The concentrations of PEP were measured fluorometrically according to the kits’ manufacturer’s guidelines (MAK102, Sigma-Aldrich) (40). P. mirabilis was homogenized in 3 N HClO4 on ice in a 1:5 ratio (mg wet mass: μl of 3 N HClO4). The samples were neutralized with 3 M KHCO3 to achieve pH 7.4 and centrifuged (13,000g, 4°C, 5 min). Three milliliters of the resulting supernatants was incubated in 100 ml of master mix for 30 min and 1 hour for the measurement of the PEP concentrations. Last, the fluorescence intensity (excitation, 535 nm; emission, 587 nm) of the samples was measured. The concentrations of PEP were calculated by building the calibration curve for 0 to 0.5 nmol standards.

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