The endoglycosidase activity of Dispersin B is mediated through electrostatic interactions with cationic poly-β-(1→6)-N-acetylglucosamine

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dc.contributor.authorBreslawec, Alexandra P.
dc.contributor.authorWang, Shaochi
dc.contributor.authorMonahan, Kathleen N.
dc.contributor.authorBarry, Lucas L.
dc.contributor.authorPoulin, Myles B.
dc.date.accessioned2023-09-28T19:35:16Z
dc.date.available2023-09-28T19:35:16Z
dc.date.issued2022-09-09
dc.description.abstractBacterial biofilms consist of bacterial cells embedded within a self-produced extracellular polymeric substance (EPS) composed of exopolysaccharides, extra cellular DNA, proteins and lipids. The enzyme Dispersin B (DspB) is a CAZy type 20 β-hexosaminidase enzyme that catalyses the hydrolysis of poly-N-acetylglucosamine (PNAG), a major biofilm polysaccharide produced by a wide variety of biofilm-forming bacteria. Native PNAG is partially de-N-acetylated, and the degree of deacetylation varies between species and dependent on the environment. We have previously shown that DspB is able to perform both endo- and exo-glycosidic bond cleavage of PNAG depending on the de-N-acetylation patterns present in the PNAG substrate. Here, we used a combination of synthetic PNAG substrate analogues, site-directed mutagenesis and in vitro biofilm dispersal assay to investigate the molecular basis for the endo-glycosidic cleavage activity of DspB and the importance of this activity for dispersal of PNAG-dependent Staphylococcus epidermidis biofilms. We found that D242 contributes to the endoglycosidase activity of DspB through electrostatic interactions with cationic substrates in the −2 binding site. A DspBD242N mutant was highly deficient in endoglycosidase activity while maintaining exoglycosidase activity. When used to disperse S. epidermidis biofilms, this DspBD242N mutant resulted in an increase in residual biofilm biomass after treatment when compared to wild-type DspB. These results suggest that the de-N-acetylation of PNAG in S. epidermidis biofilms is not uniformly distributed and that the endoglycosidase activity of DspB is required for efficient biofilm dispersal.
dc.description.urihttps://doi.org/10.1111/febs.16624
dc.identifierhttps://doi.org/10.13016/dspace/rtxt-mpie
dc.identifier.citationBreslawec, A.P., Wang, S., Monahan, K.N., Barry, L.L. and Poulin, M.B. (2023), The endoglycosidase activity of Dispersin B is mediated through electrostatic interactions with cationic poly-β-(1→6)-N-acetylglucosamine. FEBS J, 290: 1049-1059.
dc.identifier.urihttp://hdl.handle.net/1903/30627
dc.language.isoen_US
dc.publisherWiley
dc.relation.isAvailableAtDigital Repository at the University of Marylanden_us
dc.relation.isAvailableAtChemistry & Biochemistryen_us
dc.relation.isAvailableAtCollege of Computer, Mathematical & Natural Sciencesen_us
dc.relation.isAvailableAtUniversity of Maryland (College Park, MD)en_us
dc.subjectbiofilm dispersal
dc.subjectdispersin B
dc.subjectexopolysaccharides
dc.subjectglycoside hydrolase
dc.subjectPNAG
dc.titleThe endoglycosidase activity of Dispersin B is mediated through electrostatic interactions with cationic poly-β-(1→6)-N-acetylglucosamine
dc.typeArticle
local.equitableAccessSubmissionNo

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