arabinofuranosidases (EC 3.2.1.55) with well-known specificity, specifically a GH62 -L-arabinofuranosidase from Aspergillus nidulans (AnAbf62A-m2,3) and a GH43 -L-arabinofuranosidase from Bifidobacterium adolescentis (BaAXH-d3). Subsequently, application of DSA-FACE revealed the AXOS specificity of two -L-arabinofuranosidases with previously unknown AXOS specificities. PaAbf62A, a GH62 -L-arabinofuranosidase from Podospora anserina strain S mat+, was shown to target the O-2 and the O-3 arabinofuranosyl monomers as side chain from mono-substituted -D-xylosyl residues, whereas a GH43 -L-arabinofuranosidase from a metagenomic sample (AGphAbf43) only removes an arabinofuranosyl monomer from the smallest AXOS tested. DSA-FACE excels ionic chromatography in terms of detection limit for (A)XOS (picomolar sensitivity), hands-on and analysis time, and the analysis of the degree of polymerization and binding site of the arabinofuranosyl substituent.

AB - Carbohydrate-active enzyme discovery is often not accompanied by experimental validation, demonstrating the need for techniques to analyze substrate specificities of carbohydrate-active enzymes in an efficient manner. DNA sequencer-aided fluorophore-assisted carbohydrate electrophoresis (DSA-FACE) is utmost appropriate for the analysis of glycoside hydrolases that have complex substrate specificities. DSA-FACE is demonstrated here to be a highly convenient method for the precise identification of the specificity of different -L-arabinofuranosidases for (arabino)xylo-oligosaccharides ((A)XOS). The method was validated with two -L-arabinofuranosidases (EC 3.2.1.55) with well-known specificity, specifically a GH62 -L-arabinofuranosidase from Aspergillus nidulans (AnAbf62A-m2,3) and a GH43 -L-arabinofuranosidase from Bifidobacterium adolescentis (BaAXH-d3). Subsequently, application of DSA-FACE revealed the AXOS specificity of two -L-arabinofuranosidases with previously unknown AXOS specificities. PaAbf62A, a GH62 -L-arabinofuranosidase from Podospora anserina strain S mat+, was shown to target the O-2 and the O-3 arabinofuranosyl monomers as side chain from mono-substituted -D-xylosyl residues, whereas a GH43 -L-arabinofuranosidase from a metagenomic sample (AGphAbf43) only removes an arabinofuranosyl monomer from the smallest AXOS tested. DSA-FACE excels ionic chromatography in terms of detection limit for (A)XOS (picomolar sensitivity), hands-on and analysis time, and the analysis of the degree of polymerization and binding site of the arabinofuranosyl substituent.

KW - alpha-L-arabinofuranosidases

KW - Substrate specificity

KW - DSA-FACE

KW - HPAEC-PAD

KW - Enzyme analysis

KW - ALPHA-L-ARABINOFURANOSIDASES

KW - INDUCED FLUORESCENCE DETECTION

KW - CAPILLARY-ELECTROPHORESIS

KW - BIFIDOBACTERIUM-ADOLESCENTIS

KW - (1,4)-BETA-D-ARABINOXYLAN ARABINOFURANOHYDROLASE

KW - PODOSPORA-ANSERINA

KW - OLIGOSACCHARIDES

KW - ARABINOXYLAN

KW - GH62

KW - POLYSACCHARIDES

U2 - 10.1007/s00253-018-9389-3

DO - 10.1007/s00253-018-9389-3

M3 - Article

VL - 102

SP - 1009

EP - 10102

JO - Applied Microbiology and Biotechnology

JF - Applied Microbiology and Biotechnology

SN - 0175-7598

IS - 23

ER -

ID: 72808586