Heterologous expression, purification and refolding of an anti-listerial peptide produced by Pediococcus acidilactici K7

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-p...

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Autores principales:	Halami,Prakash M, Chandrashekar,Arun
Lenguaje:	English
Publicado:	Pontificia Universidad Católica de Valparaíso 2007
Materias:	fusion protein inclusion bodies in vitro refolding pediocin PA-1 Pediococcus acidilactici RP-HPLC
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582007000400009
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spelling	oai:scielo:S0717-345820070004000092009-01-19Heterologous expression, purification and refolding of an anti-listerial peptide produced by Pediococcus acidilactici K7Halami,Prakash MChandrashekar,Arun fusion protein inclusion bodies in vitro refolding pediocin PA-1 Pediococcus acidilactici RP-HPLC 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 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.info:eu-repo/semantics/openAccessPontificia Universidad Católica de ValparaísoElectronic Journal of Biotechnology v.10 n.4 20072007-10-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582007000400009en10.4067/S0717-34582007000400009
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	fusion protein inclusion bodies in vitro refolding pediocin PA-1 Pediococcus acidilactici RP-HPLC
spellingShingle	fusion protein inclusion bodies in vitro refolding pediocin PA-1 Pediococcus acidilactici RP-HPLC Halami,Prakash M Chandrashekar,Arun Heterologous expression, purification and refolding of an anti-listerial peptide produced by Pediococcus acidilactici K7
description	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 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.
author	Halami,Prakash M Chandrashekar,Arun
author_facet	Halami,Prakash M Chandrashekar,Arun
author_sort	Halami,Prakash M
title	Heterologous expression, purification and refolding of an anti-listerial peptide produced by Pediococcus acidilactici K7
title_short	Heterologous expression, purification and refolding of an anti-listerial peptide produced by Pediococcus acidilactici K7
title_full	Heterologous expression, purification and refolding of an anti-listerial peptide produced by Pediococcus acidilactici K7
title_fullStr	Heterologous expression, purification and refolding of an anti-listerial peptide produced by Pediococcus acidilactici K7
title_full_unstemmed	Heterologous expression, purification and refolding of an anti-listerial peptide produced by Pediococcus acidilactici K7
title_sort	heterologous expression, purification and refolding of an anti-listerial peptide produced by pediococcus acidilactici k7
publisher	Pontificia Universidad Católica de Valparaíso
publishDate	2007
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582007000400009
work_keys_str_mv	AT halamiprakashm heterologousexpressionpurificationandrefoldingofanantilisterialpeptideproducedbypediococcusacidilacticik7 AT chandrashekararun heterologousexpressionpurificationandrefoldingofanantilisterialpeptideproducedbypediococcusacidilacticik7
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Heterologous expression, purification and refolding of an anti-listerial peptide produced by Pediococcus acidilactici K7

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