Rapid two-step procedure for large-scale purification of pediocin-like bacteriocins and other cationic antimicrobial peptides from complex culture medium.
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.
Uteng
Marianne
M
Department of Biochemistry, University of Oslo, Blindern, 0316 Oslo, Norway.
Hauge
Håvard Hildeng
HH
Brondz
Ilia
I
Nissen-Meyer
Jon
J
Fimland
Gunnar
G
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*
D023181:Antimicrobial Cationic Peptides; D001430:Bacteriocins; D017077:Culture Media, Conditioned