In Vitro Reconstitution and Crystal Structure of P-aminobenzoate N-oxygenase (AurF) Involved in Aureothin Biosynthesis

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 2008 May 7

PMID 18458342

Citations 73

Authors

Yoo Seong Choi

Houjin Zhang

Joseph S Brunzelle

Satish K Nair

Huimin Zhao

Affiliations

Soon will be listed here.

Abstract

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.

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