One-Step Procedure for Direct Purification of Pediocin-Like Bacteriocins and Cationic Antimicrobial Peptides from Complex Culture Medium on an Analytical, Semipreparative, and Preparative Scale. A New Type of Liquid Chromatography—Colloidal High-Performance Liquid Chromatography (CHPLC)

Abstract

A simple time- and labor-saving one-step procedure suitable for analytical-, semipreparative-, and preparative-scale preparation of bacteriocins and other cationic antibacterial peptides (for example as lysozyme) has been developed. Suspensions of bacterial cells, other colloidal- and fast-particle materials, and dissolved anionic molecules passed through the column; however, cationic bacteriocins were separated by chromatography. The purity of the target substance at more than 97% was achieved by the judgment of spectral, chromatographic, and antimicrobial activity. A many-fold concentration increase was achieved for bacteriocins, lysozyme, and bacitracin (a commercial antibiotic) in comparison with the starting solutions (see the text below). In the experiments, homemade cation-exchange columns have been used. The columns have been prepared by the grafting of glass fibers with a diameter of 0.2 μm. In this paper, applications of a new type of liquid chromatography—colloidal high-performance liquid chromatography (CHPLC)—were demonstrated.