원문정보
초록
영어
Two-component oxygenases catalyze a wide variety of important oxidation reactions. Recently, we characterized a novel arylamine N-oxygenase (PrnD), a new member of the two-component oxygenase family.(1) Although arylamine N-oxygenases are widespread in nature, aminopyrrolnitrin N-oxygenase (PrnD)represents the only biochemically and mechanistically characterized arylamine N-oxygenase to date.(2) Here we report the use of bioinformatic and biochemical tools to identify and characterize the reductase component (prnF) involved in the
PrnD- catalyzed unusual arylamine oxidation. The prnF gene was identified via sequence analysis of the whole genome of Pseudomonas fluorescens and subsequently cloned and overexpressed in E. coli. The purified PrnF protein catalyzes reduction of FAD by NADH with kcat of 65 s-1 (Km = 3.2mM for FAD and 43.1mM for NADH) and supplies reduced FAD to the PrnD oxygenase component. Unlike other known reductases in two-component oxygenase
systems, PrnF strictly requires NADH as an electron donor to reduce FAD, and requires unusual protein-protein interaction with the PrnD component for the efficient transfer of reduced FAD. This PrnF enzyme represents the first cloned and characterized flavin reductase component in a novel two-component arylamine oxygenase system.