Artículo
Heterologous expression and kinetic characterization of the α β and αβ blend of the PPi-dependent phosphofructokinase from Citrus sinensis
Muchut, Robertino José
; Piattoni, Claudia Vanesa
; Margarit, Ezequiel
; Tripodi, Karina Eva Josefina
; Podesta, Florencio Esteban
; Iglesias, Alberto Alvaro
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Fecha de publicación:
03/2019
Editorial:
Elsevier Ireland
Revista:
Plant Science
ISSN:
0168-9452
Idioma:
Inglés
Tipo de recurso:
Artículo publicado
Clasificación temática:
Resumen
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.
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Articulos(CEFOBI)
Articulos de CENTRO DE EST.FOTOSINTETICOS Y BIOQUIMICOS (I)
Articulos de CENTRO DE EST.FOTOSINTETICOS Y BIOQUIMICOS (I)
Articulos(IAL)
Articulos de INSTITUTO DE AGROBIOTECNOLOGIA DEL LITORAL
Articulos de INSTITUTO DE AGROBIOTECNOLOGIA DEL LITORAL
Citación
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
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