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Open AccessArticle

Burkholderia cenocepacia H111 Produces a Water-Insoluble Exopolysaccharide in Biofilm: Structural Determination and Molecular Modelling

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Department of Life Sciences, University of Trieste, via L. Giorgieri 1, Bdg C11, 34127 Trieste, Italy
2
Department of Food Science, Cornell University, Ithaca, NY 14853, USA
3
Department of Chemistry, University of Cape Town, Rondebosch 7701, South Africa
4
Costerton Biofilm Center, Department of Immunology and Microbiology, University of Copenhagen, DK-2200 Copenhagen, Denmark
*
Author to whom correspondence should be addressed.
Int. J. Mol. Sci. 2020, 21(5), 1702; https://doi.org/10.3390/ijms21051702
Received: 11 February 2020 / Revised: 27 February 2020 / Accepted: 28 February 2020 / Published: 2 March 2020
Biofilms are a multicellular way of life, where bacterial cells are close together and embedded in a hydrated macromolecular matrix which offers a number of advantages to the cells. Extracellular polysaccharides play an important role in matrix setup and maintenance. A water-insoluble polysaccharide was isolated and purified from the biofilm produced by Burkholderia cenocepacia strain H111, a cystic fibrosis pathogen. Its composition and glycosidic linkages were determined using Gas–Liquid Chromatography–Mass Spectrometry (GLC–MS) on appropriate carbohydrate derivatives while its complete structure was unraveled by 1D and 2D NMR spectroscopy in deuterated sodium hydroxide (NaOD) aqueous solutions. All the collected data demonstrated the following repeating unit for the water-insoluble B. cenocepacia biofilm polysaccharide: [3)-α-d-Galp-(1→3)-α-d-Glcp-(1→3)-α-d-Galp-(1→3)-α-d-Manp-(1→]n Molecular modelling was used, coupled with NMR Nuclear Overhauser Effect (NOE) data, to obtain information about local structural motifs which could give hints about the polysaccharide insolubility. Both modelling and NMR data pointed at restricted dynamics of local conformations which were ascribed to the presence of inter-residue hydrogen bonds and to steric restrictions. In addition, the good correlation between NOE data and calculated interatomic distances by molecular dynamics simulations validated potential energy functions used for calculations. View Full-Text
Keywords: Burkholderia cenocepacia H111; biofilm exopolysaccharides; NMR; molecular modelling; polysaccharide structure Burkholderia cenocepacia H111; biofilm exopolysaccharides; NMR; molecular modelling; polysaccharide structure
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MDPI and ACS Style

Bellich, B.; Jou, I.A.; Caterino, M.; Rizzo, R.; Ravenscroft, N.; Fazli, M.; Tolker-Nielsen, T.; Brady, J.W.; Cescutti, P. Burkholderia cenocepacia H111 Produces a Water-Insoluble Exopolysaccharide in Biofilm: Structural Determination and Molecular Modelling. Int. J. Mol. Sci. 2020, 21, 1702. https://doi.org/10.3390/ijms21051702

AMA Style

Bellich B, Jou IA, Caterino M, Rizzo R, Ravenscroft N, Fazli M, Tolker-Nielsen T, Brady JW, Cescutti P. Burkholderia cenocepacia H111 Produces a Water-Insoluble Exopolysaccharide in Biofilm: Structural Determination and Molecular Modelling. International Journal of Molecular Sciences. 2020; 21(5):1702. https://doi.org/10.3390/ijms21051702

Chicago/Turabian Style

Bellich, Barbara; Jou, Ining A.; Caterino, Marco; Rizzo, Roberto; Ravenscroft, Neil; Fazli, Mustafa; Tolker-Nielsen, Tim; Brady, John W.; Cescutti, Paola. 2020. "Burkholderia cenocepacia H111 Produces a Water-Insoluble Exopolysaccharide in Biofilm: Structural Determination and Molecular Modelling" Int. J. Mol. Sci. 21, no. 5: 1702. https://doi.org/10.3390/ijms21051702

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