FIG. 4.

Extracellular activities (A) and amount of protein (B) determined by Western blotting of subtilisin BPN′ in culture supernatants of screening host B. subtilis TEB1030 and expression host B. licheniformis H402. The protein bands correspond to a molecular mass of 28 kDa, as deduced from positions of molecular weight standards (NEB; broad range, 2 to 212 kDa, not shown) corresponding to the theoretical molecular mass of 27.6 kDa of mature BPN′. A B. subtilis strain with the vector encoding BPN′ without SP served as a control. Error bars indicate the standard deviations of proteolytic activities detected for each construct in at least three independent cultivations.