Improved recombinant expression and purification of functional plant Rubisco.
FEBS Lett
; 593(6): 611-621, 2019 03.
Article
en En
| MEDLINE
| ID: mdl-30815863
ABSTRACT
Improving the performance of the key photosynthetic enzyme Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) by protein engineering is a critical strategy for increasing crop yields. The extensive chaperone requirement of plant Rubisco for folding and assembly has long been an impediment to this goal. Production of plant Rubisco in Escherichia coli requires the coexpression of the chloroplast chaperonin and four assembly factors. Here, we demonstrate that simultaneous expression of Rubisco and chaperones from a T7 promotor produces high levels of functional enzyme. Expressing the small subunit of Rubisco with a C-terminal hexahistidine-tag further improved assembly, resulting in a ~ 12-fold higher yield than the previously published procedure. The expression system described here provides a platform for the efficient production and engineering of plant Rubisco.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Contexto en salud:
3_ND
Problema de salud:
3_neglected_diseases
/
3_zoonosis
Asunto principal:
Ribulosa-Bifosfato Carboxilasa
/
Clonación Molecular
/
Arabidopsis
/
Chaperonas Moleculares
/
Proteínas de Unión a Fosfato
/
Proteínas de Arabidopsis
/
Chaperoninas del Grupo I
Tipo de estudio:
Prognostic_studies
Idioma:
En
Revista:
FEBS Lett
Año:
2019
Tipo del documento:
Article
País de afiliación:
Alemania