Characterization of different alginate lyases for dissolving Pseudomonas aeruginosa biofilms

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  • dc.contributor.author Blanco Cabra, Núria
  • dc.contributor.author Paetzold, Bernhard, 1981-
  • dc.contributor.author Ferrar, Tony
  • dc.contributor.author Mazzolini, Rocco
  • dc.contributor.author Torrents, Eduard
  • dc.contributor.author Serrano Pubull, Luis, 1982-
  • dc.contributor.author Lluch-Senar, Maria 1982-
  • dc.date.accessioned 2020-10-16T06:34:07Z
  • dc.date.available 2020-10-16T06:34:07Z
  • dc.date.issued 2020
  • dc.description.abstract Aggregates of Pseudomonas aeruginosa form a protective barrier against antibiotics and the immune system. These barriers, known as biofilms, are associated with several infectious diseases. One of the main components of these biofilms is alginate, a homo- and hetero-polysaccharide that consists of β-D-mannuronate (M) and α-L-guluronate (G) units. Alginate lyases degrade this sugar and have been proposed as biotherapeutic agents to dissolve P. aeruginosa biofilms. However, there are contradictory reports in the literature regarding the efficacy of alginate lyases against biofilms and their synergistic effect with antibiotics. We found that most positive reports used a commercial crude extract from Flavobacterium multivorum as the alginate lyase source. By using anion exchange chromatography coupled to nano LC MS/MS, we identified two distinct enzymes in this extract, one has both polyM and polyG (polyM/G) degradation activities and it is similar in sequence to a broad-spectrum alginate lyase from Flavobacterium sp. S20 (Alg2A). The other enzyme has only polyG activity and it is similar in sequence to AlyA1 from Zobellia galactanivorans. By characterizing both of these enzymes together with three recombinant alginate lyases (a polyM, a polyG and a polyM/G), we showed that only enzymes with polyM/G activity such as Alg2A and A1-II' (alginate lyase from Sphingomonas sp.) are effective in dissolving biofilms. Furthermore, both activities are required to have a synergistic effect with antibiotics.
  • dc.description.sponsorship We acknowledge support of the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation program under agreement No 670216 (MYCOCHASSIS), the Spanish Ministry of Economy and Competitiveness and Fondo Europeo de Desarrollo Regional (MINECO-FEDER) (BIO2015-63557-R), ‘Centro de Excelencia Severo Ochoa 2013-2017’, FEDER project from Instituto Carlos III (ISCIII, Acción Estratégica en Salud 2016) (reference CP16/00094) and “Secretaria d’Universitats i Recerca del Departament d’Economia i Coneixement de la Generalitat de Catalunya / CERCA programme” (2014SGR678 and 2017SGR1079). The CRG/UPF Proteomics Unit is part of the “Plataforma de Recursos Biomoleculares y Bioinformáticos (ProteoRed)” supported by grant PT13/0001 of Instituto de Salud Carlos III from the Spanish Government.
  • dc.format.mimetype application/pdf
  • dc.identifier.citation Blanco-Cabra N, Paetzold B, Ferrar T, Mazzolini R, Torrents E, Serrano L, LLuch-Senar M. Characterization of different alginate lyases for dissolving Pseudomonas aeruginosa biofilms. Sci Rep. 2020; 10(1):9390. DOI: 10.1038/s41598-020-66293-2
  • dc.identifier.doi http://dx.doi.org/10.1038/s41598-020-66293-2
  • dc.identifier.issn 2045-2322
  • dc.identifier.uri http://hdl.handle.net/10230/45498
  • dc.language.iso eng
  • dc.publisher Nature Research
  • dc.relation.ispartof Sci Rep. 2020; 10(1):9390
  • dc.relation.projectID info:eu-repo/grantAgreement/EC/H2020/670216
  • dc.relation.projectID info:eu-repo/grantAgreement/ES/1PE/BIO2015-63557-R
  • dc.rights © The Author(s) 2020. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
  • dc.rights.accessRights info:eu-repo/semantics/openAccess
  • dc.rights.uri http://creativecommons.org/licenses/by/4.0/
  • dc.subject.keyword Applied microbiology
  • dc.subject.keyword Biofilms
  • dc.title Characterization of different alginate lyases for dissolving Pseudomonas aeruginosa biofilms
  • dc.type info:eu-repo/semantics/article
  • dc.type.version info:eu-repo/semantics/publishedVersion

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