Structural and Mutagenesis Studies of the Thiamine-Dependent, Ketone-Accepting YerE from Pseudomonas protegens

Autores
Hampel, Sabrina; Steitz, Jan Patrick; Baierl, Anna; Lehwald, Patrizia; Wiesli, Luzia; Richter, Michael; Fries, Alexander Erich; Pohl, Martina; Schneider, Gunter; Dobritzsch, Doreen; Müller, Michael
Año de publicación
2018
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
A wide range of thiamine diphosphate (ThDP)‐dependent enzymes catalyze the benzoin‐type carboligation of pyruvate with aldehydes. A few ThDP‐dependent enzymes, such as YerE from Yersinia pseudotuberculosis (YpYerE), are known to accept ketones as acceptor substrates. Catalysis by YpYerE gives access to chiral tertiary alcohols, a group of products difficult to obtain in an enantioenriched form by other means. Hence, knowledge of the three‐dimensional structure of the enzyme is crucial to identify structure–activity relationships. However, YpYerE has yet to be crystallized, despite several attempts. Herein, we show that a homologue of YpYerE, namely, PpYerE from Pseudomonas protegens (59 % amino acid identity), displays similar catalytic activity: benzaldehyde and its derivatives as well as ketones are converted into chiral 2‐hydroxy ketones by using pyruvate as a donor. To enable comparison of aldehyde‐ and ketone‐accepting enzymes and to guide site‐directed mutagenesis studies, PpYerE was crystallized and its structure was determined to a resolution of 1.55 Å.
Fil: Hampel, Sabrina. Albert Ludwigs University of Freiburg; Alemania
Fil: Steitz, Jan Patrick. Albert Ludwigs University of Freiburg; Alemania
Fil: Baierl, Anna. IBG-1: Biotechnology. Forschungszentrum Jülich GmbH Wilhelm-Johnen Straße; Alemania
Fil: Lehwald, Patrizia. Albert Ludwigs University of Freiburg; Alemania
Fil: Wiesli, Luzia. Swiss Federal Laboratories For Materials Science And Technology; Suiza
Fil: Richter, Michael. Swiss Federal Laboratories For Materials Science And Technology; Suiza. Fraunhofer Institute for Interfacial Engineering and Biotechnology IGB, Branch BioCa; Alemania
Fil: Fries, Alexander Erich. Albert Ludwigs University of Freiburg; Alemania. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Pohl, Martina. IBG-1: Biotechnology. Forschungszentrum Jülich GmbH Wilhelm-Johnen Straße; Alemania
Fil: Schneider, Gunter. Karolinska Huddinge Hospital. Karolinska Institutet; Suecia
Fil: Dobritzsch, Doreen. Uppsala Universitet; Suecia
Fil: Müller, Michael. Albert Ludwigs University of Freiburg; Alemania
Materia
ASYMMETRIC SYNTHESIS
BIOCATALYSIS
BIOSYNTHESIS
C−C COUPLING
TERTIARY ALCOHOLS
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/112531
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/112531
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Structural and Mutagenesis Studies of the Thiamine-Dependent, Ketone-Accepting YerE from Pseudomonas protegensHampel, SabrinaSteitz, Jan PatrickBaierl, AnnaLehwald, PatriziaWiesli, LuziaRichter, MichaelFries, Alexander ErichPohl, MartinaSchneider, GunterDobritzsch, DoreenMüller, MichaelASYMMETRIC SYNTHESISBIOCATALYSISBIOSYNTHESISC−C COUPLINGTERTIARY ALCOHOLShttps://purl.org/becyt/ford/2.9https://purl.org/becyt/ford/2A wide range of thiamine diphosphate (ThDP)‐dependent enzymes catalyze the benzoin‐type carboligation of pyruvate with aldehydes. A few ThDP‐dependent enzymes, such as YerE from Yersinia pseudotuberculosis (YpYerE), are known to accept ketones as acceptor substrates. Catalysis by YpYerE gives access to chiral tertiary alcohols, a group of products difficult to obtain in an enantioenriched form by other means. Hence, knowledge of the three‐dimensional structure of the enzyme is crucial to identify structure–activity relationships. However, YpYerE has yet to be crystallized, despite several attempts. Herein, we show that a homologue of YpYerE, namely, PpYerE from Pseudomonas protegens (59 % amino acid identity), displays similar catalytic activity: benzaldehyde and its derivatives as well as ketones are converted into chiral 2‐hydroxy ketones by using pyruvate as a donor. To enable comparison of aldehyde‐ and ketone‐accepting enzymes and to guide site‐directed mutagenesis studies, PpYerE was crystallized and its structure was determined to a resolution of 1.55 Å.Fil: Hampel, Sabrina. Albert Ludwigs University of Freiburg; AlemaniaFil: Steitz, Jan Patrick. Albert Ludwigs University of Freiburg; AlemaniaFil: Baierl, Anna. IBG-1: Biotechnology. Forschungszentrum Jülich GmbH Wilhelm-Johnen Straße; AlemaniaFil: Lehwald, Patrizia. Albert Ludwigs University of Freiburg; AlemaniaFil: Wiesli, Luzia. Swiss Federal Laboratories For Materials Science And Technology; SuizaFil: Richter, Michael. Swiss Federal Laboratories For Materials Science And Technology; Suiza. Fraunhofer Institute for Interfacial Engineering and Biotechnology IGB, Branch BioCa; AlemaniaFil: Fries, Alexander Erich. Albert Ludwigs University of Freiburg; Alemania. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Pohl, Martina. IBG-1: Biotechnology. Forschungszentrum Jülich GmbH Wilhelm-Johnen Straße; AlemaniaFil: Schneider, Gunter. Karolinska Huddinge Hospital. Karolinska Institutet; SueciaFil: Dobritzsch, Doreen. Uppsala Universitet; SueciaFil: Müller, Michael. Albert Ludwigs University of Freiburg; AlemaniaWiley VCH Verlag2018-11info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/112531Hampel, Sabrina; Steitz, Jan Patrick; Baierl, Anna; Lehwald, Patrizia; Wiesli, Luzia; et al.; Structural and Mutagenesis Studies of the Thiamine-Dependent, Ketone-Accepting YerE from Pseudomonas protegens; Wiley VCH Verlag; Chembiochem; 19; 21; 11-2018; 2283-22921439-4227CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1002/cbic.201800325info:eu-repo/semantics/altIdentifier/url/https://chemistry-europe.onlinelibrary.wiley.com/doi/abs/10.1002/cbic.201800325info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-22T11:12:28Zoai:ri.conicet.gov.ar:11336/112531instacron: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:34982025-10-22 11:12:28.325CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Structural and Mutagenesis Studies of the Thiamine-Dependent, Ketone-Accepting YerE from Pseudomonas protegens
title Structural and Mutagenesis Studies of the Thiamine-Dependent, Ketone-Accepting YerE from Pseudomonas protegens
spellingShingle Structural and Mutagenesis Studies of the Thiamine-Dependent, Ketone-Accepting YerE from Pseudomonas protegens
Hampel, Sabrina
ASYMMETRIC SYNTHESIS
BIOCATALYSIS
BIOSYNTHESIS
C−C COUPLING
TERTIARY ALCOHOLS
title_short Structural and Mutagenesis Studies of the Thiamine-Dependent, Ketone-Accepting YerE from Pseudomonas protegens
title_full Structural and Mutagenesis Studies of the Thiamine-Dependent, Ketone-Accepting YerE from Pseudomonas protegens
title_fullStr Structural and Mutagenesis Studies of the Thiamine-Dependent, Ketone-Accepting YerE from Pseudomonas protegens
title_full_unstemmed Structural and Mutagenesis Studies of the Thiamine-Dependent, Ketone-Accepting YerE from Pseudomonas protegens
title_sort Structural and Mutagenesis Studies of the Thiamine-Dependent, Ketone-Accepting YerE from Pseudomonas protegens
dc.creator.none.fl_str_mv Hampel, Sabrina
Steitz, Jan Patrick
Baierl, Anna
Lehwald, Patrizia
Wiesli, Luzia
Richter, Michael
Fries, Alexander Erich
Pohl, Martina
Schneider, Gunter
Dobritzsch, Doreen
Müller, Michael
author Hampel, Sabrina
author_facet Hampel, Sabrina
Steitz, Jan Patrick
Baierl, Anna
Lehwald, Patrizia
Wiesli, Luzia
Richter, Michael
Fries, Alexander Erich
Pohl, Martina
Schneider, Gunter
Dobritzsch, Doreen
Müller, Michael
author_role author
author2 Steitz, Jan Patrick
Baierl, Anna
Lehwald, Patrizia
Wiesli, Luzia
Richter, Michael
Fries, Alexander Erich
Pohl, Martina
Schneider, Gunter
Dobritzsch, Doreen
Müller, Michael
author2_role author
author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv ASYMMETRIC SYNTHESIS
BIOCATALYSIS
BIOSYNTHESIS
C−C COUPLING
TERTIARY ALCOHOLS
topic ASYMMETRIC SYNTHESIS
BIOCATALYSIS
BIOSYNTHESIS
C−C COUPLING
TERTIARY ALCOHOLS
purl_subject.fl_str_mv https://purl.org/becyt/ford/2.9
https://purl.org/becyt/ford/2
dc.description.none.fl_txt_mv A wide range of thiamine diphosphate (ThDP)‐dependent enzymes catalyze the benzoin‐type carboligation of pyruvate with aldehydes. A few ThDP‐dependent enzymes, such as YerE from Yersinia pseudotuberculosis (YpYerE), are known to accept ketones as acceptor substrates. Catalysis by YpYerE gives access to chiral tertiary alcohols, a group of products difficult to obtain in an enantioenriched form by other means. Hence, knowledge of the three‐dimensional structure of the enzyme is crucial to identify structure–activity relationships. However, YpYerE has yet to be crystallized, despite several attempts. Herein, we show that a homologue of YpYerE, namely, PpYerE from Pseudomonas protegens (59 % amino acid identity), displays similar catalytic activity: benzaldehyde and its derivatives as well as ketones are converted into chiral 2‐hydroxy ketones by using pyruvate as a donor. To enable comparison of aldehyde‐ and ketone‐accepting enzymes and to guide site‐directed mutagenesis studies, PpYerE was crystallized and its structure was determined to a resolution of 1.55 Å.
Fil: Hampel, Sabrina. Albert Ludwigs University of Freiburg; Alemania
Fil: Steitz, Jan Patrick. Albert Ludwigs University of Freiburg; Alemania
Fil: Baierl, Anna. IBG-1: Biotechnology. Forschungszentrum Jülich GmbH Wilhelm-Johnen Straße; Alemania
Fil: Lehwald, Patrizia. Albert Ludwigs University of Freiburg; Alemania
Fil: Wiesli, Luzia. Swiss Federal Laboratories For Materials Science And Technology; Suiza
Fil: Richter, Michael. Swiss Federal Laboratories For Materials Science And Technology; Suiza. Fraunhofer Institute for Interfacial Engineering and Biotechnology IGB, Branch BioCa; Alemania
Fil: Fries, Alexander Erich. Albert Ludwigs University of Freiburg; Alemania. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Pohl, Martina. IBG-1: Biotechnology. Forschungszentrum Jülich GmbH Wilhelm-Johnen Straße; Alemania
Fil: Schneider, Gunter. Karolinska Huddinge Hospital. Karolinska Institutet; Suecia
Fil: Dobritzsch, Doreen. Uppsala Universitet; Suecia
Fil: Müller, Michael. Albert Ludwigs University of Freiburg; Alemania
description A wide range of thiamine diphosphate (ThDP)‐dependent enzymes catalyze the benzoin‐type carboligation of pyruvate with aldehydes. A few ThDP‐dependent enzymes, such as YerE from Yersinia pseudotuberculosis (YpYerE), are known to accept ketones as acceptor substrates. Catalysis by YpYerE gives access to chiral tertiary alcohols, a group of products difficult to obtain in an enantioenriched form by other means. Hence, knowledge of the three‐dimensional structure of the enzyme is crucial to identify structure–activity relationships. However, YpYerE has yet to be crystallized, despite several attempts. Herein, we show that a homologue of YpYerE, namely, PpYerE from Pseudomonas protegens (59 % amino acid identity), displays similar catalytic activity: benzaldehyde and its derivatives as well as ketones are converted into chiral 2‐hydroxy ketones by using pyruvate as a donor. To enable comparison of aldehyde‐ and ketone‐accepting enzymes and to guide site‐directed mutagenesis studies, PpYerE was crystallized and its structure was determined to a resolution of 1.55 Å.
publishDate 2018
dc.date.none.fl_str_mv 2018-11
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/11336/112531
Hampel, Sabrina; Steitz, Jan Patrick; Baierl, Anna; Lehwald, Patrizia; Wiesli, Luzia; et al.; Structural and Mutagenesis Studies of the Thiamine-Dependent, Ketone-Accepting YerE from Pseudomonas protegens; Wiley VCH Verlag; Chembiochem; 19; 21; 11-2018; 2283-2292
1439-4227
CONICET Digital
CONICET
url http://hdl.handle.net/11336/112531
identifier_str_mv Hampel, Sabrina; Steitz, Jan Patrick; Baierl, Anna; Lehwald, Patrizia; Wiesli, Luzia; et al.; Structural and Mutagenesis Studies of the Thiamine-Dependent, Ketone-Accepting YerE from Pseudomonas protegens; Wiley VCH Verlag; Chembiochem; 19; 21; 11-2018; 2283-2292
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.201800325
info:eu-repo/semantics/altIdentifier/url/https://chemistry-europe.onlinelibrary.wiley.com/doi/abs/10.1002/cbic.201800325
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/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 12.982451

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