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Liposomal Delivery of Newly Identified Prophage Lysins in a Pseudomonas aeruginosa Model

2022-09-04Journal article Open access
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dc.contributor.authorMorais, Diana
dc.contributor.authorTanoeiro, Luís
dc.contributor.authorMarques, Andreia
dc.contributor.authorGonçalves, Tiago
dc.contributor.authorDuarte, A.
dc.contributor.authorMatos, António
dc.contributor.authorVital, Joana
dc.contributor.authorCruz, Maria Eugénia Meirinhos
dc.contributor.authorCarvalheiro, Manuela Colla
dc.contributor.authorAnes, Elsa
dc.contributor.authorVítor, Jorge M. B.
dc.contributor.authorGaspar, Maria Manuela
dc.contributor.authorVale, Filipa F.
dc.date.accessioned2025-08-14T17:43:54Z
dc.date.available2025-08-14T17:43:54Z
dc.date.issued2022-09-04
dc.date.updated2024-01-25T17:19:01Z
dc.description© 2022 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).pt_PT
dc.description.abstractPseudomonas aeruginosa is a Gram-negative opportunistic bacterium that presents resistance to several antibiotics, thus, representing a major threat to human and animal health. Phage-derived products, namely lysins, or peptidoglycan-hydrolyzing enzymes, can be an effective weapon against antibiotic-resistant bacteria. Whereas in Gram-positive bacteria, lysis from without is facilitated by the exposed peptidoglycan layer, this is not possible in the outer membrane-protected peptidoglycan of Gram-negative bacteria. Here, we suggest the encapsulation of lysins in liposomes as a delivery system against Gram-negative bacteria, using the model of P. aeruginosa. Bioinformatic analysis allowed for the identification of 38 distinct complete prophages within 66 P. aeruginosa genomes (16 of which newly sequenced) and led to the identification of 19 lysins of diverse sequence and function, 5 of which proceeded to wet lab analysis. The four purifiable lysins showed hydrolytic activity against Gram-positive bacterial lawns and, on zymogram assays, constituted of autoclaved P. aeruginosa cells. Additionally, lysins Pa7 and Pa119 combined with an outer membrane permeabilizer showed activity against P. aeruginosa cells. These two lysins were successfully encapsulated in DMPC:DOPE:CHEMS (molar ratio 4:4:2) liposomes with an average encapsulation efficiency of 33.33% and 32.30%, respectively. The application of the encapsulated lysins to the model P. aeruginosa led to a reduction in cell viability and resulted in cell lysis as observed in MTT cell viability assays and electron microscopy. In sum, we report here that prophages may be important sources of new enzybiotics, with prophage lysins showing high diversity and activity. In addition, these enzybiotics following their incorporation in liposomes were able to potentiate their antibacterial effect against the Gram-negative bacteria P. aeruginosa, used as the model.pt_PT
dc.description.sponsorshipFFV is funded by Fundação para a Ciência e a Tecnologia (FCT) through an Assistant Researcher grant CEECIND/03023/2017, and a project grant (project PTDC/BTM-SAL/28978/2017) that supported this work. JSV holds a research fellowship within the scope of project PTDC/BTM-TEC/3238/2020 (FCT). The work has been partially supported by National funds from FCT, projects UIDB/04138/2020 and UIDP/04138/2020.pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.doi10.3390/ijms231710143pt_PT
dc.identifier.issn1422-0067
dc.identifier.slugcv-prod-3158721
dc.identifier.urihttp://hdl.handle.net/10400.5/102919
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherMDPIpt_PT
dc.relationNot Available
dc.relationPhage-Enzybiotic: dealing with critical pathogenic antibiotic-resistant bacteria
dc.relationIn evolution and disease: the underlying role of Helicobacter prophages
dc.relationResearch Institute for Medicines
dc.relationResearch Institute for Medicines
dc.relation.publisherversionhttps://www.mdpi.com/1422-0067/23/17/10143pt_PT
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/pt_PT
dc.subjectPseudomonas aeruginosapt_PT
dc.subjectProphagept_PT
dc.subjectBacteriophagept_PT
dc.subjectPhage therapypt_PT
dc.subjectLysinspt_PT
dc.subjectAntibiotic resistancept_PT
dc.subjectLiposomespt_PT
dc.subjectGram-negative bacteriapt_PT
dc.titleLiposomal Delivery of Newly Identified Prophage Lysins in a Pseudomonas aeruginosa Modelpt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.awardTitleNot Available
oaire.awardTitlePhage-Enzybiotic: dealing with critical pathogenic antibiotic-resistant bacteria
oaire.awardTitleIn evolution and disease: the underlying role of Helicobacter prophages
oaire.awardTitleResearch Institute for Medicines
oaire.awardTitleResearch Institute for Medicines
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/CEEC IND 2017/CEECIND%2F03023%2F2017%2FCP1476%2FCT0004/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/3599-PPCDT/PTDC%2FBTM-SAL%2F28978%2F2017/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/Concurso para Financiamento de Projetos de Investigação Científica e Desenvolvimento Tecnológico em Todos os Domínios Científicos - 2020/PTDC%2FBTM-TEC%2F3238%2F2020/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDB%2F04138%2F2020/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDP%2F04138%2F2020/PT
oaire.citation.issue17pt_PT
oaire.citation.startPage10143pt_PT
oaire.citation.titleInternational Journal of Molecular Sciencespt_PT
oaire.citation.volume23pt_PT
oaire.fundingStreamCEEC IND 2017
oaire.fundingStream3599-PPCDT
oaire.fundingStreamConcurso para Financiamento de Projetos de Investigação Científica e Desenvolvimento Tecnológico em Todos os Domínios Científicos - 2020
oaire.fundingStream6817 - DCRRNI ID
oaire.fundingStream6817 - DCRRNI ID
person.familyNameDuarte
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person.familyNameVítor
person.familyNamede Jesus Guilherme Gaspar
person.familyNameVale
person.givenNameAida
person.givenNameManuela
person.givenNameELSA MARIA
person.givenNameJorge
person.givenNameMaria Manuela
person.givenNameFilipa
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person.identifier.orcid0000-0003-4635-0105
person.identifier.ridM-5272-2013
person.identifier.ridM-3279-2013
person.identifier.ridC-3570-2013
person.identifier.scopus-author-id35495748300
person.identifier.scopus-author-id57202998389
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project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.nameFundação para a Ciência e a Tecnologia
project.funder.nameFundação para a Ciência e a Tecnologia
project.funder.nameFundação para a Ciência e a Tecnologia
project.funder.nameFundação para a Ciência e a Tecnologia
project.funder.nameFundação para a Ciência e a Tecnologia
rcaap.cv.cienciaid4718-00EF-4BF0 | Ana Filipa Ferreira do Vale
rcaap.rightsopenAccesspt_PT
rcaap.typearticlept_PT
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