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. 2018 Feb 1;9:105. doi: 10.3389/fmicb.2018.00105

FIGURE 5.

FIGURE 5

ED367 heterologous ComX biological activity decay (A) and the purification of heterologous ComX on HPLC (B). (A) The ComX biological activity decay of the E. coli ED367 spent medium containing heterologously expressed ComX, using a signal deficient biosensor strain BM1456 with a PsrfAA-yfp fluorescent reporter fusion. The spent medium was incubated for 24 h at 37°C with and without subtilisin. When adding subtilisin, the ComX biological activity decay increases. Averages and SEM of three independent biological replicates are shown. A Mann–Whitney U-test was performed to determine statistical significance of discussed measurements (∗p < 0.05). (B) Chromatograms of the E. coli ED367 M9 spent media that were incubated for 24 h at 37°C with and without subtilisin. After treating the medium with subtilisin, the two peaks containing ComX are no longer present in the medium that contained heterologously expressed ComX. As a control a parallel experiments were made where non-IPTG induced E. coli ED367 spent medium was incubated for 24 h at 37°C with and without subtilisin.