Proteinase affinity chromatography on bacitracin-Sepharose.

Antibiotic-cyclopeptide bacitracin covalently bound to Sepharose proved to be an efficient general ligand for affinity chromatography of aspartyl, serine, and metalloproteinases from various sources. The yields of purified enzymes varied from 50 to 180%. New experimental data extend the application of bacitracin-Sepharose for affinity chromatography of cysteine proteinases--papain, bromelain, and ficin. Hence, bacitracin acts as a ligand which more or less efficiently binds proteinases that belong to all the main classes of these enzymes. Bacitracin, being a weak proteinase inhibitor of broad specificity, interacts with the substrate-binding sites of proteinases, which explains its efficiency as a ligand.