Characterization of different alginate lyases for dissolving Pseudomonas aeruginosa biofilms

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...

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Autores: Blanco Cabra, Núria, Paetzold, Bernhard, 1981-, Ferrar, Tony, Mazzolini, Rocco, Torrents, Eduard, Serrano Pubull, Luis, 1982-, Lluch-Senar, Maria 1982-
Formato: artículo
Estado:Versión publicada
Fecha de publicación:2020
País:España
Recursos:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
Repositorio:Recercat. Dipósit de la Recerca de Catalunya
OAI Identifier:oai:recercat.cat:10230/45498
Acesso em linha:http://hdl.handle.net/10230/45498
http://dx.doi.org/10.1038/s41598-020-66293-2
Access Level:acceso abierto
Palavra-chave:Applied microbiology
Biofilms
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dc.title.none.fl_str_mv Characterization of different alginate lyases for dissolving Pseudomonas aeruginosa biofilms
title Characterization of different alginate lyases for dissolving Pseudomonas aeruginosa biofilms
spellingShingle Characterization of different alginate lyases for dissolving Pseudomonas aeruginosa biofilms
Blanco Cabra, Núria
Applied microbiology
Biofilms
title_short Characterization of different alginate lyases for dissolving Pseudomonas aeruginosa biofilms
title_full Characterization of different alginate lyases for dissolving Pseudomonas aeruginosa biofilms
title_fullStr Characterization of different alginate lyases for dissolving Pseudomonas aeruginosa biofilms
title_full_unstemmed Characterization of different alginate lyases for dissolving Pseudomonas aeruginosa biofilms
title_sort Characterization of different alginate lyases for dissolving Pseudomonas aeruginosa biofilms
dc.creator.none.fl_str_mv Blanco Cabra, Núria
Paetzold, Bernhard, 1981-
Ferrar, Tony
Mazzolini, Rocco
Torrents, Eduard
Serrano Pubull, Luis, 1982-
Lluch-Senar, Maria 1982-
author Blanco Cabra, Núria
author_facet Blanco Cabra, Núria
Paetzold, Bernhard, 1981-
Ferrar, Tony
Mazzolini, Rocco
Torrents, Eduard
Serrano Pubull, Luis, 1982-
Lluch-Senar, Maria 1982-
author_role author
author2 Paetzold, Bernhard, 1981-
Ferrar, Tony
Mazzolini, Rocco
Torrents, Eduard
Serrano Pubull, Luis, 1982-
Lluch-Senar, Maria 1982-
author2_role author
author
author
author
author
author
dc.subject.none.fl_str_mv Applied microbiology
Biofilms
topic Applied microbiology
Biofilms
description 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.
publishDate 2020
dc.date.none.fl_str_mv 2020
2020
2020
dc.type.none.fl_str_mv info:eu-repo/semantics/article
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dc.identifier.none.fl_str_mv http://hdl.handle.net/10230/45498
http://dx.doi.org/10.1038/s41598-020-66293-2
url http://hdl.handle.net/10230/45498
http://dx.doi.org/10.1038/s41598-020-66293-2
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv Sci Rep. 2020; 10(1):9390
info:eu-repo/grantAgreement/EC/H2020/670216
info:eu-repo/grantAgreement/ES/1PE/BIO2015-63557-R
dc.rights.none.fl_str_mv http://creativecommons.org/licenses/by/4.0/
info:eu-repo/semantics/openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by/4.0/
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dc.publisher.none.fl_str_mv Nature Research
publisher.none.fl_str_mv Nature Research
dc.source.none.fl_str_mv reponame:Recercat. Dipósit de la Recerca de Catalunya
instname:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
instname_str Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
reponame_str Recercat. Dipósit de la Recerca de Catalunya
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spelling Characterization of different alginate lyases for dissolving Pseudomonas aeruginosa biofilmsBlanco Cabra, NúriaPaetzold, Bernhard, 1981-Ferrar, TonyMazzolini, RoccoTorrents, EduardSerrano Pubull, Luis, 1982-Lluch-Senar, Maria 1982-Applied microbiologyBiofilmsAggregates 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.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.Nature Research202020202020info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfapplication/pdfhttp://hdl.handle.net/10230/45498http://dx.doi.org/10.1038/s41598-020-66293-2reponame:Recercat. Dipósit de la Recerca de Catalunyainstname:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)InglésSci Rep. 2020; 10(1):9390info:eu-repo/grantAgreement/EC/H2020/670216info:eu-repo/grantAgreement/ES/1PE/BIO2015-63557-R© 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/.http://creativecommons.org/licenses/by/4.0/info:eu-repo/semantics/openAccessoai:recercat.cat:10230/454982026-05-29T05:05:01Z
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