ABSTRACT
We investigated the ability of free fatty acids to inhibit the activity of Clostridium histolyticum collagenase (EC 3.4.24.3) and human neutrophil elastase (EC 3.4.21.37). We determined the activity of collagenase by degradation of resorufin-labeled casein fluorimetrically. The determination of the elastase activity was performed by a spectrophotometric method using a 4-nitroanilide peptide substrate. We found that most of the tested fatty acids inhibited collagenase at concentrations between 50 microM and 500 microM. For elastase we found an inhibition of the activity at concentrations between 500 nM and 50 microM. The most potent inhibitory fatty acids of both enzymes differed. Thus, as a result for collagenase we can assume that the saturated fatty acids with C(16)-C(19) were the most potent ones. For elastase the inhibition rate of unsaturated acids was much higher than the rate of the saturated ones. The highly active erucic acid with an IC(50) value of 450 nM (elastase) is remarkable.
Subject(s)
Clostridium/enzymology , Fatty Acids, Nonesterified/pharmacology , Leukocyte Elastase/antagonists & inhibitors , Microbial Collagenase/antagonists & inhibitors , Dose-Response Relationship, Drug , HumansABSTRACT
Collagenase and elastase play a critical role in the degradation of connective tissue. Aqueous and dichloromethane extracts of 15 Basidiomycetes were screened for their ability to inhibit the activity of collagenase and elastase. Collagenase (EC 3.4.24.3) was not inhibited by aqueous extracts, but by dichloromethane extracts in concentrations of 200 microg/mL. For seven extracts an IC(50) less than 200 microg/mL could be observed. Elastase (3.4.21.37) was inhibited by both aqueous and dichloromethane extracts of the Basidiomycetes, with the aqueous extracts active at concentrations of 200 microg/mL. Five extracts inhibited strongly, with an IC(50) 2-20 microg/mL. Except for four, all dichloromethane extracts inhibited the enzyme in concentrations of 2 microg/mL, nine of them very strongly at about 90%.
