2.4. Proteolytic activity
Proteolysis was assayed against gelatin on agar plates, using a
modified method (Vermelho et al., 1996). Briefly, a loopful of the
culture to be evaluated was inoculated on agar plates with the
detection medium (2% (w/v) sucrose (Merck), 0.5% (w/v) yeast
extract (Britania), 2% (w/v) peptone (Britania), 1.5% (w/v) agar (Bri-
tania) autoclaved and supplemented with 1% (w/v) gelatin) and
incubated at 37 C for 48 h. Extracellular protease detectionwas done
after staining agar plates with Coomassie blue (0.25%, w/v) for 1 h in
methanolacetic acid-water 5:1:4 (v/v/v) and destaining with meth-
anolacetic acid-water. Regions of enzyme activity were detected as
clear areas, indicating that hydrolysis of the substrates had occurred.
2.4. Proteolytic activityProteolysis was assayed against gelatin on agar plates, using amodified method (Vermelho et al., 1996). Briefly, a loopful of theculture to be evaluated was inoculated on agar plates with thedetection medium (2% (w/v) sucrose (Merck), 0.5% (w/v) yeastextract (Britania), 2% (w/v) peptone (Britania), 1.5% (w/v) agar (Bri-tania) autoclaved and supplemented with 1% (w/v) gelatin) andincubated at 37 C for 48 h. Extracellular protease detectionwas doneafter staining agar plates with Coomassie blue (0.25%, w/v) for 1 h inmethanolacetic acid-water 5:1:4 (v/v/v) and destaining with meth-anolacetic acid-water. Regions of enzyme activity were detected asclear areas, indicating that hydrolysis of the substrates had occurred.
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