Data/hora: 01/06/2024 10:49:19

Provedor de dados: | 69
País: | Chile
Título: | Intein-mediated expression of cecropin in Escherichia coli
Autores: | Díaz,Mauricio; Venturini,Elena; Marchetti,Stefano; Arenas,Gloria; Marshall,Sergio H.
Data: | 2012-03-01
Ano: | 2012
Palavras-chave: | Antimicrobial; Cecropin; Fusion; Intein; Peptide; Soluble.
Resumo: | Different strategies have been used to overcome the difficulties to produce antimicrobial peptides. Here we used Intein Mediated Purification with an Affinity Chitin-binding Tag (IMPACT-System, New England Biolabs) for the expression of the antimicrobial peptide cecropin to reduce its sensitivity to intracellular proteases and use its inducible self-cleaving capability to remove the carrier. Cecropin was cloned into suitable expression vector pTYB11, and expression induced by IPTG in Escherichia coli ER2566. The use of 22ºC induction allowed the expression of cecropin with its intein carrier in soluble form. Cell extracts were purified by chitin affinity chromatography and intein-mediated splicing of the target protein was achieved by thiol addition, obtaining a final yield of 2.5 mg cecropin/l. Cecropin cleaved from the intein had its proper biologically active form, showing a micromolar antimicrobial activity against Vibrio ordalii, Vibrio alginolyticus and Escherichia coli.
Tipo: | Journal article
Idioma: | Inglês
Identificador: | http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582012000200003
Editor: | Pontificia Universidad Católica de Valparaíso
Formato: | text/html
Fonte: | Electronic Journal of Biotechnology  v.15 n.2 2012