para-Hydroxybenzoate supported nitrogen fixation in Azotobacter vinelandii strain OP (13705)

Azotobacter vinelandii cells grew with molecular nitrogen and p-hydroxybenzoate as the sole added nitrogen and carbon sources. Nitrogenase activity in p-hydroxybenzoate grown cells was demonstrated with the acetylene reduction assay. Cell extracts contained the enzymes p-hydroxybenzoate hydroxylase (EC 1.14.13.2) and protocatechuate 3,4-dioxygenase (EC 1.13.1.3); oxygenases associated with p-hydroxybenzoate metabolism. These enzymes separated from respiration particles with gel filtration chromatography, indicating that they are soluble and not membrane bound. This evidence indicates that oxygen enters to the inner face of the cytoplasmic membrane during nitrogen fixation.