Vis enkel innførsel

dc.contributor.authorDe Santi, Concetta
dc.contributor.authorWillassen, Nils Peder
dc.contributor.authorWilliamson, Adele Kim
dc.date.accessioned2017-03-15T14:33:52Z
dc.date.available2017-03-15T14:33:52Z
dc.date.issued2016
dc.description.abstractBackground The glucuronoyl esterase enzymes of wood-degrading fungi (Carbohydrate Esterase family 15; CE15) form part of the hemicellulolytic and cellulolytic enzyme systems that break down plant biomass, and have possible applications in biotechnology. Homologous enzymes are predicted in the genomes of several bacteria, however these have been much less studied than their fungal counterparts. Here we describe the recombinant production and biochemical characterization of a bacterial CE15 enzyme denoted MZ0003, which was identified by in silico screening of a prokaryotic metagenome library derived from marine Arctic sediment. MZ0003 has high similarity to several uncharacterized gene products of polysaccharide-degrading bacterial species, and phylogenetic analysis indicates a deep evolutionary split between these CE15s and fungal homologs. Results MZ0003 appears to differ from previously-studied CE15s in some aspects. Some glucuronoyl esterase activity could be measured by qualitative thin-layer chromatography which confirms its assignment as a CE15, however MZ0003 can also hydrolyze a range of other esters, including p-nitrophenyl acetate, which is not acted upon by some fungal homologs. The structure of MZ0003 also appears to differ as it is predicted to have several large loop regions that are absent in previously studied CE15s, and a combination of homology-based modelling and site-directed mutagenesis indicate its catalytic residues deviate from the conserved Ser-His-Glu triad of many fungal CE15s. Taken together, these results indicate that potentially unexplored diversity exists among bacterial CE15s, and this may be accessed by investigation of the microbial metagenome. The combination of low activity on typical glucuronoyl esterase substrates, and the lack of glucuronic acid esters in the marine environment suggest that the physiological substrate of MZ0003 and its homologs is likely to be different from that of related fungal enzymes.en_US
dc.descriptionPublished version. Source at <a href=http://doi.org/10.1371/journal.pone.0159345>http://doi.org/10.1371/journal.pone.0159345</a>. License <a href=https://creativecommons.org/licenses/by/4.0/>CC BY 4.0</a>.en_US
dc.identifier.citationDe Santi C, Willassen NP, Williamson AK. Biochemical characterization of a family 15 carbohydrate esterase from a bacterial marine Arctic metagenome. PLoS ONE. 2016;11:e0159345(7)en_US
dc.identifier.issn1932-6203
dc.identifier.otherFRIDAID 1412926
dc.identifier.other10.1371/journal.pone.0159345
dc.identifier.urihttp://hdl.handle.net/10037/10715
dc.language.isoengen_US
dc.publisherPublic Library of Scienceen_US
dc.relation.journalPLoS ONE
dc.rights.accessRightsopenAccessen_US
dc.subjectEsterasesen_US
dc.subjectSequence alignmenten_US
dc.subjectEstersen_US
dc.subjectMarine bacteriaen_US
dc.subjectFungal structureen_US
dc.subjectPhylogenetic analysisen_US
dc.subjectMultiple alignment calculationen_US
dc.subjectEnzymesen_US
dc.subjectVDP::Mathematics and natural science: 400::Basic biosciences: 470en_US
dc.titleBiochemical characterization of a family 15 carbohydrate esterase from a bacterial marine Arctic metagenomeen_US
dc.typeJournal articleen_US
dc.typeTidsskriftartikkelen_US
dc.typePeer revieweden_US


Tilhørende fil(er)

Denne innførselen finnes i følgende samling(er)

Vis enkel innførsel