Assays for Studying Pseudomonas aeruginosa Secreted Proteases

Proteolytic activity plays an essential role in Pseudomonas aeruginosa adaptation and survival in challenging environments, including the infection site. Here, a short review of the eight known proteases secreted by P. aeruginosa and of the methods used to detect their activity is provided. In addition, three simple and handy methods routinely used in our laboratory to detect proteases are described in detail. In particular, the skim milk plate assay and the azocasein assay are useful for the detection of whole proteases activity in colony-growing and cell-free culture supernatants, respectively. Conversely, the Elastin Congo-red assay allows detecting the activity of the LasB elastase, the major protease secreted by P. aeruginosa, in cell-free culture supernatants.