Heterologous expression of a thermophilic diacylglycerol acyltransferase triggers triglyceride accumulation in Escherichia coli

Detalhes bibliográficos
Autor(a) principal: Lázaro, Beatriz [UNESP]
Data de Publicação: 2017
Outros Autores: Villa, Juan A., Santín, Omar, Cabezas, Matilde, Milagre, Cintia D.F. [UNESP], De La Cruz, Fernando, Moncalián, Gabriel
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da UNESP
Texto Completo: http://dx.doi.org/10.1371/journal.pone.0176520
http://hdl.handle.net/11449/174500
Resumo: Triglycerides (TAGs), the major storage molecules of metabolic energy and source of fatty acids, are produced as single cell oil by some oleogenic microorganisms. However, these microorganisms require strict culture conditions, show low carbon source flexibilities, lack efficient genetic modification tools and in some cases pose safety concerns. TAGs have essential applications such as behaving as a source for added-value fatty acids or giving rise to the production of biodiesel. Hence, new alternative methods are urgently required for obtaining these oils. In this work we describe TAG accumulation in the industrially appropriate microorganism Escherichia coli expressing the heterologous enzyme tDGAT, a wax ester synthase/triacylglycerol:acylCoA acyltranferase (WS/DGAT). With this purpose, we introduce a codon-optimized gene from the thermophilic actinomycete Thermomonospora curvata coding for a WS/DGAT into different E. coli strains, describe the metabolic effects associated to the expression of this protein and evaluate neutral lipid accumulation. We observe a direct relation between the expression of this WS/DGAT and TAG production within a wide range of culture conditions. More than 30% TAGs were detected within the bacterial neutral lipids in 90 minutes after induction. TAGs were observed to be associated with the hydrophobic enzyme while forming round intracytoplasmic bodies, which could represent a bottleneck for lipid accumulation in E. coli. We detected an increase of almost 3- fold in the monounsaturated fatty acids (MUFA) occurring in the recombinant strains. These MUFA were predominant in the accumulated TAGs achieving 46% of the TAG fatty acids. These results set the basis for further research on the achievement of a suitable method towards the sustainable production of these neutral lipids.
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spelling Heterologous expression of a thermophilic diacylglycerol acyltransferase triggers triglyceride accumulation in Escherichia coliTriglycerides (TAGs), the major storage molecules of metabolic energy and source of fatty acids, are produced as single cell oil by some oleogenic microorganisms. However, these microorganisms require strict culture conditions, show low carbon source flexibilities, lack efficient genetic modification tools and in some cases pose safety concerns. TAGs have essential applications such as behaving as a source for added-value fatty acids or giving rise to the production of biodiesel. Hence, new alternative methods are urgently required for obtaining these oils. In this work we describe TAG accumulation in the industrially appropriate microorganism Escherichia coli expressing the heterologous enzyme tDGAT, a wax ester synthase/triacylglycerol:acylCoA acyltranferase (WS/DGAT). With this purpose, we introduce a codon-optimized gene from the thermophilic actinomycete Thermomonospora curvata coding for a WS/DGAT into different E. coli strains, describe the metabolic effects associated to the expression of this protein and evaluate neutral lipid accumulation. We observe a direct relation between the expression of this WS/DGAT and TAG production within a wide range of culture conditions. More than 30% TAGs were detected within the bacterial neutral lipids in 90 minutes after induction. TAGs were observed to be associated with the hydrophobic enzyme while forming round intracytoplasmic bodies, which could represent a bottleneck for lipid accumulation in E. coli. We detected an increase of almost 3- fold in the monounsaturated fatty acids (MUFA) occurring in the recombinant strains. These MUFA were predominant in the accumulated TAGs achieving 46% of the TAG fatty acids. These results set the basis for further research on the achievement of a suitable method towards the sustainable production of these neutral lipids.European CommissionDepartamento de Biología Molecular Universidad de Cantabria Instituto de Biomedicina y Biotecnología de Cantabria (IBBTEC) Consejo Superior de Investigaciones Científicas - Universidad de Cantabria, C/Albert EinsteinDepartment of Organic Chemistry Institute of Chemistry Universidade Estadual Paulista (UNESP), Rua Prof. Francisco DegniDepartment of Molecular Microbiology and Immunology Saint Louis University Liver Center Saint Louis University School of MedicineDepartment of Organic Chemistry Institute of Chemistry Universidade Estadual Paulista (UNESP), Rua Prof. Francisco DegniEuropean Commission: IB14DM0176Consejo Superior de Investigaciones Científicas - Universidad de CantabriaUniversidade Estadual Paulista (Unesp)Saint Louis University School of MedicineLázaro, Beatriz [UNESP]Villa, Juan A.Santín, OmarCabezas, MatildeMilagre, Cintia D.F. [UNESP]De La Cruz, FernandoMoncalián, Gabriel2018-12-11T17:11:26Z2018-12-11T17:11:26Z2017-04-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://dx.doi.org/10.1371/journal.pone.0176520PLoS ONE, v. 12, n. 4, 2017.1932-6203http://hdl.handle.net/11449/17450010.1371/journal.pone.01765202-s2.0-850183449472-s2.0-85018344947.pdf14257489168493760000-0001-5627-8616Scopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengPLoS ONE1,164info:eu-repo/semantics/openAccess2023-12-29T06:20:41Zoai:repositorio.unesp.br:11449/174500Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T21:36:39.938417Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false
dc.title.none.fl_str_mv Heterologous expression of a thermophilic diacylglycerol acyltransferase triggers triglyceride accumulation in Escherichia coli
title Heterologous expression of a thermophilic diacylglycerol acyltransferase triggers triglyceride accumulation in Escherichia coli
spellingShingle Heterologous expression of a thermophilic diacylglycerol acyltransferase triggers triglyceride accumulation in Escherichia coli
Lázaro, Beatriz [UNESP]
title_short Heterologous expression of a thermophilic diacylglycerol acyltransferase triggers triglyceride accumulation in Escherichia coli
title_full Heterologous expression of a thermophilic diacylglycerol acyltransferase triggers triglyceride accumulation in Escherichia coli
title_fullStr Heterologous expression of a thermophilic diacylglycerol acyltransferase triggers triglyceride accumulation in Escherichia coli
title_full_unstemmed Heterologous expression of a thermophilic diacylglycerol acyltransferase triggers triglyceride accumulation in Escherichia coli
title_sort Heterologous expression of a thermophilic diacylglycerol acyltransferase triggers triglyceride accumulation in Escherichia coli
author Lázaro, Beatriz [UNESP]
author_facet Lázaro, Beatriz [UNESP]
Villa, Juan A.
Santín, Omar
Cabezas, Matilde
Milagre, Cintia D.F. [UNESP]
De La Cruz, Fernando
Moncalián, Gabriel
author_role author
author2 Villa, Juan A.
Santín, Omar
Cabezas, Matilde
Milagre, Cintia D.F. [UNESP]
De La Cruz, Fernando
Moncalián, Gabriel
author2_role author
author
author
author
author
author
dc.contributor.none.fl_str_mv Consejo Superior de Investigaciones Científicas - Universidad de Cantabria
Universidade Estadual Paulista (Unesp)
Saint Louis University School of Medicine
dc.contributor.author.fl_str_mv Lázaro, Beatriz [UNESP]
Villa, Juan A.
Santín, Omar
Cabezas, Matilde
Milagre, Cintia D.F. [UNESP]
De La Cruz, Fernando
Moncalián, Gabriel
description Triglycerides (TAGs), the major storage molecules of metabolic energy and source of fatty acids, are produced as single cell oil by some oleogenic microorganisms. However, these microorganisms require strict culture conditions, show low carbon source flexibilities, lack efficient genetic modification tools and in some cases pose safety concerns. TAGs have essential applications such as behaving as a source for added-value fatty acids or giving rise to the production of biodiesel. Hence, new alternative methods are urgently required for obtaining these oils. In this work we describe TAG accumulation in the industrially appropriate microorganism Escherichia coli expressing the heterologous enzyme tDGAT, a wax ester synthase/triacylglycerol:acylCoA acyltranferase (WS/DGAT). With this purpose, we introduce a codon-optimized gene from the thermophilic actinomycete Thermomonospora curvata coding for a WS/DGAT into different E. coli strains, describe the metabolic effects associated to the expression of this protein and evaluate neutral lipid accumulation. We observe a direct relation between the expression of this WS/DGAT and TAG production within a wide range of culture conditions. More than 30% TAGs were detected within the bacterial neutral lipids in 90 minutes after induction. TAGs were observed to be associated with the hydrophobic enzyme while forming round intracytoplasmic bodies, which could represent a bottleneck for lipid accumulation in E. coli. We detected an increase of almost 3- fold in the monounsaturated fatty acids (MUFA) occurring in the recombinant strains. These MUFA were predominant in the accumulated TAGs achieving 46% of the TAG fatty acids. These results set the basis for further research on the achievement of a suitable method towards the sustainable production of these neutral lipids.
publishDate 2017
dc.date.none.fl_str_mv 2017-04-01
2018-12-11T17:11:26Z
2018-12-11T17:11:26Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://dx.doi.org/10.1371/journal.pone.0176520
PLoS ONE, v. 12, n. 4, 2017.
1932-6203
http://hdl.handle.net/11449/174500
10.1371/journal.pone.0176520
2-s2.0-85018344947
2-s2.0-85018344947.pdf
1425748916849376
0000-0001-5627-8616
url http://dx.doi.org/10.1371/journal.pone.0176520
http://hdl.handle.net/11449/174500
identifier_str_mv PLoS ONE, v. 12, n. 4, 2017.
1932-6203
10.1371/journal.pone.0176520
2-s2.0-85018344947
2-s2.0-85018344947.pdf
1425748916849376
0000-0001-5627-8616
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv PLoS ONE
1,164
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
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dc.source.none.fl_str_mv Scopus
reponame:Repositório Institucional da UNESP
instname:Universidade Estadual Paulista (UNESP)
instacron:UNESP
instname_str Universidade Estadual Paulista (UNESP)
instacron_str UNESP
institution UNESP
reponame_str Repositório Institucional da UNESP
collection Repositório Institucional da UNESP
repository.name.fl_str_mv Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)
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