Cell wall modifications that alter the exolytic activity of lactococcal phage endolysins have little impact on phage growth

Bacteriophages are a nuisance in the production of fermented dairy products driven by starter bacteria and strategies to reduce the risk of phage infection are permanently sought. Bearing in mind that the bacterial cell wall plays a pivotal role in host recognition and lysis, our goal was to elucida...

Descripción completa

Detalles Bibliográficos
Autores: Escobedo, Susana, Pérez de Pipaon, Mikel, Rendueles Martínez, Claudia, Rodríguez González, Ana, Martínez Fernández, Beatriz
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2023
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/287277
Acceso en línea:http://hdl.handle.net/10261/287277
Access Level:acceso abierto
Palabra clave:Cell envelope stress
Bacteriophage
Endolysins
Lactococcus
Peptidoglycan
id ES_ed8c809e4ba3cd435f4f49bf3a67deb2
oai_identifier_str oai:digital.csic.es:10261/287277
network_acronym_str ES
network_name_str España
repository_id_str
spelling Cell wall modifications that alter the exolytic activity of lactococcal phage endolysins have little impact on phage growthEscobedo, SusanaPérez de Pipaon, MikelRendueles Martínez, ClaudiaRodríguez González, AnaMartínez Fernández, BeatrizCell envelope stressBacteriophageEndolysinsLactococcusPeptidoglycanBacteriophages are a nuisance in the production of fermented dairy products driven by starter bacteria and strategies to reduce the risk of phage infection are permanently sought. Bearing in mind that the bacterial cell wall plays a pivotal role in host recognition and lysis, our goal was to elucidate to which extent modifications in the cell wall may alter endolysin activity and influence the outcome of phage infection in Lactococcus. Three lactococcal endolysins with distinct catalytic domains (CHAP, amidase and lysozyme) from phages 1,358, p2 and c2 respectively, were purified and their exolytic activity was tested against lactococcal mutants either overexpressing or lacking genes involved in the cell envelope stress (CES) response or in modifying peptidoglycan (PG) composition. After recombinant production in E. coli, Lys1358 (CHAP) and LysC2 (muramidase) were able to lyse lactococcal cells in turbidity reduction assays, but no activity of LysP2 was detected. The degree of PG acetylation, namely C6-O-acetylation and de-N-acetylation influenced the exolytic activity, being LysC2 more active against cells depleted of the PG deacetylase PgdA and the O-acetyl transferase OatA. On the contrary, both endolysins showed reduced activity on cells with an induced CES response. By measuring several growth parameters of phage c2 on these lactococcal mutants (lytic score, efficiency of plaquing, plaque size and one-step curves), a direct link between the exolytic activity of its endolysin and phage performance could not be stablished.This work has been funded by grants AYUD/2021/52120 (Program of Science, Technology and Innovation 2018–2022, Principado de Asturias, FICYT, FEDER-UE), grant BIO2017-88147-R (MCIN/AEI/10.13039/501100011033 and by “ERDF A way of making Europe”) and grant PID2020-119697RB-I00 (MCIN/AEI/10.13039/501100011033). CR is a fellow of the program “Ayudas Severo Ochoa” of the Principality of Asturias (BP20 006).Peer reviewedFrontiers MediaPrincipado de AsturiasEuropean CommissionAgencia Estatal de Investigación (España)Ministerio de Ciencia e Innovación (España)Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]202320232023info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionapplication/pdfhttp://hdl.handle.net/10261/287277reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglés#PLACEHOLDER_PARENT_METADATA_VALUE##PLACEHOLDER_PARENT_METADATA_VALUE#info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/BIO2017-88147-Rinfo:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/PID2020-119697RB-I00Escobedo, Susana; Pérez de Pipaon, Mikel; Rendueles Martínez, Claudia; Rodríguez González, Ana; Martínez Fernández, Beatriz; 2022; Effect of cell wall modifications on exolytic activity of lactococcal phage endolysins and phage growth [Dataset]; DIGITAL.CSIC; https://doi.org/10.20350/digitalCSIC/14766https://doi.org/10.3389/fmicb.2023.1106049Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/2872772026-05-22T06:33:51Z
dc.title.none.fl_str_mv Cell wall modifications that alter the exolytic activity of lactococcal phage endolysins have little impact on phage growth
title Cell wall modifications that alter the exolytic activity of lactococcal phage endolysins have little impact on phage growth
spellingShingle Cell wall modifications that alter the exolytic activity of lactococcal phage endolysins have little impact on phage growth
Escobedo, Susana
Cell envelope stress
Bacteriophage
Endolysins
Lactococcus
Peptidoglycan
title_short Cell wall modifications that alter the exolytic activity of lactococcal phage endolysins have little impact on phage growth
title_full Cell wall modifications that alter the exolytic activity of lactococcal phage endolysins have little impact on phage growth
title_fullStr Cell wall modifications that alter the exolytic activity of lactococcal phage endolysins have little impact on phage growth
title_full_unstemmed Cell wall modifications that alter the exolytic activity of lactococcal phage endolysins have little impact on phage growth
title_sort Cell wall modifications that alter the exolytic activity of lactococcal phage endolysins have little impact on phage growth
dc.creator.none.fl_str_mv Escobedo, Susana
Pérez de Pipaon, Mikel
Rendueles Martínez, Claudia
Rodríguez González, Ana
Martínez Fernández, Beatriz
author Escobedo, Susana
author_facet Escobedo, Susana
Pérez de Pipaon, Mikel
Rendueles Martínez, Claudia
Rodríguez González, Ana
Martínez Fernández, Beatriz
author_role author
author2 Pérez de Pipaon, Mikel
Rendueles Martínez, Claudia
Rodríguez González, Ana
Martínez Fernández, Beatriz
author2_role author
author
author
author
dc.contributor.none.fl_str_mv Principado de Asturias
European Commission
Agencia Estatal de Investigación (España)
Ministerio de Ciencia e Innovación (España)
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv Cell envelope stress
Bacteriophage
Endolysins
Lactococcus
Peptidoglycan
topic Cell envelope stress
Bacteriophage
Endolysins
Lactococcus
Peptidoglycan
description Bacteriophages are a nuisance in the production of fermented dairy products driven by starter bacteria and strategies to reduce the risk of phage infection are permanently sought. Bearing in mind that the bacterial cell wall plays a pivotal role in host recognition and lysis, our goal was to elucidate to which extent modifications in the cell wall may alter endolysin activity and influence the outcome of phage infection in Lactococcus. Three lactococcal endolysins with distinct catalytic domains (CHAP, amidase and lysozyme) from phages 1,358, p2 and c2 respectively, were purified and their exolytic activity was tested against lactococcal mutants either overexpressing or lacking genes involved in the cell envelope stress (CES) response or in modifying peptidoglycan (PG) composition. After recombinant production in E. coli, Lys1358 (CHAP) and LysC2 (muramidase) were able to lyse lactococcal cells in turbidity reduction assays, but no activity of LysP2 was detected. The degree of PG acetylation, namely C6-O-acetylation and de-N-acetylation influenced the exolytic activity, being LysC2 more active against cells depleted of the PG deacetylase PgdA and the O-acetyl transferase OatA. On the contrary, both endolysins showed reduced activity on cells with an induced CES response. By measuring several growth parameters of phage c2 on these lactococcal mutants (lytic score, efficiency of plaquing, plaque size and one-step curves), a direct link between the exolytic activity of its endolysin and phage performance could not be stablished.
publishDate 2023
dc.date.none.fl_str_mv 2023
2023
2023
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
Publisher's version
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/287277
url http://hdl.handle.net/10261/287277
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv #PLACEHOLDER_PARENT_METADATA_VALUE#
#PLACEHOLDER_PARENT_METADATA_VALUE#
info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/BIO2017-88147-R
info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/PID2020-119697RB-I00
Escobedo, Susana; Pérez de Pipaon, Mikel; Rendueles Martínez, Claudia; Rodríguez González, Ana; Martínez Fernández, Beatriz; 2022; Effect of cell wall modifications on exolytic activity of lactococcal phage endolysins and phage growth [Dataset]; DIGITAL.CSIC; https://doi.org/10.20350/digitalCSIC/14766
https://doi.org/10.3389/fmicb.2023.1106049
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Frontiers Media
publisher.none.fl_str_mv Frontiers Media
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
repository.name.fl_str_mv
repository.mail.fl_str_mv
_version_ 1869423514366246912
score 15,811543

Similares en LA Referencia