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To estimate the elastase activity, the supernatant (750 μl) from overnight grown (with or without SJ16 extract) cultures of P. aeruginosa (PAO1 and PAH) were incubated with elastin congo red solution (250 μl; 5 mg/ml in 0.1 M Tris–HCl pH 8;1 mM CaCl2) at 37°C for 16 h. Reaction-mixtures were centrifuged (3,000 × g for 10 min), the supernatant was collected, and the absorbance was read at 495 nm (Bjorn et al., 1979; Zhu et al., 2002).
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