Fig. 2

Secretion and activation of BLP by B. subtilis. B. subtilis strain 168 was transformed with pHY300PLK (empty vector), pHY-BLP01, pHY-BLP02, or pHY-BLP03 for BLP expression. Cells were grown at 30 °C for 72 h in modified 2 × L-Mal medium. A SDS-PAGE analysis of culture supernatants. The supernatants were diluted two-fold with SDS reducing buffer. Five microliters of each sample were applied to SDS-PAGE. The expression plasmids are indicated above the image. The arrow indicates the position of mature BLP. B Pentaglycine cleavage activity assay of culture supernatants using the pentaglycine-containing FRET substrate FRET-GGGGG. C Bactericidal assay of the purified BLP against S. aureus NCTC8325. The bactericidal activity was measured at a final protein concentration of 1.5 µg/mL. The results presented are the means of three individual experiments. Error bars represent the standard errors of the means