Extracellular protease detectionwas done
after staining agar plateswith Coomassie blue (0.25%, w/v) for 1 h in
methanolacetic acid-water 5:1:4 (v/v/v) and destaining with methanolacetic
acid-water. Regions of enzyme activity were detected as
clear areas, indicating that hydrolysis of the substrates had occurred.