Discovery and engineering of an aldehyde tolerant 2-deoxy-d-ribose 5-phosphate aldolase (Dera) from pectobacterium atrosepticum

Autores
Haridas, Meera; Bisterfeld, Carolin; Chen, Le Min; Marsden, Stefan R.; Tonin, Fabio; Médici, Rosario; Iribarren, Adolfo Marcelo; Lewkowicz, Elizabeth Sandra; Hagedoorn, Peter-Leon; Hanefeld, Ulf; Abdelraheem, Eman
Año de publicación
2020
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
DERA (2-Deoxy-D-ribose 5-phosphate aldolase) is the only known aldolase that accepts two aldehyde substrates, which makes it an attractive catalyst for the synthesis of a chiral polyol motif that is present in several pharmaceuticals, such as atorvastatin and pravastatin. However, inactivation of the enzyme in the presence of aldehydes hinders its practical application. Whole cells of Pectobacterium atrosepticum were reported to exhibit good tolerance toward acetaldehyde and to afford 2-deoxyribose 5-phosphate with good yields. The DERA gene (PaDERA) was identified, and both the wild-type and a C49M mutant were heterologously expressed in Escherichia coli. The purification protocol was optimized and an initial biochemical characterization was conducted. Unlike other DERAs, which show a maximal activity between pH 4.0 and 7.5, PaDERA presented an optimum pH in the alkaline range between 8.0 and 9.0. This could warrant its use for specific syntheses in the future. PaDERA also displayed fourfold higher specific activity than DERA from E. coli (EcDERA) and displayed a promising acetaldehyde resistance outside the whole-cell environment. The C49M mutation, which was previously identified to increase acetaldehyde tolerance in EcDERA, also led to significant improvements in the acetaldehyde tolerance of PaDERA.
Fil: Haridas, Meera. Delft University of Technology; Países Bajos
Fil: Bisterfeld, Carolin. Delft University of Technology; Países Bajos
Fil: Chen, Le Min. Delft University of Technology; Países Bajos
Fil: Marsden, Stefan R.. Delft University of Technology; Países Bajos
Fil: Tonin, Fabio. Delft University of Technology; Países Bajos
Fil: Médici, Rosario. Delft University of Technology; Países Bajos
Fil: Iribarren, Adolfo Marcelo. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Área Química. Laboratorio de Biotransformaciones; Argentina
Fil: Lewkowicz, Elizabeth Sandra. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Área Química. Laboratorio de Biotransformaciones; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Hagedoorn, Peter-Leon. Delft University of Technology; Países Bajos
Fil: Hanefeld, Ulf. Delft University of Technology; Países Bajos
Fil: Abdelraheem, Eman. Delft University of Technology; Países Bajos
Materia
ACETALDEHYDE RESISTANCE
ALDOLASE
DERA
PECTOBACTERIUM ATROSEPTICUM
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/170004
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/170004
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Discovery and engineering of an aldehyde tolerant 2-deoxy-d-ribose 5-phosphate aldolase (Dera) from pectobacterium atrosepticumHaridas, MeeraBisterfeld, CarolinChen, Le MinMarsden, Stefan R.Tonin, FabioMédici, RosarioIribarren, Adolfo MarceloLewkowicz, Elizabeth SandraHagedoorn, Peter-LeonHanefeld, UlfAbdelraheem, EmanACETALDEHYDE RESISTANCEALDOLASEDERAPECTOBACTERIUM ATROSEPTICUMhttps://purl.org/becyt/ford/2.9https://purl.org/becyt/ford/2DERA (2-Deoxy-D-ribose 5-phosphate aldolase) is the only known aldolase that accepts two aldehyde substrates, which makes it an attractive catalyst for the synthesis of a chiral polyol motif that is present in several pharmaceuticals, such as atorvastatin and pravastatin. However, inactivation of the enzyme in the presence of aldehydes hinders its practical application. Whole cells of Pectobacterium atrosepticum were reported to exhibit good tolerance toward acetaldehyde and to afford 2-deoxyribose 5-phosphate with good yields. The DERA gene (PaDERA) was identified, and both the wild-type and a C49M mutant were heterologously expressed in Escherichia coli. The purification protocol was optimized and an initial biochemical characterization was conducted. Unlike other DERAs, which show a maximal activity between pH 4.0 and 7.5, PaDERA presented an optimum pH in the alkaline range between 8.0 and 9.0. This could warrant its use for specific syntheses in the future. PaDERA also displayed fourfold higher specific activity than DERA from E. coli (EcDERA) and displayed a promising acetaldehyde resistance outside the whole-cell environment. The C49M mutation, which was previously identified to increase acetaldehyde tolerance in EcDERA, also led to significant improvements in the acetaldehyde tolerance of PaDERA.Fil: Haridas, Meera. Delft University of Technology; Países BajosFil: Bisterfeld, Carolin. Delft University of Technology; Países BajosFil: Chen, Le Min. Delft University of Technology; Países BajosFil: Marsden, Stefan R.. Delft University of Technology; Países BajosFil: Tonin, Fabio. Delft University of Technology; Países BajosFil: Médici, Rosario. Delft University of Technology; Países BajosFil: Iribarren, Adolfo Marcelo. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Área Química. Laboratorio de Biotransformaciones; ArgentinaFil: Lewkowicz, Elizabeth Sandra. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Área Química. Laboratorio de Biotransformaciones; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Hagedoorn, Peter-Leon. Delft University of Technology; Países BajosFil: Hanefeld, Ulf. Delft University of Technology; Países BajosFil: Abdelraheem, Eman. Delft University of Technology; Países BajosMDPI2020-08info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/170004Haridas, Meera; Bisterfeld, Carolin; Chen, Le Min; Marsden, Stefan R.; Tonin, Fabio; et al.; Discovery and engineering of an aldehyde tolerant 2-deoxy-d-ribose 5-phosphate aldolase (Dera) from pectobacterium atrosepticum; MDPI; Catalysts; 10; 8; 8-2020; 1-102073-4344CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.3390/catal10080883info:eu-repo/semantics/altIdentifier/url/https://www.mdpi.com/2073-4344/10/8/883info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:01:17Zoai:ri.conicet.gov.ar:11336/170004instacron: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 10:01:17.342CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Discovery and engineering of an aldehyde tolerant 2-deoxy-d-ribose 5-phosphate aldolase (Dera) from pectobacterium atrosepticum
title Discovery and engineering of an aldehyde tolerant 2-deoxy-d-ribose 5-phosphate aldolase (Dera) from pectobacterium atrosepticum
spellingShingle Discovery and engineering of an aldehyde tolerant 2-deoxy-d-ribose 5-phosphate aldolase (Dera) from pectobacterium atrosepticum
Haridas, Meera
ACETALDEHYDE RESISTANCE
ALDOLASE
DERA
PECTOBACTERIUM ATROSEPTICUM
title_short Discovery and engineering of an aldehyde tolerant 2-deoxy-d-ribose 5-phosphate aldolase (Dera) from pectobacterium atrosepticum
title_full Discovery and engineering of an aldehyde tolerant 2-deoxy-d-ribose 5-phosphate aldolase (Dera) from pectobacterium atrosepticum
title_fullStr Discovery and engineering of an aldehyde tolerant 2-deoxy-d-ribose 5-phosphate aldolase (Dera) from pectobacterium atrosepticum
title_full_unstemmed Discovery and engineering of an aldehyde tolerant 2-deoxy-d-ribose 5-phosphate aldolase (Dera) from pectobacterium atrosepticum
title_sort Discovery and engineering of an aldehyde tolerant 2-deoxy-d-ribose 5-phosphate aldolase (Dera) from pectobacterium atrosepticum
dc.creator.none.fl_str_mv Haridas, Meera
Bisterfeld, Carolin
Chen, Le Min
Marsden, Stefan R.
Tonin, Fabio
Médici, Rosario
Iribarren, Adolfo Marcelo
Lewkowicz, Elizabeth Sandra
Hagedoorn, Peter-Leon
Hanefeld, Ulf
Abdelraheem, Eman
author Haridas, Meera
author_facet Haridas, Meera
Bisterfeld, Carolin
Chen, Le Min
Marsden, Stefan R.
Tonin, Fabio
Médici, Rosario
Iribarren, Adolfo Marcelo
Lewkowicz, Elizabeth Sandra
Hagedoorn, Peter-Leon
Hanefeld, Ulf
Abdelraheem, Eman
author_role author
author2 Bisterfeld, Carolin
Chen, Le Min
Marsden, Stefan R.
Tonin, Fabio
Médici, Rosario
Iribarren, Adolfo Marcelo
Lewkowicz, Elizabeth Sandra
Hagedoorn, Peter-Leon
Hanefeld, Ulf
Abdelraheem, Eman
author2_role author
author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv ACETALDEHYDE RESISTANCE
ALDOLASE
DERA
PECTOBACTERIUM ATROSEPTICUM
topic ACETALDEHYDE RESISTANCE
ALDOLASE
DERA
PECTOBACTERIUM ATROSEPTICUM
purl_subject.fl_str_mv https://purl.org/becyt/ford/2.9
https://purl.org/becyt/ford/2
dc.description.none.fl_txt_mv DERA (2-Deoxy-D-ribose 5-phosphate aldolase) is the only known aldolase that accepts two aldehyde substrates, which makes it an attractive catalyst for the synthesis of a chiral polyol motif that is present in several pharmaceuticals, such as atorvastatin and pravastatin. However, inactivation of the enzyme in the presence of aldehydes hinders its practical application. Whole cells of Pectobacterium atrosepticum were reported to exhibit good tolerance toward acetaldehyde and to afford 2-deoxyribose 5-phosphate with good yields. The DERA gene (PaDERA) was identified, and both the wild-type and a C49M mutant were heterologously expressed in Escherichia coli. The purification protocol was optimized and an initial biochemical characterization was conducted. Unlike other DERAs, which show a maximal activity between pH 4.0 and 7.5, PaDERA presented an optimum pH in the alkaline range between 8.0 and 9.0. This could warrant its use for specific syntheses in the future. PaDERA also displayed fourfold higher specific activity than DERA from E. coli (EcDERA) and displayed a promising acetaldehyde resistance outside the whole-cell environment. The C49M mutation, which was previously identified to increase acetaldehyde tolerance in EcDERA, also led to significant improvements in the acetaldehyde tolerance of PaDERA.
Fil: Haridas, Meera. Delft University of Technology; Países Bajos
Fil: Bisterfeld, Carolin. Delft University of Technology; Países Bajos
Fil: Chen, Le Min. Delft University of Technology; Países Bajos
Fil: Marsden, Stefan R.. Delft University of Technology; Países Bajos
Fil: Tonin, Fabio. Delft University of Technology; Países Bajos
Fil: Médici, Rosario. Delft University of Technology; Países Bajos
Fil: Iribarren, Adolfo Marcelo. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Área Química. Laboratorio de Biotransformaciones; Argentina
Fil: Lewkowicz, Elizabeth Sandra. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Área Química. Laboratorio de Biotransformaciones; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Hagedoorn, Peter-Leon. Delft University of Technology; Países Bajos
Fil: Hanefeld, Ulf. Delft University of Technology; Países Bajos
Fil: Abdelraheem, Eman. Delft University of Technology; Países Bajos
description DERA (2-Deoxy-D-ribose 5-phosphate aldolase) is the only known aldolase that accepts two aldehyde substrates, which makes it an attractive catalyst for the synthesis of a chiral polyol motif that is present in several pharmaceuticals, such as atorvastatin and pravastatin. However, inactivation of the enzyme in the presence of aldehydes hinders its practical application. Whole cells of Pectobacterium atrosepticum were reported to exhibit good tolerance toward acetaldehyde and to afford 2-deoxyribose 5-phosphate with good yields. The DERA gene (PaDERA) was identified, and both the wild-type and a C49M mutant were heterologously expressed in Escherichia coli. The purification protocol was optimized and an initial biochemical characterization was conducted. Unlike other DERAs, which show a maximal activity between pH 4.0 and 7.5, PaDERA presented an optimum pH in the alkaline range between 8.0 and 9.0. This could warrant its use for specific syntheses in the future. PaDERA also displayed fourfold higher specific activity than DERA from E. coli (EcDERA) and displayed a promising acetaldehyde resistance outside the whole-cell environment. The C49M mutation, which was previously identified to increase acetaldehyde tolerance in EcDERA, also led to significant improvements in the acetaldehyde tolerance of PaDERA.
publishDate 2020
dc.date.none.fl_str_mv 2020-08
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/11336/170004
Haridas, Meera; Bisterfeld, Carolin; Chen, Le Min; Marsden, Stefan R.; Tonin, Fabio; et al.; Discovery and engineering of an aldehyde tolerant 2-deoxy-d-ribose 5-phosphate aldolase (Dera) from pectobacterium atrosepticum; MDPI; Catalysts; 10; 8; 8-2020; 1-10
2073-4344
CONICET Digital
CONICET
url http://hdl.handle.net/11336/170004
identifier_str_mv Haridas, Meera; Bisterfeld, Carolin; Chen, Le Min; Marsden, Stefan R.; Tonin, Fabio; et al.; Discovery and engineering of an aldehyde tolerant 2-deoxy-d-ribose 5-phosphate aldolase (Dera) from pectobacterium atrosepticum; MDPI; Catalysts; 10; 8; 8-2020; 1-10
2073-4344
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.3390/catal10080883
info:eu-repo/semantics/altIdentifier/url/https://www.mdpi.com/2073-4344/10/8/883
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv MDPI
publisher.none.fl_str_mv MDPI
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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