Respiratory mutants of Azotobacter vinelandii with elevated levels of cytochrome d.

A method is described for the isolation of respiratory mutants of Azotobacter vinelandii with increased amounts of d-type cytochrome by selecting for the inability to reduce tetrazolium red. Five stable mutants were obtained that had six-fold higher levels of cytochrome d, increased amounts of b-type and lower amounts of o-type and c-type cytochromes than the wild-type strain. Spectral alterations in cytochrome alpha 1 were also observed in the mutants. NADH and succinate oxidase activities of membrane particles were about two fold higher in the mutants compared to the wild-type strain. Ascorbate-N,N,N',N'-tetramethyl-p-phenylene diamine oxidase activity was barely detectable in membrane particles of the mutants. These results are consistent with an increase in the amount of the cytochrome d oxidase branch and a decrease in the amount and activity of the cytochrome o, alpha 1 oxidase branch in the mutants. Growth rates under oxygen-excess conditions and respiratory-linked proton translocation ratios of the mutant and wild-type stains were similar as were the photochemical spectral and kinetic properties of cytochrome d.