ATP-Independent and Cell-Free Biosynthesis of β-Hydroxy Acids Using Vinyl Esters as Smart Substrates

Autores
Orrego, Alejandro H.; Grazia Rubanu, Maria; López, Idania L.; Andrés Sanz, Daniel; García-Marquina, Guillermo; Pieslinger, German Eduardo; Salassa, Luca; López Gallego, Fernando
Año de publicación
2023
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
In vitro biosynthetic pathways that condense and reduce molecules through coenzyme A (CoASH) activation demand energy and redox power in the form of ATP and NAD(P)H, respectively. These coenzymes must be orthogonally recycled by ancillary reactions that consume chemicals, electricity, or light, impacting the atom economy and/or the energy consumption of the biosystem. In this work, we have exploited vinyl esters as dual acyl and electron donor substrates to synthesize β-hydroxy acids through a non-decarboxylating Claisen condensation, reduction and hydrolysis stepwise cascade, including a NADH recycling step, catalyzed by a total of 4 enzymes. Herein, the chemical energy to activate the acyl group with CoASH and the redox power for the reduction are embedded into the vinyl esters. Upon optimization, this self-sustaining cascade reached a titer of (S)-3-hydroxy butyrate of 24 mM without requiring ATP and simultaneously recycling CoASH and NADH. This work illustrates the potential of in vitro biocatalysis to transform simple molecules into multi-functional ones.
Fil: Orrego, Alejandro H.. No especifíca;
Fil: Grazia Rubanu, Maria. No especifíca;
Fil: López, Idania L.. No especifíca;
Fil: Andrés Sanz, Daniel. No especifíca;
Fil: García-Marquina, Guillermo. No especifíca;
Fil: Pieslinger, German Eduardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina
Fil: Salassa, Luca. Universidad del País Vasco; España
Fil: López Gallego, Fernando. Universidad del País Vasco; España
Materia
CELL-FREE BIOCATALYSIS
COENZYME A
DEHYDROGENASES
MULTI-ENZYME SYSTEMS
THIOLASES
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/227763
Acceder
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repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling ATP-Independent and Cell-Free Biosynthesis of β-Hydroxy Acids Using Vinyl Esters as Smart SubstratesOrrego, Alejandro H.Grazia Rubanu, MariaLópez, Idania L.Andrés Sanz, DanielGarcía-Marquina, GuillermoPieslinger, German EduardoSalassa, LucaLópez Gallego, FernandoCELL-FREE BIOCATALYSISCOENZYME ADEHYDROGENASESMULTI-ENZYME SYSTEMSTHIOLASEShttps://purl.org/becyt/ford/1.4https://purl.org/becyt/ford/1https://purl.org/becyt/ford/1.4https://purl.org/becyt/ford/1In vitro biosynthetic pathways that condense and reduce molecules through coenzyme A (CoASH) activation demand energy and redox power in the form of ATP and NAD(P)H, respectively. These coenzymes must be orthogonally recycled by ancillary reactions that consume chemicals, electricity, or light, impacting the atom economy and/or the energy consumption of the biosystem. In this work, we have exploited vinyl esters as dual acyl and electron donor substrates to synthesize β-hydroxy acids through a non-decarboxylating Claisen condensation, reduction and hydrolysis stepwise cascade, including a NADH recycling step, catalyzed by a total of 4 enzymes. Herein, the chemical energy to activate the acyl group with CoASH and the redox power for the reduction are embedded into the vinyl esters. Upon optimization, this self-sustaining cascade reached a titer of (S)-3-hydroxy butyrate of 24 mM without requiring ATP and simultaneously recycling CoASH and NADH. This work illustrates the potential of in vitro biocatalysis to transform simple molecules into multi-functional ones.Fil: Orrego, Alejandro H.. No especifíca;Fil: Grazia Rubanu, Maria. No especifíca;Fil: López, Idania L.. No especifíca;Fil: Andrés Sanz, Daniel. No especifíca;Fil: García-Marquina, Guillermo. No especifíca;Fil: Pieslinger, German Eduardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; ArgentinaFil: Salassa, Luca. Universidad del País Vasco; EspañaFil: López Gallego, Fernando. Universidad del País Vasco; EspañaWiley VCH Verlag2023-03info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/227763Orrego, Alejandro H.; Grazia Rubanu, Maria; López, Idania L.; Andrés Sanz, Daniel; García-Marquina, Guillermo; et al.; ATP-Independent and Cell-Free Biosynthesis of β-Hydroxy Acids Using Vinyl Esters as Smart Substrates; Wiley VCH Verlag; Angewandte Chemie; 62; 13; 3-2023; 1-91433-7851CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1002/anie.202218312info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-nd/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-15T14:39:02Zoai:ri.conicet.gov.ar:11336/227763instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-10-15 14:39:03.123CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv ATP-Independent and Cell-Free Biosynthesis of β-Hydroxy Acids Using Vinyl Esters as Smart Substrates
title ATP-Independent and Cell-Free Biosynthesis of β-Hydroxy Acids Using Vinyl Esters as Smart Substrates
spellingShingle ATP-Independent and Cell-Free Biosynthesis of β-Hydroxy Acids Using Vinyl Esters as Smart Substrates
Orrego, Alejandro H.
CELL-FREE BIOCATALYSIS
COENZYME A
DEHYDROGENASES
MULTI-ENZYME SYSTEMS
THIOLASES
title_short ATP-Independent and Cell-Free Biosynthesis of β-Hydroxy Acids Using Vinyl Esters as Smart Substrates
title_full ATP-Independent and Cell-Free Biosynthesis of β-Hydroxy Acids Using Vinyl Esters as Smart Substrates
title_fullStr ATP-Independent and Cell-Free Biosynthesis of β-Hydroxy Acids Using Vinyl Esters as Smart Substrates
title_full_unstemmed ATP-Independent and Cell-Free Biosynthesis of β-Hydroxy Acids Using Vinyl Esters as Smart Substrates
title_sort ATP-Independent and Cell-Free Biosynthesis of β-Hydroxy Acids Using Vinyl Esters as Smart Substrates
dc.creator.none.fl_str_mv Orrego, Alejandro H.
Grazia Rubanu, Maria
López, Idania L.
Andrés Sanz, Daniel
García-Marquina, Guillermo
Pieslinger, German Eduardo
Salassa, Luca
López Gallego, Fernando
author Orrego, Alejandro H.
author_facet Orrego, Alejandro H.
Grazia Rubanu, Maria
López, Idania L.
Andrés Sanz, Daniel
García-Marquina, Guillermo
Pieslinger, German Eduardo
Salassa, Luca
López Gallego, Fernando
author_role author
author2 Grazia Rubanu, Maria
López, Idania L.
Andrés Sanz, Daniel
García-Marquina, Guillermo
Pieslinger, German Eduardo
Salassa, Luca
López Gallego, Fernando
author2_role author
author
author
author
author
author
author
dc.subject.none.fl_str_mv CELL-FREE BIOCATALYSIS
COENZYME A
DEHYDROGENASES
MULTI-ENZYME SYSTEMS
THIOLASES
topic CELL-FREE BIOCATALYSIS
COENZYME A
DEHYDROGENASES
MULTI-ENZYME SYSTEMS
THIOLASES
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.4
https://purl.org/becyt/ford/1
https://purl.org/becyt/ford/1.4
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv In vitro biosynthetic pathways that condense and reduce molecules through coenzyme A (CoASH) activation demand energy and redox power in the form of ATP and NAD(P)H, respectively. These coenzymes must be orthogonally recycled by ancillary reactions that consume chemicals, electricity, or light, impacting the atom economy and/or the energy consumption of the biosystem. In this work, we have exploited vinyl esters as dual acyl and electron donor substrates to synthesize β-hydroxy acids through a non-decarboxylating Claisen condensation, reduction and hydrolysis stepwise cascade, including a NADH recycling step, catalyzed by a total of 4 enzymes. Herein, the chemical energy to activate the acyl group with CoASH and the redox power for the reduction are embedded into the vinyl esters. Upon optimization, this self-sustaining cascade reached a titer of (S)-3-hydroxy butyrate of 24 mM without requiring ATP and simultaneously recycling CoASH and NADH. This work illustrates the potential of in vitro biocatalysis to transform simple molecules into multi-functional ones.
Fil: Orrego, Alejandro H.. No especifíca;
Fil: Grazia Rubanu, Maria. No especifíca;
Fil: López, Idania L.. No especifíca;
Fil: Andrés Sanz, Daniel. No especifíca;
Fil: García-Marquina, Guillermo. No especifíca;
Fil: Pieslinger, German Eduardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina
Fil: Salassa, Luca. Universidad del País Vasco; España
Fil: López Gallego, Fernando. Universidad del País Vasco; España
description In vitro biosynthetic pathways that condense and reduce molecules through coenzyme A (CoASH) activation demand energy and redox power in the form of ATP and NAD(P)H, respectively. These coenzymes must be orthogonally recycled by ancillary reactions that consume chemicals, electricity, or light, impacting the atom economy and/or the energy consumption of the biosystem. In this work, we have exploited vinyl esters as dual acyl and electron donor substrates to synthesize β-hydroxy acids through a non-decarboxylating Claisen condensation, reduction and hydrolysis stepwise cascade, including a NADH recycling step, catalyzed by a total of 4 enzymes. Herein, the chemical energy to activate the acyl group with CoASH and the redox power for the reduction are embedded into the vinyl esters. Upon optimization, this self-sustaining cascade reached a titer of (S)-3-hydroxy butyrate of 24 mM without requiring ATP and simultaneously recycling CoASH and NADH. This work illustrates the potential of in vitro biocatalysis to transform simple molecules into multi-functional ones.
publishDate 2023
dc.date.none.fl_str_mv 2023-03
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/227763
Orrego, Alejandro H.; Grazia Rubanu, Maria; López, Idania L.; Andrés Sanz, Daniel; García-Marquina, Guillermo; et al.; ATP-Independent and Cell-Free Biosynthesis of β-Hydroxy Acids Using Vinyl Esters as Smart Substrates; Wiley VCH Verlag; Angewandte Chemie; 62; 13; 3-2023; 1-9
1433-7851
CONICET Digital
CONICET
url http://hdl.handle.net/11336/227763
identifier_str_mv Orrego, Alejandro H.; Grazia Rubanu, Maria; López, Idania L.; Andrés Sanz, Daniel; García-Marquina, Guillermo; et al.; ATP-Independent and Cell-Free Biosynthesis of β-Hydroxy Acids Using Vinyl Esters as Smart Substrates; Wiley VCH Verlag; Angewandte Chemie; 62; 13; 3-2023; 1-9
1433-7851
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1002/anie.202218312
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Wiley VCH Verlag
publisher.none.fl_str_mv Wiley VCH Verlag
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
_version_ 1846082873244778496
score 13.221938

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