FIG. 3.

NosZ is expressed in P. putida and translocated to the periplasm. (A) Cell extracts from growing cultures of P. putida strains RZ and RE were analyzed by Western blotting over an 18-h period. Coexpression of tatE in strain RE resulted in complete translocation of NosZ. (B) Western blot analysis of periplasm (PP) and cytoplasm (CP). RZ, RY, RL, and RE are the different nos gene combinations expressed in P. putida DSM50906 (see Fig. 1). wt, nontransformed wild type; P. st., processed NosZ protein from P. stutzeri MK21. For the conditions used for cell growth, cell fractionation, preparation of cell extract, and immunoblotting see Materials and Methods.