Falgpa

Collagenase clostridium histolyticum (Sigma Aldrich) was used for this experiment, and its activity was determined with the aid of a spectrophotometer using N-[3-(2-furyl)acryloyl]-Leu-Gly-Pro-Ala (FALGPA; Sigma Aldrich) as the substrate in accordance with the protocol of Wittenauer et al. [51 (link)]. The decrease in the absorbance of the FALGPA was followed for 20 min at 335 nm using a microplate reader (BioTek ELX800; BioTek Instruments, Colmar, France). The measurements were conducted in triplicate, and the anti-collagenase activity was revealed as the percent inhibition relative to the control (extraction solvent) for every extract sample. 1,10-Phenantroline (100 µM) was used as the specific inhibitor of collagenase.

The collagenase assay was performed according to Wittenauer et al. (2015) (link). Collagenase from Clostridium histolyticum (Sigma Aldrich) was used with the substrate N-[3-(2-furyl)acryloyl]-Leu-Gly-Pro-Ala (FALGPA; Sigma Aldrich) and the decrease in absorbance of FALGPA was monitored at 335 nm over a period of 20 min, using a BioTek ELX800 absorbance microplate reader (BioTek Instruments, Colmar, France). All the reactions were performed in triplicate, and the anti-collagenase activity was detected as a percentage of inhibition relative to the control (by adding the same volume of extraction solvent) for each extract. 1,10-Phenantroline (100 µM) was used as the specific inhibitor of collagenase leading to an inhibition of 33.6 ± 2.2%.