Heterologous expression and kinetic characterization of the α β and αβ blend of the PPi-dependent phosphofructokinase from Citrus sinensis
- Autores
- Muchut, Robertino José; Piattoni, Claudia Vanesa; Margarit, Ezequiel; Tripodi, Karina Eva Josefina; Podesta, Florencio Esteban; Iglesias, Alberto Alvaro
- Año de publicación
- 2019
- Idioma
- inglés
- Tipo de recurso
- artículo
- Estado
- versión publicada
- Descripción
- This work reports the molecular cloning and heterologous expression of the genes coding for α and β subunits of pyrophosphate-dependent phosphofructokinase (PPi-PFK) from orange. When expressed individually, both recombinant subunits were produced as highly purified monomeric proteins able to phosphorylate fructose-6-phosphate at the expenses of PPi (specific activity of 0.075 and 0.017 units. mg −1 for α and β subunits, respectively). On the other hand, co-expression rendered a α 3 β 3 hexamer with specific activity three orders of magnitude higher than the single subunits. All the conformations of the enzyme were characterized with respect to its kinetic properties and sensitivity to the regulator fructose-2,6-bisphosphate. A thorough review of current knowledge on the matter indicates that this is the first report of the recombinant production of active plant PPi-PFK and the characterization of its different conformations. This is a main contribution for future studies focused to better understand the enzyme properties and how it accomplishes its relevant role in plant metabolism.
Fil: Muchut, Robertino José. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; Argentina
Fil: Piattoni, Claudia Vanesa. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; Argentina
Fil: Margarit, Ezequiel. 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: 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: 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
Fil: Iglesias, Alberto Alvaro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; Argentina - Materia
-
CITRUS SINENSIS
FRUCTOSE-2,6-BISPHOSPHATE
HETERO-HEXAMER
ORANGE
PPI-DEPENDENT PHOSPHOFRUCTOKINASE
RECOMBINANT PROTEIN - Nivel de accesibilidad
- acceso abierto
- Condiciones de uso
- https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
- Repositorio
- Institución
- Consejo Nacional de Investigaciones Científicas y Técnicas
- OAI Identificador
- oai:ri.conicet.gov.ar:11336/148468
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CONICET Digital (CONICET) |
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Heterologous expression and kinetic characterization of the α β and αβ blend of the PPi-dependent phosphofructokinase from Citrus sinensisMuchut, Robertino JoséPiattoni, Claudia VanesaMargarit, EzequielTripodi, Karina Eva JosefinaPodesta, Florencio EstebanIglesias, Alberto AlvaroCITRUS SINENSISFRUCTOSE-2,6-BISPHOSPHATEHETERO-HEXAMERORANGEPPI-DEPENDENT PHOSPHOFRUCTOKINASERECOMBINANT PROTEINhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1This work reports the molecular cloning and heterologous expression of the genes coding for α and β subunits of pyrophosphate-dependent phosphofructokinase (PPi-PFK) from orange. When expressed individually, both recombinant subunits were produced as highly purified monomeric proteins able to phosphorylate fructose-6-phosphate at the expenses of PPi (specific activity of 0.075 and 0.017 units. mg −1 for α and β subunits, respectively). On the other hand, co-expression rendered a α 3 β 3 hexamer with specific activity three orders of magnitude higher than the single subunits. All the conformations of the enzyme were characterized with respect to its kinetic properties and sensitivity to the regulator fructose-2,6-bisphosphate. A thorough review of current knowledge on the matter indicates that this is the first report of the recombinant production of active plant PPi-PFK and the characterization of its different conformations. This is a main contribution for future studies focused to better understand the enzyme properties and how it accomplishes its relevant role in plant metabolism.Fil: Muchut, Robertino José. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; ArgentinaFil: Piattoni, Claudia Vanesa. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; ArgentinaFil: Margarit, Ezequiel. 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: 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: 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; ArgentinaFil: Iglesias, Alberto Alvaro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; ArgentinaElsevier Ireland2019-03info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/148468Muchut, Robertino José; Piattoni, Claudia Vanesa; Margarit, Ezequiel; Tripodi, Karina Eva Josefina; Podesta, Florencio Esteban; et al.; Heterologous expression and kinetic characterization of the α β and αβ blend of the PPi-dependent phosphofructokinase from Citrus sinensis; Elsevier Ireland; Plant Science; 280; 3-2019; 348-3540168-9452CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/abs/pii/S0168945218311075info:eu-repo/semantics/altIdentifier/doi/10.1016/j.plantsci.2018.12.012info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-nd/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T09:44:41Zoai:ri.conicet.gov.ar:11336/148468instacron: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 09:44:41.355CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse |
dc.title.none.fl_str_mv |
Heterologous expression and kinetic characterization of the α β and αβ blend of the PPi-dependent phosphofructokinase from Citrus sinensis |
title |
Heterologous expression and kinetic characterization of the α β and αβ blend of the PPi-dependent phosphofructokinase from Citrus sinensis |
spellingShingle |
Heterologous expression and kinetic characterization of the α β and αβ blend of the PPi-dependent phosphofructokinase from Citrus sinensis Muchut, Robertino José CITRUS SINENSIS FRUCTOSE-2,6-BISPHOSPHATE HETERO-HEXAMER ORANGE PPI-DEPENDENT PHOSPHOFRUCTOKINASE RECOMBINANT PROTEIN |
title_short |
Heterologous expression and kinetic characterization of the α β and αβ blend of the PPi-dependent phosphofructokinase from Citrus sinensis |
title_full |
Heterologous expression and kinetic characterization of the α β and αβ blend of the PPi-dependent phosphofructokinase from Citrus sinensis |
title_fullStr |
Heterologous expression and kinetic characterization of the α β and αβ blend of the PPi-dependent phosphofructokinase from Citrus sinensis |
title_full_unstemmed |
Heterologous expression and kinetic characterization of the α β and αβ blend of the PPi-dependent phosphofructokinase from Citrus sinensis |
title_sort |
Heterologous expression and kinetic characterization of the α β and αβ blend of the PPi-dependent phosphofructokinase from Citrus sinensis |
dc.creator.none.fl_str_mv |
Muchut, Robertino José Piattoni, Claudia Vanesa Margarit, Ezequiel Tripodi, Karina Eva Josefina Podesta, Florencio Esteban Iglesias, Alberto Alvaro |
author |
Muchut, Robertino José |
author_facet |
Muchut, Robertino José Piattoni, Claudia Vanesa Margarit, Ezequiel Tripodi, Karina Eva Josefina Podesta, Florencio Esteban Iglesias, Alberto Alvaro |
author_role |
author |
author2 |
Piattoni, Claudia Vanesa Margarit, Ezequiel Tripodi, Karina Eva Josefina Podesta, Florencio Esteban Iglesias, Alberto Alvaro |
author2_role |
author author author author author |
dc.subject.none.fl_str_mv |
CITRUS SINENSIS FRUCTOSE-2,6-BISPHOSPHATE HETERO-HEXAMER ORANGE PPI-DEPENDENT PHOSPHOFRUCTOKINASE RECOMBINANT PROTEIN |
topic |
CITRUS SINENSIS FRUCTOSE-2,6-BISPHOSPHATE HETERO-HEXAMER ORANGE PPI-DEPENDENT PHOSPHOFRUCTOKINASE RECOMBINANT PROTEIN |
purl_subject.fl_str_mv |
https://purl.org/becyt/ford/1.6 https://purl.org/becyt/ford/1 |
dc.description.none.fl_txt_mv |
This work reports the molecular cloning and heterologous expression of the genes coding for α and β subunits of pyrophosphate-dependent phosphofructokinase (PPi-PFK) from orange. When expressed individually, both recombinant subunits were produced as highly purified monomeric proteins able to phosphorylate fructose-6-phosphate at the expenses of PPi (specific activity of 0.075 and 0.017 units. mg −1 for α and β subunits, respectively). On the other hand, co-expression rendered a α 3 β 3 hexamer with specific activity three orders of magnitude higher than the single subunits. All the conformations of the enzyme were characterized with respect to its kinetic properties and sensitivity to the regulator fructose-2,6-bisphosphate. A thorough review of current knowledge on the matter indicates that this is the first report of the recombinant production of active plant PPi-PFK and the characterization of its different conformations. This is a main contribution for future studies focused to better understand the enzyme properties and how it accomplishes its relevant role in plant metabolism. Fil: Muchut, Robertino José. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; Argentina Fil: Piattoni, Claudia Vanesa. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; Argentina Fil: Margarit, Ezequiel. 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: 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: 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 Fil: Iglesias, Alberto Alvaro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; Argentina |
description |
This work reports the molecular cloning and heterologous expression of the genes coding for α and β subunits of pyrophosphate-dependent phosphofructokinase (PPi-PFK) from orange. When expressed individually, both recombinant subunits were produced as highly purified monomeric proteins able to phosphorylate fructose-6-phosphate at the expenses of PPi (specific activity of 0.075 and 0.017 units. mg −1 for α and β subunits, respectively). On the other hand, co-expression rendered a α 3 β 3 hexamer with specific activity three orders of magnitude higher than the single subunits. All the conformations of the enzyme were characterized with respect to its kinetic properties and sensitivity to the regulator fructose-2,6-bisphosphate. A thorough review of current knowledge on the matter indicates that this is the first report of the recombinant production of active plant PPi-PFK and the characterization of its different conformations. This is a main contribution for future studies focused to better understand the enzyme properties and how it accomplishes its relevant role in plant metabolism. |
publishDate |
2019 |
dc.date.none.fl_str_mv |
2019-03 |
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/11336/148468 Muchut, Robertino José; Piattoni, Claudia Vanesa; Margarit, Ezequiel; Tripodi, Karina Eva Josefina; Podesta, Florencio Esteban; et al.; Heterologous expression and kinetic characterization of the α β and αβ blend of the PPi-dependent phosphofructokinase from Citrus sinensis; Elsevier Ireland; Plant Science; 280; 3-2019; 348-354 0168-9452 CONICET Digital CONICET |
url |
http://hdl.handle.net/11336/148468 |
identifier_str_mv |
Muchut, Robertino José; Piattoni, Claudia Vanesa; Margarit, Ezequiel; Tripodi, Karina Eva Josefina; Podesta, Florencio Esteban; et al.; Heterologous expression and kinetic characterization of the α β and αβ blend of the PPi-dependent phosphofructokinase from Citrus sinensis; Elsevier Ireland; Plant Science; 280; 3-2019; 348-354 0168-9452 CONICET Digital CONICET |
dc.language.none.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/abs/pii/S0168945218311075 info:eu-repo/semantics/altIdentifier/doi/10.1016/j.plantsci.2018.12.012 |
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 application/pdf application/pdf application/pdf |
dc.publisher.none.fl_str_mv |
Elsevier Ireland |
publisher.none.fl_str_mv |
Elsevier Ireland |
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_ |
1844613406072504320 |
score |
13.070432 |