Heterologous expression of a thermophilic diacylglycerol acyltransferase triggers triglyceride accumulation in Escherichia coli
Autor(a) principal: | |
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Data de Publicação: | 2017 |
Outros Autores: | , , , , , |
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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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 |
dc.format.none.fl_str_mv |
application/pdf |
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) |
repository.mail.fl_str_mv |
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1808129340959883264 |