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Electronic Journal of Biotechnology
Universidad Católica de Valparaíso
ISSN: 0717-3458
Vol. 10, No. 4, 2007, pp. 563-569
Bioline Code: ej07055
Full paper language: English
Document type: Research Article
Document available free of charge

Electronic Journal of Biotechnology, Vol. 10, No. 4, 2007, pp. 563-569

 en Heterologous expression, purification and refolding of an anti-listerial peptide produced by Pediococcus acidilactici check for this species in other resources K7
Halami, Prakash M. & Chandrashekar, Arun

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 β-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 check for this species in other resources 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.

Keywords
fusion protein, inclusion bodies, in vitro refolding, pediocin PA-1, Pediococcus acidilactici, RP-HPLC.

 
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