Effect of a mutagenized acyl-ACP thioesterase FATA allele from sunflower with improved activity in tobacco leaves and Arabidopsis seeds

The substrate specificity of the acyl-acyl carrier protein (ACP) thioesterases significantly determines the type of fatty acids that are exported from plastids. Thus, designing acyl-ACP thioesterases with different substrate specificities or kinetic properties would be of interest for plant lipid bi...

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Autores: Moreno-Pérez, Antonio J., Venegas-Calerón, Mónica, Vaistij, Fabián E., Salas, Joaquín J., Larson, Tony R., Garcés Mancheño, Rafael, Graham, Ian A., Martínez-Force, Enrique
Tipo de recurso: artículo
Fecha de publicación:2014
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/115002
Acceso en línea:http://hdl.handle.net/10261/115002
Access Level:acceso abierto
Palabra clave:Triacylglycerols
Transient expression
Tobacco
Helianthus annuus
Fatty acids
Arabidopsis thaliana
Acyl-ACP thioesterases
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spelling Effect of a mutagenized acyl-ACP thioesterase FATA allele from sunflower with improved activity in tobacco leaves and Arabidopsis seedsMoreno-Pérez, Antonio J.Venegas-Calerón, MónicaVaistij, Fabián E.Salas, Joaquín J.Larson, Tony R.Garcés Mancheño, RafaelGraham, Ian A.Martínez-Force, EnriqueTriacylglycerolsTransient expressionTobaccoHelianthus annuusFatty acidsArabidopsis thalianaAcyl-ACP thioesterasesThe substrate specificity of the acyl-acyl carrier protein (ACP) thioesterases significantly determines the type of fatty acids that are exported from plastids. Thus, designing acyl-ACP thioesterases with different substrate specificities or kinetic properties would be of interest for plant lipid biotechnology to produce oils enriched in specialty fatty acids. In the present work, the FatA thioesterase from Helianthus annuus was used to test the impact of changes in the amino acids present in the binding pocket on substrate specificity and catalytic efficiency. Amongst all the mutated enzymes studied, Q215W was especially interesting as it had higher specificity towards saturated acyl-ACP substrates and higher catalytic efficiency compared to wild-type H. annuus FatA. Null, wild type and high-efficiency alleles were transiently expressed in tobacco leaves to check their effect on lipid biosynthesis. Expression of active FatA thioesterases altered the composition of leaf triacylglycerols but did not alter total lipid content. However, the expression of the wild type and the high-efficiency alleles in Arabidopsis thaliana transgenic seeds resulted in a strong reduction in oil content and an increase in total saturated fatty acid content. The role and influence of acyl-ACP thioesterases in plant metabolism and their possible applications in lipid biotechnology are discussed.This work was supported by the MINECO and FEDER (Project AGL2011-23187)Peer ReviewedSpringer Nature2015201520142015info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501http://hdl.handle.net/10261/115002reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglésinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/1150022026-05-22T06:33:51Z
dc.title.none.fl_str_mv Effect of a mutagenized acyl-ACP thioesterase FATA allele from sunflower with improved activity in tobacco leaves and Arabidopsis seeds
title Effect of a mutagenized acyl-ACP thioesterase FATA allele from sunflower with improved activity in tobacco leaves and Arabidopsis seeds
spellingShingle Effect of a mutagenized acyl-ACP thioesterase FATA allele from sunflower with improved activity in tobacco leaves and Arabidopsis seeds
Moreno-Pérez, Antonio J.
Triacylglycerols
Transient expression
Tobacco
Helianthus annuus
Fatty acids
Arabidopsis thaliana
Acyl-ACP thioesterases
title_short Effect of a mutagenized acyl-ACP thioesterase FATA allele from sunflower with improved activity in tobacco leaves and Arabidopsis seeds
title_full Effect of a mutagenized acyl-ACP thioesterase FATA allele from sunflower with improved activity in tobacco leaves and Arabidopsis seeds
title_fullStr Effect of a mutagenized acyl-ACP thioesterase FATA allele from sunflower with improved activity in tobacco leaves and Arabidopsis seeds
title_full_unstemmed Effect of a mutagenized acyl-ACP thioesterase FATA allele from sunflower with improved activity in tobacco leaves and Arabidopsis seeds
title_sort Effect of a mutagenized acyl-ACP thioesterase FATA allele from sunflower with improved activity in tobacco leaves and Arabidopsis seeds
dc.creator.none.fl_str_mv Moreno-Pérez, Antonio J.
Venegas-Calerón, Mónica
Vaistij, Fabián E.
Salas, Joaquín J.
Larson, Tony R.
Garcés Mancheño, Rafael
Graham, Ian A.
Martínez-Force, Enrique
author Moreno-Pérez, Antonio J.
author_facet Moreno-Pérez, Antonio J.
Venegas-Calerón, Mónica
Vaistij, Fabián E.
Salas, Joaquín J.
Larson, Tony R.
Garcés Mancheño, Rafael
Graham, Ian A.
Martínez-Force, Enrique
author_role author
author2 Venegas-Calerón, Mónica
Vaistij, Fabián E.
Salas, Joaquín J.
Larson, Tony R.
Garcés Mancheño, Rafael
Graham, Ian A.
Martínez-Force, Enrique
author2_role author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Triacylglycerols
Transient expression
Tobacco
Helianthus annuus
Fatty acids
Arabidopsis thaliana
Acyl-ACP thioesterases
topic Triacylglycerols
Transient expression
Tobacco
Helianthus annuus
Fatty acids
Arabidopsis thaliana
Acyl-ACP thioesterases
description The substrate specificity of the acyl-acyl carrier protein (ACP) thioesterases significantly determines the type of fatty acids that are exported from plastids. Thus, designing acyl-ACP thioesterases with different substrate specificities or kinetic properties would be of interest for plant lipid biotechnology to produce oils enriched in specialty fatty acids. In the present work, the FatA thioesterase from Helianthus annuus was used to test the impact of changes in the amino acids present in the binding pocket on substrate specificity and catalytic efficiency. Amongst all the mutated enzymes studied, Q215W was especially interesting as it had higher specificity towards saturated acyl-ACP substrates and higher catalytic efficiency compared to wild-type H. annuus FatA. Null, wild type and high-efficiency alleles were transiently expressed in tobacco leaves to check their effect on lipid biosynthesis. Expression of active FatA thioesterases altered the composition of leaf triacylglycerols but did not alter total lipid content. However, the expression of the wild type and the high-efficiency alleles in Arabidopsis thaliana transgenic seeds resulted in a strong reduction in oil content and an increase in total saturated fatty acid content. The role and influence of acyl-ACP thioesterases in plant metabolism and their possible applications in lipid biotechnology are discussed.
publishDate 2014
dc.date.none.fl_str_mv 2014
2015
2015
2015
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
format article
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/115002
url http://hdl.handle.net/10261/115002
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Springer Nature
publisher.none.fl_str_mv Springer Nature
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
repository.name.fl_str_mv
repository.mail.fl_str_mv
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