FIG. 3.

High-cell-density cultivation of B. subtilis TEB1030 carrying BPN′ fused to SPs wtSP and dBli00338. Cells were grown in fed-batch mode in a fermentor (culture volume: 3 liters), and culture supernatants were analyzed for extracellular protease activity (A) and amount of extracellular protease protein (B and C) by SDS-PAGE and subsequent staining with Coomassie brilliant blue (B) and Western blotting using antibodies against subtilisin BPN′ (C). The protein bands correspond to a molecular mass of 28 kDa, as deduced from positions of molecular weight standards (New England Biolabs [NEB]; broad range, 2 to 212 kDa, not shown) and thus correspond to the theoretical molecular mass of 27.6 kDa of mature BPN′.