Isolation of the Teichoic Acid of Bacillus Subtilis 168 by Affinity Chromatography

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Abstract
A column of insoluble concanavalin A was prepared by coupling the protein to cyanogen bromide-activated Sepharose. When autolysates of Bacillus subtilis 168 cell walls were passed over the column, the alpha glucosylated teichoic acid component of the cell wall was retained. The teichoic acid could be eluted with dilute alpha-methylglucopyranose. The teichoic acid prepared by affinity chromatography from cell wall autolysates had a higher sedimentation rate than teichoic acids obtained by conventional methods. Several authors have shown that concanavalin A (con A) forms complexes with alpha-glucosylated teichoic acids1–3. Doyle and Birdsell1 found that the teichoic acid of Bacillus subtilis 168 (trp C2) would precipitate with con A at neutral pH in dilute buffer. The formation of a precipitate was inhibited by sugars which bind to the active site of con A. This observation suggested that it should be possible to purify the teichoic acid by affinity chromatography using insoluble con A as the affinity probe. Lloyd4 and Donnelly and Goldstein5 have successfully employed insoluble con A to purify polysaccharides and glycoproteins. In this communication, we describe conditions for the rapid purification of the alpha-glucosylated teichoic acid of B. subtilis 168. The teichoic acid prepared by this procedure appears to be less degraded than teichoic acids obtained by conventional methods.