title: Recombinant expression and purification of the antimicrobial peptide magainin-2.
abstract: Magainin-2 (MAG2) is a polycationic antimicrobial peptide isolated from the skin of the African clawed frog Xenopus laevis. It has a wide spectrum of antimicrobial activities against gram-positive and gram-negative bacteria, fungi, and induces osmotic lysis of protozoa. MAG2 also possesses antiviral and antitumoral properties. These activities make this peptide a promising candidate for therapeutic applications. Recombinant expression systems are necessary for the affordable production of large amounts of the biologically active peptide. In this work, MAG2 has been cloned to the N-terminal of a family III carbohydrate-binding module fused to the linker sequence (LK-CBM3) from Clostridium thermocellum; a formic acid recognition site was introduced between the two modules for chemical cleavage of the peptide. The recombinant protein MAG2-LK-CBM3 was expressed in Escherichia coli BL21 (DE3) and MAG2 was successfully cleaved and purified from the fusion partner LK-CBM3. Its functionality was confirmed by testing its activity against gram-negative bacteria.
authors.affiliation: Institute for Biotechnology and Bioengineering (IBB), Centre of Biological Engineering, Universidade do Minho, Campus de Gualtar, 4710-057 Braga, Portugal.
authors.forename: Reinaldo
authors.initials: R
authors.lastname: Ramos
authors.forename: Susana
authors.initials: S
authors.lastname: Moreira
authors.forename: Ana
authors.initials: A
authors.lastname: Rodrigues
authors.forename: Miguel
authors.initials: M
authors.lastname: Gama
authors.forename: Lucília
authors.initials: L
authors.lastname: Domingues
mesh_terms: D000818:Animals; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D054806:Magainins / Q000235:genetics* / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification; D029867:Xenopus Proteins / Q000235:genetics* / Q000302:isolation & purification*; D014982:Xenopus laevis / Q000235:genetics / Q000378:metabolism
chemical_list: D054806:Magainins; D011994:Recombinant Proteins; D029867:Xenopus Proteins; C053301:magainin 2 peptide, Xenopus