Understanding the interplay of carbon and nitrogen supply for ectoines production and metabolic overflow in high density cultures of Chromohalobacter salexigens

Background The halophilic bacterium Chromohalobacter salexigens has been proposed as promising cell factory for the production of the compatible solutes ectoine and hydroxyectoine. This bacterium has evolved metabolic adaptations to efficiently grow under high salt concentrations by accumulating ect...

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Autores: Salar García, María J., Bernal, Vicente, Pastor, José M., Salvador de Lara, Manuel, Argandoña Bertrán, Montserrat, Nieto Gutiérrez, Joaquín José, Vargas Macías, Carmen
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2017
País:España
Institución:Universidad de Sevilla (US)
Repositorio:idUS. Depósito de Investigación de la Universidad de Sevilla
OAI Identifier:oai:idus.us.es:11441/59966
Acceso en línea:http://hdl.handle.net/11441/59966
https://doi.org/10.1186/s12934-017-0643-7
Access Level:acceso abierto
Palabra clave:C. salexigens
Ectoines
Carbon overflow
Halophilism
Fed-batch
Nitrogen assimilation
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spelling Understanding the interplay of carbon and nitrogen supply for ectoines production and metabolic overflow in high density cultures of Chromohalobacter salexigensSalar García, María J.Bernal, VicentePastor, José M.Salvador de Lara, ManuelArgandoña Bertrán, MontserratNieto Gutiérrez, Joaquín JoséVargas Macías, CarmenC. salexigensEctoinesCarbon overflowHalophilismFed-batchNitrogen assimilationBackground The halophilic bacterium Chromohalobacter salexigens has been proposed as promising cell factory for the production of the compatible solutes ectoine and hydroxyectoine. This bacterium has evolved metabolic adaptations to efficiently grow under high salt concentrations by accumulating ectoines as compatible solutes. However, metabolic overflow, which is a major drawback for the efficient conversion of biological feedstocks, occurs as a result of metabolic unbalances during growth and ectoines production. Optimal production of ectoines is conditioned by the interplay of carbon and nitrogen metabolisms. In this work, we set out to determine how nitrogen supply affects the production of ectoines. Results Chromohalobacter salexigens was challenged to grow in media with unbalanced carbon/nitrogen ratio. In C. salexigens, overflow metabolism and ectoines production are a function of medium composition. At low ammonium conditions, the growth rate decreased importantly, up to 80%. Shifts in overflow metabolism were observed when changing the C/N ratio in the culture medium. 13C-NMR analysis of ectoines labelling revealed a high metabolic rigidity, with almost constant flux ratios in all conditions assayed. Unbalanced C/N ratio led to pyruvate accumulation, especially upon N-limitation. Analysis of an ect − mutant demonstrated the link between metabolic overflow and ectoine biosynthesis. Under non ectoine synthesizing conditions, glucose uptake and metabolic overflow decreased importantly. Finally, in fed-batch cultures, biomass yield was affected by the feeding scheme chosen. High growth (up to 42.4 g L−1) and volumetric ectoine yields (up to 4.21 g L−1) were obtained by minimizing metabolite overflow and nutrient accumulation in high density cultures in a low nitrogen fed-batch culture. Moreover, the yield coefficient calculated for the transformation of glucose into biomass was 30% higher in fed-batch than in the batch culture, demonstrating that the metabolic efficiency of C. salexigens can be improved by careful design of culture feeding schemes. Conclusions Metabolic shifts observed at low ammonium concentrations were explained by a shift in the energy required for nitrogen assimilation. Carbon-limited fed-batch cultures with reduced ammonium supply were the best conditions for cultivation of C. salexigens, supporting high density growth and maintaining high ectoines production.España, Ministerio de Ciencia e Innovación BIO2011-29233-C02-01España, Ministerio de Economía y Competitividad BIO2014-54411-C2-1-RBioMed CentralMicrobiología y ParasitologíaMinisterio de Ciencia e Innovación (MICIN). EspañaMinisterio de Economía y Competitividad (MINECO). España2017info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfapplication/pdfhttp://hdl.handle.net/11441/59966https://doi.org/10.1186/s12934-017-0643-7reponame:idUS. Depósito de Investigación de la Universidad de Sevillainstname:Universidad de Sevilla (US)InglésMicrobial Cell Factories, 16 (1), 1-12.BIO2011-29233-C02-01info:eu-repo/grantAgreement/MINECO/BIO2014-54411-C2-1-R/http://dx.doi.org/10.1186/s12934-017-0643-7.info:eu-repo/semantics/openAccessoai:idus.us.es:11441/599662026-06-17T12:51:07Z
dc.title.none.fl_str_mv Understanding the interplay of carbon and nitrogen supply for ectoines production and metabolic overflow in high density cultures of Chromohalobacter salexigens
title Understanding the interplay of carbon and nitrogen supply for ectoines production and metabolic overflow in high density cultures of Chromohalobacter salexigens
spellingShingle Understanding the interplay of carbon and nitrogen supply for ectoines production and metabolic overflow in high density cultures of Chromohalobacter salexigens
Salar García, María J.
C. salexigens
Ectoines
Carbon overflow
Halophilism
Fed-batch
Nitrogen assimilation
title_short Understanding the interplay of carbon and nitrogen supply for ectoines production and metabolic overflow in high density cultures of Chromohalobacter salexigens
title_full Understanding the interplay of carbon and nitrogen supply for ectoines production and metabolic overflow in high density cultures of Chromohalobacter salexigens
title_fullStr Understanding the interplay of carbon and nitrogen supply for ectoines production and metabolic overflow in high density cultures of Chromohalobacter salexigens
title_full_unstemmed Understanding the interplay of carbon and nitrogen supply for ectoines production and metabolic overflow in high density cultures of Chromohalobacter salexigens
title_sort Understanding the interplay of carbon and nitrogen supply for ectoines production and metabolic overflow in high density cultures of Chromohalobacter salexigens
dc.creator.none.fl_str_mv Salar García, María J.
Bernal, Vicente
Pastor, José M.
Salvador de Lara, Manuel
Argandoña Bertrán, Montserrat
Nieto Gutiérrez, Joaquín José
Vargas Macías, Carmen
author Salar García, María J.
author_facet Salar García, María J.
Bernal, Vicente
Pastor, José M.
Salvador de Lara, Manuel
Argandoña Bertrán, Montserrat
Nieto Gutiérrez, Joaquín José
Vargas Macías, Carmen
author_role author
author2 Bernal, Vicente
Pastor, José M.
Salvador de Lara, Manuel
Argandoña Bertrán, Montserrat
Nieto Gutiérrez, Joaquín José
Vargas Macías, Carmen
author2_role author
author
author
author
author
author
dc.contributor.none.fl_str_mv Microbiología y Parasitología
Ministerio de Ciencia e Innovación (MICIN). España
Ministerio de Economía y Competitividad (MINECO). España
dc.subject.none.fl_str_mv C. salexigens
Ectoines
Carbon overflow
Halophilism
Fed-batch
Nitrogen assimilation
topic C. salexigens
Ectoines
Carbon overflow
Halophilism
Fed-batch
Nitrogen assimilation
description Background The halophilic bacterium Chromohalobacter salexigens has been proposed as promising cell factory for the production of the compatible solutes ectoine and hydroxyectoine. This bacterium has evolved metabolic adaptations to efficiently grow under high salt concentrations by accumulating ectoines as compatible solutes. However, metabolic overflow, which is a major drawback for the efficient conversion of biological feedstocks, occurs as a result of metabolic unbalances during growth and ectoines production. Optimal production of ectoines is conditioned by the interplay of carbon and nitrogen metabolisms. In this work, we set out to determine how nitrogen supply affects the production of ectoines. Results Chromohalobacter salexigens was challenged to grow in media with unbalanced carbon/nitrogen ratio. In C. salexigens, overflow metabolism and ectoines production are a function of medium composition. At low ammonium conditions, the growth rate decreased importantly, up to 80%. Shifts in overflow metabolism were observed when changing the C/N ratio in the culture medium. 13C-NMR analysis of ectoines labelling revealed a high metabolic rigidity, with almost constant flux ratios in all conditions assayed. Unbalanced C/N ratio led to pyruvate accumulation, especially upon N-limitation. Analysis of an ect − mutant demonstrated the link between metabolic overflow and ectoine biosynthesis. Under non ectoine synthesizing conditions, glucose uptake and metabolic overflow decreased importantly. Finally, in fed-batch cultures, biomass yield was affected by the feeding scheme chosen. High growth (up to 42.4 g L−1) and volumetric ectoine yields (up to 4.21 g L−1) were obtained by minimizing metabolite overflow and nutrient accumulation in high density cultures in a low nitrogen fed-batch culture. Moreover, the yield coefficient calculated for the transformation of glucose into biomass was 30% higher in fed-batch than in the batch culture, demonstrating that the metabolic efficiency of C. salexigens can be improved by careful design of culture feeding schemes. Conclusions Metabolic shifts observed at low ammonium concentrations were explained by a shift in the energy required for nitrogen assimilation. Carbon-limited fed-batch cultures with reduced ammonium supply were the best conditions for cultivation of C. salexigens, supporting high density growth and maintaining high ectoines production.
publishDate 2017
dc.date.none.fl_str_mv 2017
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11441/59966
https://doi.org/10.1186/s12934-017-0643-7
url http://hdl.handle.net/11441/59966
https://doi.org/10.1186/s12934-017-0643-7
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv Microbial Cell Factories, 16 (1), 1-12.
BIO2011-29233-C02-01
info:eu-repo/grantAgreement/MINECO/BIO2014-54411-C2-1-R/
http://dx.doi.org/10.1186/s12934-017-0643-7.
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv BioMed Central
publisher.none.fl_str_mv BioMed Central
dc.source.none.fl_str_mv reponame:idUS. Depósito de Investigación de la Universidad de Sevilla
instname:Universidad de Sevilla (US)
instname_str Universidad de Sevilla (US)
reponame_str idUS. Depósito de Investigación de la Universidad de Sevilla
collection idUS. Depósito de Investigación de la Universidad de Sevilla
repository.name.fl_str_mv
repository.mail.fl_str_mv
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