A bacterium having collagenolytic activity was isolated from soil and identified as Bacillus licheniformis. The collagenolytic enzyme was obtained in a medium containing gelatin and purified to apparent homogeneity on the basis of SDS-polyacrylamide gel electrophoresis. The purified enzyme had a molecular weight of approximately 29,000, and hydrolyzed native, insoluble collagen. In addition, the enzyme hydrolyzed soluble collagen, gelatin, hemoglobin and casein, but not synthetic oligopeptides, usually used as substrates for collagenase assay.