Characterization and functional recovery of a novel antimicrobial peptide (CECdir-CECret) from inclusion bodies after expression in Escherichia coli

Paulina Schmitt; Luis Mercado; Mauricio Díaz; Fanny Guzmán; Gloria Arenas; Sergio H. Marshall

doi:10.1016/j.peptides.2007.12.012

Characterization and functional recovery of a novel antimicrobial peptide (CEC_dir-CEC_ret) from inclusion bodies after expression in Escherichia coli

Paulina Schmitt, Luis Mercado, Mauricio Díaz, Fanny Guzmán, Gloria Arenas, Sergio H. Marshall

INSTITUTE OF BIOLOGY

Research output: Contribution to journal › Article › peer-review

19 Scopus citations

Abstract

CEC_dir-CEC_ret is a novel non-toxic doublet 8.5 kDa peptide representing the natural coding sequence of the antimicrobial peptide Cecropin A from Drosophila melanogaster fused in-frame to its own inverted version. Expression of this cloned doublet peptide in Escherichia coli, yielded peptides that were mostly packaged into inclusion bodies. The new molecule was purified, solubilized and refolded, through a standard guanidine-based procedure. The recovered refolded peptides were then characterized by HPLC chromatography, MALDI-TOF-mass spectrometry and peptide sequencing, and finally evaluated for their antimicrobial potential. The novel doublet peptide CEC_dir-CEC_ret, displays an enhanced in vitro antimicrobial activity and action spectrum in comparison to the monomer Cecropin A.

Original language	English
Pages (from-to)	512-519
Number of pages	8
Journal	Peptides
Volume	29
Issue number	4
DOIs	https://doi.org/10.1016/j.peptides.2007.12.012
State	Published - Apr 2008

Keywords

Antibacterial peptides
Cecropin
Inclusion bodies
Refolding procedure

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10.1016/j.peptides.2007.12.012

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title = "Characterization and functional recovery of a novel antimicrobial peptide (CECdir-CECret) from inclusion bodies after expression in Escherichia coli",

abstract = "CECdir-CECret is a novel non-toxic doublet 8.5 kDa peptide representing the natural coding sequence of the antimicrobial peptide Cecropin A from Drosophila melanogaster fused in-frame to its own inverted version. Expression of this cloned doublet peptide in Escherichia coli, yielded peptides that were mostly packaged into inclusion bodies. The new molecule was purified, solubilized and refolded, through a standard guanidine-based procedure. The recovered refolded peptides were then characterized by HPLC chromatography, MALDI-TOF-mass spectrometry and peptide sequencing, and finally evaluated for their antimicrobial potential. The novel doublet peptide CECdir-CECret, displays an enhanced in vitro antimicrobial activity and action spectrum in comparison to the monomer Cecropin A.",

keywords = "Antibacterial peptides, Cecropin, Inclusion bodies, Refolding procedure",

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T1 - Characterization and functional recovery of a novel antimicrobial peptide (CECdir-CECret) from inclusion bodies after expression in Escherichia coli

AU - Schmitt, Paulina

AU - Mercado, Luis

AU - Díaz, Mauricio

AU - Guzmán, Fanny

AU - Arenas, Gloria

AU - Marshall, Sergio H.

PY - 2008/4

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AB - CECdir-CECret is a novel non-toxic doublet 8.5 kDa peptide representing the natural coding sequence of the antimicrobial peptide Cecropin A from Drosophila melanogaster fused in-frame to its own inverted version. Expression of this cloned doublet peptide in Escherichia coli, yielded peptides that were mostly packaged into inclusion bodies. The new molecule was purified, solubilized and refolded, through a standard guanidine-based procedure. The recovered refolded peptides were then characterized by HPLC chromatography, MALDI-TOF-mass spectrometry and peptide sequencing, and finally evaluated for their antimicrobial potential. The novel doublet peptide CECdir-CECret, displays an enhanced in vitro antimicrobial activity and action spectrum in comparison to the monomer Cecropin A.

KW - Antibacterial peptides

KW - Cecropin

KW - Inclusion bodies

KW - Refolding procedure

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DO - 10.1016/j.peptides.2007.12.012

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Characterization and functional recovery of a novel antimicrobial peptide (CEC_dir-CEC_ret) from inclusion bodies after expression in Escherichia coli

Abstract

Keywords

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