MODIFIED FALGPA ASSAY FOR CELL-ASSOCIATED COLLAGENOLYTIC ACTIVITY

A continuous spectrophotometric assay monitoring the hydrolysis of the synthetic peptide 2-furanacryloyl-L-leucylglycyl-L-prolyl-alanine (FALGPA) is used to measure collagenase activity of both bacterial and vertebral collagenases. In the present study, a protocol was developed to adapt this assay to the measurement of cell-associated FALGPA hydrolytic activity in bacteria. The bacteria tested included Bacillus cereus, Streptococcus agalactiae, Streptococcus mutans, Enterococcus faecalis, and Escherichia coli, and various levels of activity were identified. The method presented here allows the detection of FALGPA hydrolysis using a small quantity of cells without the need for prior purification of the collagenolytic enzyme or collection and concentration of a large volume of culture supernatant fluid.