Heterologous expression, purification and refolding of an anti-listerial peptide produced by Pediococcus acidilactici K7
Electron. j. biotechnol
;
10(4): 563-569, oct. 2007. ilus, graf, tab
Artículo
en Inglés
| LILACS
| ID: lil-504119
ABSTRACT
The fusion protein, 6XHis-Xpress-PedA was constructed and expressed in Escherichia coli BL21 (DE3). The presence of a 12.8 kDa recombinant protein, localized in inclusion bodies (IBs) at high concentration, was confirmed by SDS-PAGE analysis and by western blotting using anti-His antibody. The rec-pediocin was purified by Nickel-nitrilotriacetic acid beads and refolded using 5 mM of beta-mercaptoethanol along with 1 M glycine. Results indicated that the refolded rec-pediocin had an early elution profile in the RP-HPLC when compared to the unfolded protein and it exhibited biological activity against Listeria monocytogenes V7 which was approximately 25 times less active compared to native counterpart. The final yield of purified rec-pediocin was 3 mg/l of the culture and is estimated to be 8-10 times higher than the purification by conventional methods.
Texto completo:
Disponible
Índice:
LILACS (Américas)
Asunto principal:
Pediococcus
/
Bacteriocinas
/
Proteínas Recombinantes de Fusión
/
Cuerpos de Inclusión
/
Cromatografía Líquida de Alta Presión
Idioma:
Inglés
Revista:
Electron. j. biotechnol
Asunto de la revista:
Biotecnologia
Año:
2007
Tipo del documento:
Artículo
País de afiliación:
India
Institución/País de afiliación:
Central Food Technological Research Institute/IN
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