A methodology for the selection and characterization of riboflavin-overproducing Weissella cibaria strains after treatment with roseoflavin
Fermentative processes by lactic acid bacteria can produce metabolites of interest to the health and food industries. Two examples are the production of B-group vitamins, and of prebiotic and immunomodulatory dextran-type exopolysaccharides. In this study, three riboflavin- and dextran-producing Wei...
| Autores: | , , , , , , , |
|---|---|
| Tipo de recurso: | artículo |
| Fecha de publicación: | 2023 |
| País: | España |
| Institución: | Universidad del País Vasco |
| Repositorio: | Addi. Archivo Digital para la Docencia y la Investigación |
| OAI Identifier: | oai:addi.ehu.eus:10810/61899 |
| Acceso en línea: | http://hdl.handle.net/10810/61899 |
| Access Level: | acceso abierto |
| Palabra clave: | Weissella cibaria lactic acid bacteria vitamin B2 (riboflavin) FMN riboswitch roseoflavin riboflavin overproducing bacteria regulation of rib operon |
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| dc.title.none.fl_str_mv |
A methodology for the selection and characterization of riboflavin-overproducing Weissella cibaria strains after treatment with roseoflavin |
| title |
A methodology for the selection and characterization of riboflavin-overproducing Weissella cibaria strains after treatment with roseoflavin |
| spellingShingle |
A methodology for the selection and characterization of riboflavin-overproducing Weissella cibaria strains after treatment with roseoflavin Díez Ozaeta, Iñaki Weissella cibaria lactic acid bacteria vitamin B2 (riboflavin) FMN riboswitch roseoflavin riboflavin overproducing bacteria regulation of rib operon |
| title_short |
A methodology for the selection and characterization of riboflavin-overproducing Weissella cibaria strains after treatment with roseoflavin |
| title_full |
A methodology for the selection and characterization of riboflavin-overproducing Weissella cibaria strains after treatment with roseoflavin |
| title_fullStr |
A methodology for the selection and characterization of riboflavin-overproducing Weissella cibaria strains after treatment with roseoflavin |
| title_full_unstemmed |
A methodology for the selection and characterization of riboflavin-overproducing Weissella cibaria strains after treatment with roseoflavin |
| title_sort |
A methodology for the selection and characterization of riboflavin-overproducing Weissella cibaria strains after treatment with roseoflavin |
| dc.creator.none.fl_str_mv |
Díez Ozaeta, Iñaki Martín Loarte, Lucía Mohedano, Mari Luz Tamame, Mercedes Ruiz Masó, José Ángel Del Solar, Gloria Dueñas Chasco, María Teresa López García, Paloma |
| author |
Díez Ozaeta, Iñaki |
| author_facet |
Díez Ozaeta, Iñaki Martín Loarte, Lucía Mohedano, Mari Luz Tamame, Mercedes Ruiz Masó, José Ángel Del Solar, Gloria Dueñas Chasco, María Teresa López García, Paloma |
| author_role |
author |
| author2 |
Martín Loarte, Lucía Mohedano, Mari Luz Tamame, Mercedes Ruiz Masó, José Ángel Del Solar, Gloria Dueñas Chasco, María Teresa López García, Paloma |
| author2_role |
author author author author author author author |
| dc.subject.none.fl_str_mv |
Weissella cibaria lactic acid bacteria vitamin B2 (riboflavin) FMN riboswitch roseoflavin riboflavin overproducing bacteria regulation of rib operon |
| topic |
Weissella cibaria lactic acid bacteria vitamin B2 (riboflavin) FMN riboswitch roseoflavin riboflavin overproducing bacteria regulation of rib operon |
| description |
Fermentative processes by lactic acid bacteria can produce metabolites of interest to the health and food industries. Two examples are the production of B-group vitamins, and of prebiotic and immunomodulatory dextran-type exopolysaccharides. In this study, three riboflavin- and dextran-producing Weissella cibaria strains (BAL3C-5, BAL3C-7 and BAL3C-22) were used to develop a new method for selection and isolation of spontaneous riboflavin-overproducing W. cibaria mutants. This method was based on the selection of strains resistant to roseoflavin. The DNA sequencing of the FMN riboswitch of bacterial cell populations treated with various roseoflavin concentrations, revealed the existence of at least 10 spontaneous and random point mutations at this location. Folding and analysis of the mutated FMN riboswitches with the RNA fold program predicted that these mutations could result in a deregulation of the rib operon expression. When the roseoflavin-treated cultures were plated on medium supporting dextran synthesis, the most promising mutants were identified by the yellow color of their mucous colonies, exhibiting a ropy phenotype. After their isolation and recovery in liquid medium, the evaluation of their riboflavin production revealed that the mutant strains synthesized a wide range of riboflavin levels (from 0.80 to 6.50 mg/L) above the wild-type level (0.15 mg/L). Thus, this was a reliable method to select spontaneous riboflavin-overproducing and dextran-producing strains of W. cibaria. This species has not yet been used as a starter or adjunct culture, but this study reinforces the potential that it has for the food and health industry for the production of functional foods or as a probiotic. Furthermore, analysis of the influence of FMN present in the growth medium, on rib mRNA and riboflavin levels, revealed which mutant strains produce riboflavin without flavin regulation. Moreover, the BAL3C-5 C120T mutant was identified as the highest riboflavin-overproducer. Determination of its chromosomal DNA sequence and that of BAL3C-5, revealed a total identity between the 2 strains except for the C120T mutation at the FMN riboswitch. To our knowledge, this work is the first demonstration that only a single alteration in the genome of a lactic acid bacteria is required for a riboflavin-overproducing phenotype. |
| publishDate |
2023 |
| dc.date.none.fl_str_mv |
2023 2023 2023 |
| dc.type.none.fl_str_mv |
info:eu-repo/semantics/article |
| format |
article |
| dc.identifier.none.fl_str_mv |
http://hdl.handle.net/10810/61899 |
| url |
http://hdl.handle.net/10810/61899 |
| dc.language.none.fl_str_mv |
Inglés |
| language_invalid_str_mv |
Inglés |
| dc.relation.none.fl_str_mv |
https://www.frontiersin.org/articles/10.3389/fmicb.2023.1154130/full |
| dc.rights.none.fl_str_mv |
info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/3.0/es/ Atribución 3.0 España |
| eu_rights_str_mv |
openAccess |
| rights_invalid_str_mv |
http://creativecommons.org/licenses/by/3.0/es/ Atribución 3.0 España |
| dc.format.none.fl_str_mv |
application/pdf |
| dc.publisher.none.fl_str_mv |
Frontiers Media |
| publisher.none.fl_str_mv |
Frontiers Media |
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reponame:Addi. Archivo Digital para la Docencia y la Investigación instname:Universidad del País Vasco |
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Universidad del País Vasco |
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Addi. Archivo Digital para la Docencia y la Investigación |
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Addi. Archivo Digital para la Docencia y la Investigación |
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1869411856938958848 |
| spelling |
A methodology for the selection and characterization of riboflavin-overproducing Weissella cibaria strains after treatment with roseoflavinDíez Ozaeta, IñakiMartín Loarte, LucíaMohedano, Mari LuzTamame, MercedesRuiz Masó, José ÁngelDel Solar, GloriaDueñas Chasco, María TeresaLópez García, PalomaWeissella cibarialactic acid bacteriavitamin B2 (riboflavin)FMN riboswitchroseoflavinriboflavin overproducing bacteriaregulation of rib operonFermentative processes by lactic acid bacteria can produce metabolites of interest to the health and food industries. Two examples are the production of B-group vitamins, and of prebiotic and immunomodulatory dextran-type exopolysaccharides. In this study, three riboflavin- and dextran-producing Weissella cibaria strains (BAL3C-5, BAL3C-7 and BAL3C-22) were used to develop a new method for selection and isolation of spontaneous riboflavin-overproducing W. cibaria mutants. This method was based on the selection of strains resistant to roseoflavin. The DNA sequencing of the FMN riboswitch of bacterial cell populations treated with various roseoflavin concentrations, revealed the existence of at least 10 spontaneous and random point mutations at this location. Folding and analysis of the mutated FMN riboswitches with the RNA fold program predicted that these mutations could result in a deregulation of the rib operon expression. When the roseoflavin-treated cultures were plated on medium supporting dextran synthesis, the most promising mutants were identified by the yellow color of their mucous colonies, exhibiting a ropy phenotype. After their isolation and recovery in liquid medium, the evaluation of their riboflavin production revealed that the mutant strains synthesized a wide range of riboflavin levels (from 0.80 to 6.50 mg/L) above the wild-type level (0.15 mg/L). Thus, this was a reliable method to select spontaneous riboflavin-overproducing and dextran-producing strains of W. cibaria. This species has not yet been used as a starter or adjunct culture, but this study reinforces the potential that it has for the food and health industry for the production of functional foods or as a probiotic. Furthermore, analysis of the influence of FMN present in the growth medium, on rib mRNA and riboflavin levels, revealed which mutant strains produce riboflavin without flavin regulation. Moreover, the BAL3C-5 C120T mutant was identified as the highest riboflavin-overproducer. Determination of its chromosomal DNA sequence and that of BAL3C-5, revealed a total identity between the 2 strains except for the C120T mutation at the FMN riboswitch. To our knowledge, this work is the first demonstration that only a single alteration in the genome of a lactic acid bacteria is required for a riboflavin-overproducing phenotype.We acknowledge support of the publication fee by the CSIC Open Access Publication Support Initiative through its Unit of Information Resources for Research (URICI). In addition, this work is also based upon the work from COST Action 18101 SOURDOMICS – Sourdough biotechnology network towards novel, healthier and sustainable food and bioprocesses (https://sourdomics.com/; https://www.cost.eu/actions/CA18101/#tabs|Name:overview), where the authors PL, GdS, ML-M, JR-M, MT, and MT are Members of the Working Groups WG2 and WG3.Frontiers Media202320232023info:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10810/61899reponame:Addi. Archivo Digital para la Docencia y la Investigacióninstname:Universidad del País VascoIngléshttps://www.frontiersin.org/articles/10.3389/fmicb.2023.1154130/fullinfo:eu-repo/semantics/openAccesshttp://creativecommons.org/licenses/by/3.0/es/© 2023 Diez-Ozaeta, Martín-Loarte, Mohedano, Tamame, Ruiz-Masó, del Solar, Dueñas and López. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.Atribución 3.0 Españaoai:addi.ehu.eus:10810/618992026-06-18T09:23:17Z |
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15,300719 |