title: Rapid two-step procedure for large-scale purification of pediocin-like bacteriocins and other cationic antimicrobial peptides from complex culture medium.
abstract: A rapid and simple two-step procedure suitable for both small- and large-scale purification of pediocin-like bacteriocins and other cationic peptides has been developed. In the first step, the bacterial culture was applied directly on a cation-exchange column (1-ml cation exchanger per 100-ml cell culture). Bacteria and anionic compounds passed through the column, and cationic bacteriocins were subsequently eluted with 1 M NaCl. In the second step, the bacteriocin fraction was applied on a low-pressure, reverse-phase column and the bacteriocins were detected as major optical density peaks upon elution with propanol. More than 80% of the activity that was initially in the culture supernatant was recovered in both purification steps, and the final bacteriocin preparation was more than 90% pure as judged by analytical reverse-phase chromatography and capillary electrophoresis.
authors.affiliation: Department of Biochemistry, University of Oslo, Blindern, 0316 Oslo, Norway.
authors.forename: Marianne
authors.initials: M
authors.lastname: Uteng
authors.forename: Håvard Hildeng
authors.initials: HH
authors.lastname: Hauge
authors.forename: Ilia
authors.initials: I
authors.lastname: Brondz
authors.forename: Jon
authors.initials: J
authors.lastname: Nissen-Meyer
authors.forename: Gunnar
authors.initials: G
authors.lastname: Fimland
mesh_terms: D023181:Antimicrobial Cationic Peptides / Q000302:isolation & purification*; D001430:Bacteriocins / Q000302:isolation & purification*; D001709:Biotechnology / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000379:methods; D017077:Culture Media, Conditioned / Q000737:chemistry; D010376:Pediococcus / Q000254:growth & development / Q000378:metabolism*
chemical_list: D023181:Antimicrobial Cationic Peptides; D001430:Bacteriocins; D017077:Culture Media, Conditioned