PURIFICATION AND ASSAY OF BACTERIAL COLLAGENASES

Many different types of bacteria, including pathogens, produce collagenolytic enzymes. In the past, it has been difficult to determine the role of these enzymes in host-parasite relationships because few detailed purification procedures and collagenase-specific assays have been available. The development of procedures to purify bacterial collagenases to homogeneity and assays to specifically determine their collagenolytic activity have made it possible to further study and characterize these enzymes. Assays to measure collagenolytic activity include radioactive assays to measure release of [C-14]amino acids or peptides from radioactively labeled collagen, a viscometric assay to determine decreases in viscosity of collagen solutions, ninhydrin-based assays to detect amino acids and peptides liberated from collagen, synthetic peptide assays to measure hydrolysis of synthetic peptides similar in structure to collagen, and collagen film/gel assays to detect collagen hydrolysis. This paper reviews some of the techniques available to purify and assay bacterial collagenases.