Alteration of the Route to Menaquinone towards Isochorismate-Derived Metabolites

Autores
Fries, Alexander Erich; Mazzaferro, Laura; Grüning, Björn; Bisel, Philippe; Stibal, Karin; Buchholz, Patrick C. F.; Pleiss, Jürgen; Sprenger, Georg A.; Müller, Michael
Año de publicación
2019
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Chorismate and isochorismate constitute branch-point intermediates in the biosynthesis of many aromatic metabolites in microorganisms and plants. To obtain unnatural compounds, we modified the route to menaquinone in Escherichia coli. We propose a model for the binding of isochorismate to the active site of MenD ((1R,2S, 5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxycyclohex-3-ene-1-carboxylate (SEPHCHC) synthase) that explains the outcome of the native reaction with α-ketoglutarate. We have rationally designed variants of MenD for the conversion of several isochorismate analogues. The double-variant Asn117Arg–Leu478Thr preferentially converts (5S,6S)-5,6-dihydroxycyclohexa-1,3-diene-1-carboxylate (2,3-trans-CHD), the hydrolysis product of isochorismate, with a >70-fold higher ratio than that for the wild type. The single-variant Arg107Ile uses (5S,6S)-6-amino-5-hydroxycyclohexa-1,3-diene-1-carboxylate (2,3-trans-CHA) as substrate with >6-fold conversion compared to wild-type MenD. The novel compounds have been made accessible in vivo (up to 5.3 g L−1). Unexpectedly, as the identified residues such as Arg107 are highly conserved (>94 %), some of the designed variations can be found in wild-type SEPHCHC synthases from other bacteria (Arg107Lys, 0.3 %). This raises the question for the possible natural occurrence of as yet unexplored branches of the shikimate pathway.
Fil: Fries, Alexander Erich. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Ciencias de la Tierra y Ambientales de La Pampa. Universidad Nacional de La Pampa. Facultad de Ciencias Exactas y Naturales. Instituto de Ciencias de la Tierra y Ambientales de La Pampa; Argentina. Albert Ludwigs University of Freiburg; Alemania
Fil: Mazzaferro, Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Ciencias de la Tierra y Ambientales de La Pampa. Universidad Nacional de La Pampa. Facultad de Ciencias Exactas y Naturales. Instituto de Ciencias de la Tierra y Ambientales de La Pampa; Argentina. Albert Ludwigs University of Freiburg; Alemania
Fil: Grüning, Björn. Albert Ludwigs University of Freiburg; Alemania
Fil: Bisel, Philippe. Albert Ludwigs University of Freiburg; Alemania
Fil: Stibal, Karin. Albert Ludwigs University of Freiburg; Alemania
Fil: Buchholz, Patrick C. F.. University of Stuttgart; Alemania
Fil: Pleiss, Jürgen. Universität Stuttgart;
Fil: Sprenger, Georg A.. Universität Stuttgart;
Fil: Müller, Michael. Albert Ludwigs University of Freiburg; Alemania
Materia
DIVERSITY-ORIENTED SYNTHESIS
ENZYME CATALYSIS
METABOLIC ENGINEERING
SYNTHETIC BIOLOGY
THIAMINE DIPHOSPHATE
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:11336/112455
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/112455
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Alteration of the Route to Menaquinone towards Isochorismate-Derived MetabolitesFries, Alexander ErichMazzaferro, LauraGrüning, BjörnBisel, PhilippeStibal, KarinBuchholz, Patrick C. F.Pleiss, JürgenSprenger, Georg A.Müller, MichaelDIVERSITY-ORIENTED SYNTHESISENZYME CATALYSISMETABOLIC ENGINEERINGSYNTHETIC BIOLOGYTHIAMINE DIPHOSPHATEhttps://purl.org/becyt/ford/2.9https://purl.org/becyt/ford/2Chorismate and isochorismate constitute branch-point intermediates in the biosynthesis of many aromatic metabolites in microorganisms and plants. To obtain unnatural compounds, we modified the route to menaquinone in Escherichia coli. We propose a model for the binding of isochorismate to the active site of MenD ((1R,2S, 5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxycyclohex-3-ene-1-carboxylate (SEPHCHC) synthase) that explains the outcome of the native reaction with α-ketoglutarate. We have rationally designed variants of MenD for the conversion of several isochorismate analogues. The double-variant Asn117Arg–Leu478Thr preferentially converts (5S,6S)-5,6-dihydroxycyclohexa-1,3-diene-1-carboxylate (2,3-trans-CHD), the hydrolysis product of isochorismate, with a >70-fold higher ratio than that for the wild type. The single-variant Arg107Ile uses (5S,6S)-6-amino-5-hydroxycyclohexa-1,3-diene-1-carboxylate (2,3-trans-CHA) as substrate with >6-fold conversion compared to wild-type MenD. The novel compounds have been made accessible in vivo (up to 5.3 g L−1). Unexpectedly, as the identified residues such as Arg107 are highly conserved (>94 %), some of the designed variations can be found in wild-type SEPHCHC synthases from other bacteria (Arg107Lys, 0.3 %). This raises the question for the possible natural occurrence of as yet unexplored branches of the shikimate pathway.Fil: Fries, Alexander Erich. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Ciencias de la Tierra y Ambientales de La Pampa. Universidad Nacional de La Pampa. Facultad de Ciencias Exactas y Naturales. Instituto de Ciencias de la Tierra y Ambientales de La Pampa; Argentina. Albert Ludwigs University of Freiburg; AlemaniaFil: Mazzaferro, Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Ciencias de la Tierra y Ambientales de La Pampa. Universidad Nacional de La Pampa. Facultad de Ciencias Exactas y Naturales. Instituto de Ciencias de la Tierra y Ambientales de La Pampa; Argentina. Albert Ludwigs University of Freiburg; AlemaniaFil: Grüning, Björn. Albert Ludwigs University of Freiburg; AlemaniaFil: Bisel, Philippe. Albert Ludwigs University of Freiburg; AlemaniaFil: Stibal, Karin. Albert Ludwigs University of Freiburg; AlemaniaFil: Buchholz, Patrick C. F.. University of Stuttgart; AlemaniaFil: Pleiss, Jürgen. Universität Stuttgart;Fil: Sprenger, Georg A.. Universität Stuttgart;Fil: Müller, Michael. Albert Ludwigs University of Freiburg; AlemaniaWiley VCH Verlag2019-07info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/112455Fries, Alexander Erich; Mazzaferro, Laura; Grüning, Björn; Bisel, Philippe; Stibal, Karin; et al.; Alteration of the Route to Menaquinone towards Isochorismate-Derived Metabolites; Wiley VCH Verlag; Chembiochem; 20; 13; 7-2019; 1672-16771439-4227CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1002/cbic.201900050info:eu-repo/semantics/altIdentifier/url/https://chemistry-europe.onlinelibrary.wiley.com/doi/abs/10.1002/cbic.201900050info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-nd/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:14:25Zoai:ri.conicet.gov.ar:11336/112455instacron: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:34982025-09-29 10:14:25.873CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Alteration of the Route to Menaquinone towards Isochorismate-Derived Metabolites
title Alteration of the Route to Menaquinone towards Isochorismate-Derived Metabolites
spellingShingle Alteration of the Route to Menaquinone towards Isochorismate-Derived Metabolites
Fries, Alexander Erich
DIVERSITY-ORIENTED SYNTHESIS
ENZYME CATALYSIS
METABOLIC ENGINEERING
SYNTHETIC BIOLOGY
THIAMINE DIPHOSPHATE
title_short Alteration of the Route to Menaquinone towards Isochorismate-Derived Metabolites
title_full Alteration of the Route to Menaquinone towards Isochorismate-Derived Metabolites
title_fullStr Alteration of the Route to Menaquinone towards Isochorismate-Derived Metabolites
title_full_unstemmed Alteration of the Route to Menaquinone towards Isochorismate-Derived Metabolites
title_sort Alteration of the Route to Menaquinone towards Isochorismate-Derived Metabolites
dc.creator.none.fl_str_mv Fries, Alexander Erich
Mazzaferro, Laura
Grüning, Björn
Bisel, Philippe
Stibal, Karin
Buchholz, Patrick C. F.
Pleiss, Jürgen
Sprenger, Georg A.
Müller, Michael
author Fries, Alexander Erich
author_facet Fries, Alexander Erich
Mazzaferro, Laura
Grüning, Björn
Bisel, Philippe
Stibal, Karin
Buchholz, Patrick C. F.
Pleiss, Jürgen
Sprenger, Georg A.
Müller, Michael
author_role author
author2 Mazzaferro, Laura
Grüning, Björn
Bisel, Philippe
Stibal, Karin
Buchholz, Patrick C. F.
Pleiss, Jürgen
Sprenger, Georg A.
Müller, Michael
author2_role author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv DIVERSITY-ORIENTED SYNTHESIS
ENZYME CATALYSIS
METABOLIC ENGINEERING
SYNTHETIC BIOLOGY
THIAMINE DIPHOSPHATE
topic DIVERSITY-ORIENTED SYNTHESIS
ENZYME CATALYSIS
METABOLIC ENGINEERING
SYNTHETIC BIOLOGY
THIAMINE DIPHOSPHATE
purl_subject.fl_str_mv https://purl.org/becyt/ford/2.9
https://purl.org/becyt/ford/2
dc.description.none.fl_txt_mv Chorismate and isochorismate constitute branch-point intermediates in the biosynthesis of many aromatic metabolites in microorganisms and plants. To obtain unnatural compounds, we modified the route to menaquinone in Escherichia coli. We propose a model for the binding of isochorismate to the active site of MenD ((1R,2S, 5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxycyclohex-3-ene-1-carboxylate (SEPHCHC) synthase) that explains the outcome of the native reaction with α-ketoglutarate. We have rationally designed variants of MenD for the conversion of several isochorismate analogues. The double-variant Asn117Arg–Leu478Thr preferentially converts (5S,6S)-5,6-dihydroxycyclohexa-1,3-diene-1-carboxylate (2,3-trans-CHD), the hydrolysis product of isochorismate, with a >70-fold higher ratio than that for the wild type. The single-variant Arg107Ile uses (5S,6S)-6-amino-5-hydroxycyclohexa-1,3-diene-1-carboxylate (2,3-trans-CHA) as substrate with >6-fold conversion compared to wild-type MenD. The novel compounds have been made accessible in vivo (up to 5.3 g L−1). Unexpectedly, as the identified residues such as Arg107 are highly conserved (>94 %), some of the designed variations can be found in wild-type SEPHCHC synthases from other bacteria (Arg107Lys, 0.3 %). This raises the question for the possible natural occurrence of as yet unexplored branches of the shikimate pathway.
Fil: Fries, Alexander Erich. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Ciencias de la Tierra y Ambientales de La Pampa. Universidad Nacional de La Pampa. Facultad de Ciencias Exactas y Naturales. Instituto de Ciencias de la Tierra y Ambientales de La Pampa; Argentina. Albert Ludwigs University of Freiburg; Alemania
Fil: Mazzaferro, Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Ciencias de la Tierra y Ambientales de La Pampa. Universidad Nacional de La Pampa. Facultad de Ciencias Exactas y Naturales. Instituto de Ciencias de la Tierra y Ambientales de La Pampa; Argentina. Albert Ludwigs University of Freiburg; Alemania
Fil: Grüning, Björn. Albert Ludwigs University of Freiburg; Alemania
Fil: Bisel, Philippe. Albert Ludwigs University of Freiburg; Alemania
Fil: Stibal, Karin. Albert Ludwigs University of Freiburg; Alemania
Fil: Buchholz, Patrick C. F.. University of Stuttgart; Alemania
Fil: Pleiss, Jürgen. Universität Stuttgart;
Fil: Sprenger, Georg A.. Universität Stuttgart;
Fil: Müller, Michael. Albert Ludwigs University of Freiburg; Alemania
description Chorismate and isochorismate constitute branch-point intermediates in the biosynthesis of many aromatic metabolites in microorganisms and plants. To obtain unnatural compounds, we modified the route to menaquinone in Escherichia coli. We propose a model for the binding of isochorismate to the active site of MenD ((1R,2S, 5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxycyclohex-3-ene-1-carboxylate (SEPHCHC) synthase) that explains the outcome of the native reaction with α-ketoglutarate. We have rationally designed variants of MenD for the conversion of several isochorismate analogues. The double-variant Asn117Arg–Leu478Thr preferentially converts (5S,6S)-5,6-dihydroxycyclohexa-1,3-diene-1-carboxylate (2,3-trans-CHD), the hydrolysis product of isochorismate, with a >70-fold higher ratio than that for the wild type. The single-variant Arg107Ile uses (5S,6S)-6-amino-5-hydroxycyclohexa-1,3-diene-1-carboxylate (2,3-trans-CHA) as substrate with >6-fold conversion compared to wild-type MenD. The novel compounds have been made accessible in vivo (up to 5.3 g L−1). Unexpectedly, as the identified residues such as Arg107 are highly conserved (>94 %), some of the designed variations can be found in wild-type SEPHCHC synthases from other bacteria (Arg107Lys, 0.3 %). This raises the question for the possible natural occurrence of as yet unexplored branches of the shikimate pathway.
publishDate 2019
dc.date.none.fl_str_mv 2019-07
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/11336/112455
Fries, Alexander Erich; Mazzaferro, Laura; Grüning, Björn; Bisel, Philippe; Stibal, Karin; et al.; Alteration of the Route to Menaquinone towards Isochorismate-Derived Metabolites; Wiley VCH Verlag; Chembiochem; 20; 13; 7-2019; 1672-1677
1439-4227
CONICET Digital
CONICET
url http://hdl.handle.net/11336/112455
identifier_str_mv Fries, Alexander Erich; Mazzaferro, Laura; Grüning, Björn; Bisel, Philippe; Stibal, Karin; et al.; Alteration of the Route to Menaquinone towards Isochorismate-Derived Metabolites; Wiley VCH Verlag; Chembiochem; 20; 13; 7-2019; 1672-1677
1439-4227
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/cbic.201900050
info:eu-repo/semantics/altIdentifier/url/https://chemistry-europe.onlinelibrary.wiley.com/doi/abs/10.1002/cbic.201900050
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
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score 13.070432

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