Effects of gentamicin on trypsin, chymotrypsin, and collagenase

The effects of gentamicin on three proteolytic enzymes were studied. Gentamicin was tested at concentrations of 0.5-500 microgram/ml. Trypsin was tested at 0.5 microgram/ml using p-tosyl-L-arginine methyl ester and at 0.1 and 0.5 microgram/ml using azocoll as the substrate. Chymotrypsin was tested at 0.1 and 0.5 microgram/ml with azocoll. A soluble [14C]collagen assay was used to measure activity of collagenase derived from Clostridium histolyticum. The profiles of proteolytic activity vs. gentamicin concentration were similar for all three enzymes. At lower concentrations of gentamicin (less than 70 microgram/ml), there were two peaks of enhanced protease activity generally followed by inhibition. These unusual multiphasic effects of gentamicin on three different proteases are not presently understood, but they imply a previously unreported mode of action for this antibiotic.