Characterization of the sorbitol utilization cluster of the probiotic Pediococcus parvulus 2.6: Genetic, functional and complementation studies in heterologous hosts

Autores
Pérez Ramos, Adrian; Werning, María Laura; Prieto, Alicia; Russo, Pasquale; Spano, Giuseppe; Mohedano, Mari L.; López, Paloma
Año de publicación
2017
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Pediococcus parvulus 2.6 secretes a 2-substituted (1,3)-β-D-glucan with prebiotic and immunomodulatory properties. It is synthesized by the GTF glycosyltransferase using UDP-glucose as substrate. Analysis of the P. parvulus 2.6 draft genome revealed the existence of a sorbitol utilization cluster of six genes (gutFRMCBA), whose products should be involved in sorbitol utilization and could generate substrates for UDP-glucose synthesis. Southern blot hybridization analysis showed that the cluster is located in a plasmid. Analysis of metabolic fluxes and production of the exopolysaccharide revealed that: (i) P. parvulus 2.6 is able to metabolize sorbitol, (ii) sorbitol utilization is repressed in the presence of glucose and (iii) sorbitol supports the synthesis of 2-substituted (1,3)-β-D-glucan. The sorbitol cluster encodes two putative regulators, GutR and GutM, in addition to a phosphoenolpyruvate-dependent phosphotransferase transport system and sorbitol-6-phosphate dehydrogenase. Therefore, we investigated the involvement of GutR and GutM in the expression of gutFRMCBA. The promoter-probe vector pRCR based on the mrfp gene, which encodes the fluorescence protein mCherry, was used to test the potential promoter of the cluster (Pgut) and the genes encoding the regulators. This was performed by transferring by electrotransformation the recombinant plasmids into two hosts, which metabolize sorbitol: Lactobacillus plantarum and Lactobacillus casei. Upon growth in the presence of sorbitol, but not of glucose, only the presence of Pgut was required to support expression of mrfp in L. plantarum. In L. casei the presence of sorbitol in the growth medium and the pediococcal gutR or gutR plus gutM in the genome was required for Pgut functionality. This demonstrates that: (i) Pgut is required for expression of the gut cluster, (ii) Pgut is subjected to catabolic repression in lactobacilli, (iii) GutR is an activator, and (iv) in the presence of sorbitol, trans-complementation for activation of Pgut exists in L. plantarum but not in L. casei.
Fil: Pérez Ramos, Adrian. Consejo Superior de Investigaciones Científicas; España
Fil: Werning, María Laura. Consejo Superior de Investigaciones Científicas; España. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia de Catamarca. Universidad Nacional de Catamarca. Centro de Investigaciones y Transferencia de Catamarca; Argentina
Fil: Prieto, Alicia. Consejo Superior de Investigaciones Científicas; España
Fil: Russo, Pasquale. University of Foggia; Italia
Fil: Spano, Giuseppe. University of Foggia; Italia
Fil: Mohedano, Mari L.. Consejo Superior de Investigaciones Científicas; España
Fil: López, Paloma. Consejo Superior de Investigaciones Científicas; España
Materia
EXOPOLYSACCHARIDES
LACTIC ACID BACTERIA
PEDIOCOCCUS PARVULUS
PROBIOTIC
SORBITOL
Β-GLUCANS
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/41495
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/41495
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Characterization of the sorbitol utilization cluster of the probiotic Pediococcus parvulus 2.6: Genetic, functional and complementation studies in heterologous hostsPérez Ramos, AdrianWerning, María LauraPrieto, AliciaRusso, PasqualeSpano, GiuseppeMohedano, Mari L.López, PalomaEXOPOLYSACCHARIDESLACTIC ACID BACTERIAPEDIOCOCCUS PARVULUSPROBIOTICSORBITOLΒ-GLUCANShttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Pediococcus parvulus 2.6 secretes a 2-substituted (1,3)-β-D-glucan with prebiotic and immunomodulatory properties. It is synthesized by the GTF glycosyltransferase using UDP-glucose as substrate. Analysis of the P. parvulus 2.6 draft genome revealed the existence of a sorbitol utilization cluster of six genes (gutFRMCBA), whose products should be involved in sorbitol utilization and could generate substrates for UDP-glucose synthesis. Southern blot hybridization analysis showed that the cluster is located in a plasmid. Analysis of metabolic fluxes and production of the exopolysaccharide revealed that: (i) P. parvulus 2.6 is able to metabolize sorbitol, (ii) sorbitol utilization is repressed in the presence of glucose and (iii) sorbitol supports the synthesis of 2-substituted (1,3)-β-D-glucan. The sorbitol cluster encodes two putative regulators, GutR and GutM, in addition to a phosphoenolpyruvate-dependent phosphotransferase transport system and sorbitol-6-phosphate dehydrogenase. Therefore, we investigated the involvement of GutR and GutM in the expression of gutFRMCBA. The promoter-probe vector pRCR based on the mrfp gene, which encodes the fluorescence protein mCherry, was used to test the potential promoter of the cluster (Pgut) and the genes encoding the regulators. This was performed by transferring by electrotransformation the recombinant plasmids into two hosts, which metabolize sorbitol: Lactobacillus plantarum and Lactobacillus casei. Upon growth in the presence of sorbitol, but not of glucose, only the presence of Pgut was required to support expression of mrfp in L. plantarum. In L. casei the presence of sorbitol in the growth medium and the pediococcal gutR or gutR plus gutM in the genome was required for Pgut functionality. This demonstrates that: (i) Pgut is required for expression of the gut cluster, (ii) Pgut is subjected to catabolic repression in lactobacilli, (iii) GutR is an activator, and (iv) in the presence of sorbitol, trans-complementation for activation of Pgut exists in L. plantarum but not in L. casei.Fil: Pérez Ramos, Adrian. Consejo Superior de Investigaciones Científicas; EspañaFil: Werning, María Laura. Consejo Superior de Investigaciones Científicas; España. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia de Catamarca. Universidad Nacional de Catamarca. Centro de Investigaciones y Transferencia de Catamarca; ArgentinaFil: Prieto, Alicia. Consejo Superior de Investigaciones Científicas; EspañaFil: Russo, Pasquale. University of Foggia; ItaliaFil: Spano, Giuseppe. University of Foggia; ItaliaFil: Mohedano, Mari L.. Consejo Superior de Investigaciones Científicas; EspañaFil: López, Paloma. Consejo Superior de Investigaciones Científicas; EspañaFrontiers Media S.A.2017-12info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/41495Pérez Ramos, Adrian; Werning, María Laura; Prieto, Alicia; Russo, Pasquale; Spano, Giuseppe; et al.; Characterization of the sorbitol utilization cluster of the probiotic Pediococcus parvulus 2.6: Genetic, functional and complementation studies in heterologous hosts; Frontiers Media S.A.; Frontiers in Microbiology; 8; 12-2017; 1-171664-302XCONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.3389/fmicb.2017.02393info:eu-repo/semantics/altIdentifier/url/https://www.frontiersin.org/articles/10.3389/fmicb.2017.02393/fullinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-22T11:33:45Zoai:ri.conicet.gov.ar:11336/41495instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-10-22 11:33:45.708CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Characterization of the sorbitol utilization cluster of the probiotic Pediococcus parvulus 2.6: Genetic, functional and complementation studies in heterologous hosts
title Characterization of the sorbitol utilization cluster of the probiotic Pediococcus parvulus 2.6: Genetic, functional and complementation studies in heterologous hosts
spellingShingle Characterization of the sorbitol utilization cluster of the probiotic Pediococcus parvulus 2.6: Genetic, functional and complementation studies in heterologous hosts
Pérez Ramos, Adrian
EXOPOLYSACCHARIDES
LACTIC ACID BACTERIA
PEDIOCOCCUS PARVULUS
PROBIOTIC
SORBITOL
Β-GLUCANS
title_short Characterization of the sorbitol utilization cluster of the probiotic Pediococcus parvulus 2.6: Genetic, functional and complementation studies in heterologous hosts
title_full Characterization of the sorbitol utilization cluster of the probiotic Pediococcus parvulus 2.6: Genetic, functional and complementation studies in heterologous hosts
title_fullStr Characterization of the sorbitol utilization cluster of the probiotic Pediococcus parvulus 2.6: Genetic, functional and complementation studies in heterologous hosts
title_full_unstemmed Characterization of the sorbitol utilization cluster of the probiotic Pediococcus parvulus 2.6: Genetic, functional and complementation studies in heterologous hosts
title_sort Characterization of the sorbitol utilization cluster of the probiotic Pediococcus parvulus 2.6: Genetic, functional and complementation studies in heterologous hosts
dc.creator.none.fl_str_mv Pérez Ramos, Adrian
Werning, María Laura
Prieto, Alicia
Russo, Pasquale
Spano, Giuseppe
Mohedano, Mari L.
López, Paloma
author Pérez Ramos, Adrian
author_facet Pérez Ramos, Adrian
Werning, María Laura
Prieto, Alicia
Russo, Pasquale
Spano, Giuseppe
Mohedano, Mari L.
López, Paloma
author_role author
author2 Werning, María Laura
Prieto, Alicia
Russo, Pasquale
Spano, Giuseppe
Mohedano, Mari L.
López, Paloma
author2_role author
author
author
author
author
author
dc.subject.none.fl_str_mv EXOPOLYSACCHARIDES
LACTIC ACID BACTERIA
PEDIOCOCCUS PARVULUS
PROBIOTIC
SORBITOL
Β-GLUCANS
topic EXOPOLYSACCHARIDES
LACTIC ACID BACTERIA
PEDIOCOCCUS PARVULUS
PROBIOTIC
SORBITOL
Β-GLUCANS
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Pediococcus parvulus 2.6 secretes a 2-substituted (1,3)-β-D-glucan with prebiotic and immunomodulatory properties. It is synthesized by the GTF glycosyltransferase using UDP-glucose as substrate. Analysis of the P. parvulus 2.6 draft genome revealed the existence of a sorbitol utilization cluster of six genes (gutFRMCBA), whose products should be involved in sorbitol utilization and could generate substrates for UDP-glucose synthesis. Southern blot hybridization analysis showed that the cluster is located in a plasmid. Analysis of metabolic fluxes and production of the exopolysaccharide revealed that: (i) P. parvulus 2.6 is able to metabolize sorbitol, (ii) sorbitol utilization is repressed in the presence of glucose and (iii) sorbitol supports the synthesis of 2-substituted (1,3)-β-D-glucan. The sorbitol cluster encodes two putative regulators, GutR and GutM, in addition to a phosphoenolpyruvate-dependent phosphotransferase transport system and sorbitol-6-phosphate dehydrogenase. Therefore, we investigated the involvement of GutR and GutM in the expression of gutFRMCBA. The promoter-probe vector pRCR based on the mrfp gene, which encodes the fluorescence protein mCherry, was used to test the potential promoter of the cluster (Pgut) and the genes encoding the regulators. This was performed by transferring by electrotransformation the recombinant plasmids into two hosts, which metabolize sorbitol: Lactobacillus plantarum and Lactobacillus casei. Upon growth in the presence of sorbitol, but not of glucose, only the presence of Pgut was required to support expression of mrfp in L. plantarum. In L. casei the presence of sorbitol in the growth medium and the pediococcal gutR or gutR plus gutM in the genome was required for Pgut functionality. This demonstrates that: (i) Pgut is required for expression of the gut cluster, (ii) Pgut is subjected to catabolic repression in lactobacilli, (iii) GutR is an activator, and (iv) in the presence of sorbitol, trans-complementation for activation of Pgut exists in L. plantarum but not in L. casei.
Fil: Pérez Ramos, Adrian. Consejo Superior de Investigaciones Científicas; España
Fil: Werning, María Laura. Consejo Superior de Investigaciones Científicas; España. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia de Catamarca. Universidad Nacional de Catamarca. Centro de Investigaciones y Transferencia de Catamarca; Argentina
Fil: Prieto, Alicia. Consejo Superior de Investigaciones Científicas; España
Fil: Russo, Pasquale. University of Foggia; Italia
Fil: Spano, Giuseppe. University of Foggia; Italia
Fil: Mohedano, Mari L.. Consejo Superior de Investigaciones Científicas; España
Fil: López, Paloma. Consejo Superior de Investigaciones Científicas; España
description Pediococcus parvulus 2.6 secretes a 2-substituted (1,3)-β-D-glucan with prebiotic and immunomodulatory properties. It is synthesized by the GTF glycosyltransferase using UDP-glucose as substrate. Analysis of the P. parvulus 2.6 draft genome revealed the existence of a sorbitol utilization cluster of six genes (gutFRMCBA), whose products should be involved in sorbitol utilization and could generate substrates for UDP-glucose synthesis. Southern blot hybridization analysis showed that the cluster is located in a plasmid. Analysis of metabolic fluxes and production of the exopolysaccharide revealed that: (i) P. parvulus 2.6 is able to metabolize sorbitol, (ii) sorbitol utilization is repressed in the presence of glucose and (iii) sorbitol supports the synthesis of 2-substituted (1,3)-β-D-glucan. The sorbitol cluster encodes two putative regulators, GutR and GutM, in addition to a phosphoenolpyruvate-dependent phosphotransferase transport system and sorbitol-6-phosphate dehydrogenase. Therefore, we investigated the involvement of GutR and GutM in the expression of gutFRMCBA. The promoter-probe vector pRCR based on the mrfp gene, which encodes the fluorescence protein mCherry, was used to test the potential promoter of the cluster (Pgut) and the genes encoding the regulators. This was performed by transferring by electrotransformation the recombinant plasmids into two hosts, which metabolize sorbitol: Lactobacillus plantarum and Lactobacillus casei. Upon growth in the presence of sorbitol, but not of glucose, only the presence of Pgut was required to support expression of mrfp in L. plantarum. In L. casei the presence of sorbitol in the growth medium and the pediococcal gutR or gutR plus gutM in the genome was required for Pgut functionality. This demonstrates that: (i) Pgut is required for expression of the gut cluster, (ii) Pgut is subjected to catabolic repression in lactobacilli, (iii) GutR is an activator, and (iv) in the presence of sorbitol, trans-complementation for activation of Pgut exists in L. plantarum but not in L. casei.
publishDate 2017
dc.date.none.fl_str_mv 2017-12
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/41495
Pérez Ramos, Adrian; Werning, María Laura; Prieto, Alicia; Russo, Pasquale; Spano, Giuseppe; et al.; Characterization of the sorbitol utilization cluster of the probiotic Pediococcus parvulus 2.6: Genetic, functional and complementation studies in heterologous hosts; Frontiers Media S.A.; Frontiers in Microbiology; 8; 12-2017; 1-17
1664-302X
CONICET Digital
CONICET
url http://hdl.handle.net/11336/41495
identifier_str_mv Pérez Ramos, Adrian; Werning, María Laura; Prieto, Alicia; Russo, Pasquale; Spano, Giuseppe; et al.; Characterization of the sorbitol utilization cluster of the probiotic Pediococcus parvulus 2.6: Genetic, functional and complementation studies in heterologous hosts; Frontiers Media S.A.; Frontiers in Microbiology; 8; 12-2017; 1-17
1664-302X
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.3389/fmicb.2017.02393
info:eu-repo/semantics/altIdentifier/url/https://www.frontiersin.org/articles/10.3389/fmicb.2017.02393/full
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Frontiers Media S.A.
publisher.none.fl_str_mv Frontiers Media S.A.
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 12.982451

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