Main content area

In vitro reconstitution and crystal structure of p-aminobenzoate N-oxygenase (AurF) involved in aureothin biosynthesis

Choi, Yoo Seong, Zhang, Houjin, Brunzelle, Joseph S., Nair, Satish K., Zhao, Huimin
Proceedings of the National Academy of Sciences of the United States of America 2008 v.105 no.19 pp. 6858-6863
Streptomyces thioluteus, antibiotics, biosynthesis, crystal structure, enzyme activity, hydroxylamine, nitrobenzoic acids, oxidation, p-aminobenzoic acid, polyketide synthases
p-aminobenzoate N-oxygenase (AurF) from Streptomyces thioluteus catalyzes the formation of unusual polyketide synthase starter unit p-nitrobenzoic acid (pNBA) from p-aminobenzoic acid (pABA) in the biosynthesis of antibiotic aureothin. AurF is a metalloenzyme, but its native enzymatic activity has not been demonstrated in vitro, and its catalytic mechanism is unclear. In addition, the nature of the cofactor remains a controversy. Here, we report the in vitro reconstitution of the AurF enzyme activity, the crystal structure of AurF in the oxidized state, and the cocrystal structure of AurF with its product pNBA. Our combined biochemical and structural analysis unequivocally indicates that AurF is a non-heme di-iron monooxygenase that catalyzes sequential oxidation of aminoarenes to nitroarenes via hydroxylamine and nitroso intermediates.