Defects in cytochrome cd1-dependent nitrite respiration of transposon Tn5-induced mutants from Pseudomonas stutzeri

Mutants with defective respiratory nitrite utilization (Nir- phenotype) were obtained by transposon Tn5 insertion into genomic DNA of the ZoBell strain of Pseudomonas stutzeri. Three representative mutants were characterized with respect to their activities of nitrite and nitric oxide reduction, cytochrome cd1 content, and pattern of soluble c-type cytochromes. Mutant strain MK201 overproduced cytochrome c552 about fourfold by comparison with the wild type, but possessed an in vitro functional cytochrome cd1. Mutant strain MK202 lacked cytochrome cd1 and, simultaneously, had low amounts of cytochrome c552 and the split alpha-peak c-type cytochrome. Strain MK203 synthesized nitrite reductase defective in the heme d1 prosthetic group. Irrespective of these biochemically distinct Nir- phenotypes, all mutants preserved the nitric oxide-reducing capability of the wild type. The mutant characteristics demonstrate that cytochrome cd1 is essential for nitrite respiration of P. stutzeri and establish the presence of a nitric oxide-reducing system distinct from cytochrome cd1. They also indicate the functional or regulatory interdependence of c-type cytochromes.