Expression and characterization of recombinant cytosolic NAD+-dependent malate dehydrogenase from Mesembryanthemum crystallinum

Autores
Tripodi, Karina Eva Josefina; Gomez Casati, Maria Eugenia; Uttaro, Antonio Domingo; Podesta, Florencio Esteban
Año de publicación
2003
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
An NAD+-dependent cytosolic malate dehydrogenase (MDH, EC 1.1.1.37) from leaves of Mesembryanthemum crystallinum in the Crassulacean Acid Metabolism (CAM) mode was cloned, expressed in E. coli and characterized. The recombinant enzyme had a subunit molecular mass of 39.5 kDa and was recognized by antibodies raised against the cytosolic MDH from Ananas comosus. Its activity showed a maximum in the pH range of 7.5-9.5. The purified MDH is highly but not completely specific for oxaloacetate, as indicated by a low activity using various other α-ketoacids as substrates. The sequence data, subunit mass and immunoreactivity suggest that the MDH that has been cloned and characterized corresponds to the cytosolic isoform. Yet, the biochemistry of this enzyme comparative with the only other CAM plant cytosolic MDH characterized so far (that of pineapple) hints at a distinct isoform being expressed in M. crystallinum leaves.
Fil: Tripodi, Karina Eva Josefina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Centro de Estudios Fotosintéticos y Bioquímicos. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Centro de Estudios Fotosintéticos y Bioquímicos; Argentina
Fil: Gomez Casati, Maria Eugenia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Centro de Estudios Fotosintéticos y Bioquímicos. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Centro de Estudios Fotosintéticos y Bioquímicos; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
Fil: Uttaro, Antonio Domingo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; Argentina
Fil: Podesta, Florencio Esteban. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Centro de Estudios Fotosintéticos y Bioquímicos. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Centro de Estudios Fotosintéticos y Bioquímicos; Argentina
Materia
Crassulacean Acid Metabolism
Malate Dehydrogenase
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:11336/79768
Acceder
id CONICETDig_07758238e94b50c5dc39f30603635f13
oai_identifier_str oai:ri.conicet.gov.ar:11336/79768
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Expression and characterization of recombinant cytosolic NAD+-dependent malate dehydrogenase from Mesembryanthemum crystallinumTripodi, Karina Eva JosefinaGomez Casati, Maria EugeniaUttaro, Antonio DomingoPodesta, Florencio EstebanCrassulacean Acid MetabolismMalate Dehydrogenasehttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1An NAD+-dependent cytosolic malate dehydrogenase (MDH, EC 1.1.1.37) from leaves of Mesembryanthemum crystallinum in the Crassulacean Acid Metabolism (CAM) mode was cloned, expressed in E. coli and characterized. The recombinant enzyme had a subunit molecular mass of 39.5 kDa and was recognized by antibodies raised against the cytosolic MDH from Ananas comosus. Its activity showed a maximum in the pH range of 7.5-9.5. The purified MDH is highly but not completely specific for oxaloacetate, as indicated by a low activity using various other α-ketoacids as substrates. The sequence data, subunit mass and immunoreactivity suggest that the MDH that has been cloned and characterized corresponds to the cytosolic isoform. Yet, the biochemistry of this enzyme comparative with the only other CAM plant cytosolic MDH characterized so far (that of pineapple) hints at a distinct isoform being expressed in M. crystallinum leaves.Fil: Tripodi, Karina Eva Josefina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Centro de Estudios Fotosintéticos y Bioquímicos. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Centro de Estudios Fotosintéticos y Bioquímicos; ArgentinaFil: Gomez Casati, Maria Eugenia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Centro de Estudios Fotosintéticos y Bioquímicos. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Centro de Estudios Fotosintéticos y Bioquímicos; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Uttaro, Antonio Domingo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; ArgentinaFil: Podesta, Florencio Esteban. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Centro de Estudios Fotosintéticos y Bioquímicos. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Centro de Estudios Fotosintéticos y Bioquímicos; ArgentinaWiley Blackwell Publishing, Inc2003-02info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/79768Tripodi, Karina Eva Josefina; Gomez Casati, Maria Eugenia; Uttaro, Antonio Domingo; Podesta, Florencio Esteban; Expression and characterization of recombinant cytosolic NAD+-dependent malate dehydrogenase from Mesembryanthemum crystallinum; Wiley Blackwell Publishing, Inc; Physiologia Plantarum; 117; 2; 2-2003; 222-2280031-9317CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1034/j.1399-3054.2003.00010.xinfo:eu-repo/semantics/altIdentifier/url/https://onlinelibrary.wiley.com/doi/abs/10.1034/j.1399-3054.2003.00010.xinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-22T11:53:38Zoai:ri.conicet.gov.ar:11336/79768instacron: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 11:53:38.965CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Expression and characterization of recombinant cytosolic NAD+-dependent malate dehydrogenase from Mesembryanthemum crystallinum
title Expression and characterization of recombinant cytosolic NAD+-dependent malate dehydrogenase from Mesembryanthemum crystallinum
spellingShingle Expression and characterization of recombinant cytosolic NAD+-dependent malate dehydrogenase from Mesembryanthemum crystallinum
Tripodi, Karina Eva Josefina
Crassulacean Acid Metabolism
Malate Dehydrogenase
title_short Expression and characterization of recombinant cytosolic NAD+-dependent malate dehydrogenase from Mesembryanthemum crystallinum
title_full Expression and characterization of recombinant cytosolic NAD+-dependent malate dehydrogenase from Mesembryanthemum crystallinum
title_fullStr Expression and characterization of recombinant cytosolic NAD+-dependent malate dehydrogenase from Mesembryanthemum crystallinum
title_full_unstemmed Expression and characterization of recombinant cytosolic NAD+-dependent malate dehydrogenase from Mesembryanthemum crystallinum
title_sort Expression and characterization of recombinant cytosolic NAD+-dependent malate dehydrogenase from Mesembryanthemum crystallinum
dc.creator.none.fl_str_mv Tripodi, Karina Eva Josefina
Gomez Casati, Maria Eugenia
Uttaro, Antonio Domingo
Podesta, Florencio Esteban
author Tripodi, Karina Eva Josefina
author_facet Tripodi, Karina Eva Josefina
Gomez Casati, Maria Eugenia
Uttaro, Antonio Domingo
Podesta, Florencio Esteban
author_role author
author2 Gomez Casati, Maria Eugenia
Uttaro, Antonio Domingo
Podesta, Florencio Esteban
author2_role author
author
author
dc.subject.none.fl_str_mv Crassulacean Acid Metabolism
Malate Dehydrogenase
topic Crassulacean Acid Metabolism
Malate Dehydrogenase
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv An NAD+-dependent cytosolic malate dehydrogenase (MDH, EC 1.1.1.37) from leaves of Mesembryanthemum crystallinum in the Crassulacean Acid Metabolism (CAM) mode was cloned, expressed in E. coli and characterized. The recombinant enzyme had a subunit molecular mass of 39.5 kDa and was recognized by antibodies raised against the cytosolic MDH from Ananas comosus. Its activity showed a maximum in the pH range of 7.5-9.5. The purified MDH is highly but not completely specific for oxaloacetate, as indicated by a low activity using various other α-ketoacids as substrates. The sequence data, subunit mass and immunoreactivity suggest that the MDH that has been cloned and characterized corresponds to the cytosolic isoform. Yet, the biochemistry of this enzyme comparative with the only other CAM plant cytosolic MDH characterized so far (that of pineapple) hints at a distinct isoform being expressed in M. crystallinum leaves.
Fil: Tripodi, Karina Eva Josefina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Centro de Estudios Fotosintéticos y Bioquímicos. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Centro de Estudios Fotosintéticos y Bioquímicos; Argentina
Fil: Gomez Casati, Maria Eugenia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Centro de Estudios Fotosintéticos y Bioquímicos. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Centro de Estudios Fotosintéticos y Bioquímicos; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
Fil: Uttaro, Antonio Domingo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; Argentina
Fil: Podesta, Florencio Esteban. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Centro de Estudios Fotosintéticos y Bioquímicos. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Centro de Estudios Fotosintéticos y Bioquímicos; Argentina
description An NAD+-dependent cytosolic malate dehydrogenase (MDH, EC 1.1.1.37) from leaves of Mesembryanthemum crystallinum in the Crassulacean Acid Metabolism (CAM) mode was cloned, expressed in E. coli and characterized. The recombinant enzyme had a subunit molecular mass of 39.5 kDa and was recognized by antibodies raised against the cytosolic MDH from Ananas comosus. Its activity showed a maximum in the pH range of 7.5-9.5. The purified MDH is highly but not completely specific for oxaloacetate, as indicated by a low activity using various other α-ketoacids as substrates. The sequence data, subunit mass and immunoreactivity suggest that the MDH that has been cloned and characterized corresponds to the cytosolic isoform. Yet, the biochemistry of this enzyme comparative with the only other CAM plant cytosolic MDH characterized so far (that of pineapple) hints at a distinct isoform being expressed in M. crystallinum leaves.
publishDate 2003
dc.date.none.fl_str_mv 2003-02
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/11336/79768
Tripodi, Karina Eva Josefina; Gomez Casati, Maria Eugenia; Uttaro, Antonio Domingo; Podesta, Florencio Esteban; Expression and characterization of recombinant cytosolic NAD+-dependent malate dehydrogenase from Mesembryanthemum crystallinum; Wiley Blackwell Publishing, Inc; Physiologia Plantarum; 117; 2; 2-2003; 222-228
0031-9317
CONICET Digital
CONICET
url http://hdl.handle.net/11336/79768
identifier_str_mv Tripodi, Karina Eva Josefina; Gomez Casati, Maria Eugenia; Uttaro, Antonio Domingo; Podesta, Florencio Esteban; Expression and characterization of recombinant cytosolic NAD+-dependent malate dehydrogenase from Mesembryanthemum crystallinum; Wiley Blackwell Publishing, Inc; Physiologia Plantarum; 117; 2; 2-2003; 222-228
0031-9317
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1034/j.1399-3054.2003.00010.x
info:eu-repo/semantics/altIdentifier/url/https://onlinelibrary.wiley.com/doi/abs/10.1034/j.1399-3054.2003.00010.x
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 Blackwell Publishing, Inc
publisher.none.fl_str_mv Wiley Blackwell Publishing, Inc
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_ 1846782232861081600
score 12.982451

Publicaciones similares