Improved techniques for the preparation of bacterial lipopolysaccharides

Abstract

Lipopolysaccharides were prepared from six organisms by the use of two cell-disruption procedures before conventional phenol–water extraction. Disruption of cells by grinding with glass beads or by digestion with hen egg white lysozyme before phenol extraction facilitated rapid purification and greater yields of lipopolysaccharide. Pretreatment of cells with lysozyme in the presence of ethylenediaminetetraacetic acid was the most efficient method in terms of lipopolysaccharide yield and ease of preparation. Increase in lipopolysaccharide yield achieved by use of the lysozyme method, compared with the conventional phenol extraction, varied from 1.7- to 12.4-fold. Preparations were designated as pure according to several criteria and were judged not to have undergone changes as a result of prephenol extraction procedures.