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dc.contributor.authorLeth, Maria Louise
dc.contributor.authorEjby, Morten
dc.contributor.authorMadland, Eva
dc.contributor.authorKitaoku, Yoshihito
dc.contributor.authorSlotboom, Dirk J.
dc.contributor.authorGuskov, Albert
dc.contributor.authorAachmann, Finn Lillelund
dc.contributor.authorHachem, Maher Abou
dc.date.accessioned2020-04-07T12:27:45Z
dc.date.available2020-04-07T12:27:45Z
dc.date.created2019-12-16T11:10:10Z
dc.date.issued2019
dc.identifier.issn1742-464X
dc.identifier.urihttps://hdl.handle.net/11250/2650663
dc.description.abstractEfficient capture of glycans, the prime metabolic resources in the human gut, confers a key competitive advantage for gut microbiota members equipped with extracellular glycoside hydrolases (GHs) to target these substrates. The association of glycans to the bacterial cell surface is typically mediated by carbohydrate binding modules (CBMs). Here, we report the structure of RiCBM86 appended to a GH family 10 xylanase from Roseburia intestinalis. This CBM represents a new family of xylan binding CBMs present in xylanases from abundant and prevalent healthy human gut Clostridiales. RiCBM86 adopts a canonical β‐sandwich fold, but shows structural divergence from known CBMs. The structure of RiCBM86 has been determined with a bound xylohexaose, which revealed an open and shallow binding site. RiCBM86 recognizes only a single xylosyl ring with direct hydrogen bonds. This mode of recognition is unprecedented amongst previously reported xylan binding type‐B CBMs that display more extensive hydrogen‐bonding patterns to their ligands or employ Ca2+ to mediate ligand‐binding. The architecture of RiCBM86 is consistent with an atypically low binding affinity (KD about 0.5 mm for xylohexaose) compared to most xylan binding CBMs. Analyses using NMR spectroscopy corroborated the observations from the complex structure and the preference of RiCBM86 to arabinoxylan over glucuronoxylan, consistent with the largely negatively charged surface flanking the binding site. Mutational analysis and affinity electrophoresis established the importance of key binding residues, which are conserved in the family. This study provides novel insight into the structural features that shape low‐affinity CBMs that mediate extended bacterial glycan capture in the human gut niche.en_US
dc.language.isoengen_US
dc.publisherWileyen_US
dc.titleMolecular insight into a new low-affinity xylan binding module from the xylanolytic gut symbiont Roseburia intestinalisen_US
dc.typePeer revieweden_US
dc.typeJournal articleen_US
dc.description.versionacceptedVersionen_US
dc.source.journalThe FEBS Journalen_US
dc.identifier.doi10.1111/febs.15117
dc.identifier.cristin1761075
dc.description.localcode"Locked until 6.11.2020 due to copyright restrictions. This is the peer reviewed version of an article, which has been published in final form at [https://doi.org/10.1111/febs.15117]. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving. "en_US
cristin.unitcode194,66,15,0
cristin.unitnameInstitutt for bioteknologi og matvitenskap
cristin.ispublishedtrue
cristin.fulltextpreprint
cristin.qualitycode2


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