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
- Institución
- Consejo Nacional de Investigaciones Científicas y Técnicas
- OAI Identificador
- oai:ri.conicet.gov.ar:11336/112531
Ver los metadatos del registro completo
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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 Técnicas2025-09-29T09:43:46Zoai: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:34982025-09-29 09:43:46.474CONICET 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) |
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CONICET Digital (CONICET) |
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Consejo Nacional de Investigaciones Científicas y Técnicas |
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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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13.070432 |