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

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dc.contributor Aalto-yliopisto fi
dc.contributor Aalto University en
dc.contributor.author Maurício da Fonseca, Maria João
dc.contributor.author Jurak, Edita
dc.contributor.author Kataja, Kim
dc.contributor.author Master, Emma R.
dc.contributor.author Berrin, Jean Guy
dc.contributor.author Stals, Ingeborg
dc.contributor.author Desmet, Tom
dc.contributor.author Van Landschoot, Anita
dc.contributor.author Briers, Yves
dc.date.accessioned 2019-01-14T09:21:18Z
dc.date.available 2019-01-14T09:21:18Z
dc.date.issued 2018-12
dc.identifier.citation Maurício da Fonseca , M J , Jurak , E , Kataja , K , Master , E R , Berrin , J G , 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 , vol. 102 , no. 23 , pp. 10091–10102 . DOI: 10.1007/s00253-018-9389-3 en
dc.identifier.issn 0175-7598
dc.identifier.issn 1432-0614
dc.identifier.other PURE UUID: 599f28fe-7b35-4535-b06d-f684c619f440
dc.identifier.other PURE ITEMURL: https://research.aalto.fi/en/publications/analysis-of-the-substrate-specificity-of-larabinofuranosidases-by-dna-sequenceraided-fluorophoreassisted-carbohydrate-electrophoresis(599f28fe-7b35-4535-b06d-f684c619f440).html
dc.identifier.other PURE LINK: http://www.scopus.com/inward/record.url?scp=85054188828&partnerID=8YFLogxK
dc.identifier.uri https://aaltodoc.aalto.fi/handle/123456789/35957
dc.description The research has been financially supported by the research fund of the University College Ghent and Ghent University (B/13845/ 01 ‘ HS Annotatie enzymen
dc.description.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. en
dc.format.extent 10091–10102
dc.language.iso en en
dc.relation.ispartofseries Applied Microbiology and Biotechnology en
dc.relation.ispartofseries Volume 102, issue 23 en
dc.rights embargoedAccess en
dc.subject.other Biotechnology en
dc.subject.other Applied Microbiology and Biotechnology en
dc.subject.other 1182 Biochemistry, cell and molecular biology en
dc.title Analysis of the substrate specificity of α-L-arabinofuranosidases by DNA sequencer-aided fluorophore-assisted carbohydrate electrophoresis en
dc.type A1 Alkuperäisartikkeli tieteellisessä aikakauslehdessä fi
dc.description.version Peer reviewed en
dc.contributor.department Ghent University
dc.contributor.department Department of Bioproducts and Biosystems
dc.contributor.department Institut National de la Recherche Agronomique
dc.subject.keyword DSA-FACE
dc.subject.keyword Enzyme analysis
dc.subject.keyword HPAEC-PAD
dc.subject.keyword Substrate specificity
dc.subject.keyword α-L-arabinofuranosidases
dc.subject.keyword Biotechnology
dc.subject.keyword Applied Microbiology and Biotechnology
dc.subject.keyword 1182 Biochemistry, cell and molecular biology
dc.identifier.urn URN:NBN:fi:aalto-201901141140
dc.identifier.doi 10.1007/s00253-018-9389-3
dc.date.embargo info:eu-repo/date/embargoEnd/2019-12-31


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