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

Tutkimustuotos: Lehtiartikkeli

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

julkaisussa: Applied Microbiology and Biotechnology, Vuosikerta 102, Nro 23, 12.2018, s. 10091–10102.

Tutkimustuotos: Lehtiartikkeli

Harvard

Maurício da Fonseca, MJ, Jurak, E, Kataja, K, Master, ER, Berrin, JG, Stals, I, Desmet, T, Van Landschoot, A & Briers, Y 2018, 'Analysis of the substrate specificity of α-L-arabinofuranosidases by DNA sequencer-aided fluorophore-assisted carbohydrate electrophoresis', Applied Microbiology and Biotechnology, Vuosikerta. 102, Nro 23, Sivut 10091–10102. https://doi.org/10.1007/s00253-018-9389-3

APA

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Author

Maurício da Fonseca, Maria João ; 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 α-L-arabinofuranosidases by DNA sequencer-aided fluorophore-assisted carbohydrate electrophoresis. Julkaisussa: Applied Microbiology and Biotechnology. 2018 ; Vuosikerta 102, Nro 23. Sivut 10091–10102.

Bibtex - Lataa

@article{599f28fe7b354535b06df684c619f440,
title = "Analysis of the substrate specificity of α-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 α-L-arabinofuranosidases for (arabino)xylo-oligosaccharides ((A)XOS). The method was validated with two α-L-arabinofuranosidases (EC 3.2.1.55) with well-known specificity, specifically a GH62 α-L-arabinofuranosidase from Aspergillus nidulans (AnAbf62A-m2,3) and a GH43 α-L-arabinofuranosidase from Bifidobacterium adolescentis (BaAXH-d3). Subsequently, application of DSA-FACE revealed the AXOS specificity of two α-L-arabinofuranosidases with previously unknown AXOS specificities. PaAbf62A, a GH62 α-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 β-D-xylosyl residues, whereas a GH43 α-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 = "DSA-FACE, Enzyme analysis, HPAEC-PAD, Substrate specificity, α-L-arabinofuranosidases",
author = "{Maur{\'i}cio da Fonseca}, {Maria Jo{\~a}o} and Edita Jurak and Kim Kataja and Master, {Emma R.} and Berrin, {Jean Guy} and Ingeborg Stals and Tom Desmet and {Van Landschoot}, Anita and Yves Briers",
note = "The research has been financially supported by the research fund of the University College Ghent and Ghent University (B/13845/ 01 ‘ HS Annotatie enzymen",
year = "2018",
month = "12",
doi = "10.1007/s00253-018-9389-3",
language = "English",
volume = "102",
pages = "10091–10102",
journal = "Applied Microbiology & Biotechnology",
issn = "0175-7598",
number = "23",

}

RIS - Lataa

TY - JOUR

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

AU - Maurício da Fonseca, Maria João

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

N1 - The research has been financially supported by the research fund of the University College Ghent and Ghent University (B/13845/ 01 ‘ HS Annotatie enzymen

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 α-L-arabinofuranosidases for (arabino)xylo-oligosaccharides ((A)XOS). The method was validated with two α-L-arabinofuranosidases (EC 3.2.1.55) with well-known specificity, specifically a GH62 α-L-arabinofuranosidase from Aspergillus nidulans (AnAbf62A-m2,3) and a GH43 α-L-arabinofuranosidase from Bifidobacterium adolescentis (BaAXH-d3). Subsequently, application of DSA-FACE revealed the AXOS specificity of two α-L-arabinofuranosidases with previously unknown AXOS specificities. PaAbf62A, a GH62 α-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 β-D-xylosyl residues, whereas a GH43 α-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 α-L-arabinofuranosidases for (arabino)xylo-oligosaccharides ((A)XOS). The method was validated with two α-L-arabinofuranosidases (EC 3.2.1.55) with well-known specificity, specifically a GH62 α-L-arabinofuranosidase from Aspergillus nidulans (AnAbf62A-m2,3) and a GH43 α-L-arabinofuranosidase from Bifidobacterium adolescentis (BaAXH-d3). Subsequently, application of DSA-FACE revealed the AXOS specificity of two α-L-arabinofuranosidases with previously unknown AXOS specificities. PaAbf62A, a GH62 α-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 β-D-xylosyl residues, whereas a GH43 α-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 - DSA-FACE

KW - Enzyme analysis

KW - HPAEC-PAD

KW - Substrate specificity

KW - α-L-arabinofuranosidases

UR - http://www.scopus.com/inward/record.url?scp=85054188828&partnerID=8YFLogxK

U2 - 10.1007/s00253-018-9389-3

DO - 10.1007/s00253-018-9389-3

M3 - Article

VL - 102

SP - 10091

EP - 10102

JO - Applied Microbiology & Biotechnology

JF - Applied Microbiology & Biotechnology

SN - 0175-7598

IS - 23

ER -

ID: 28704629