3-hydroxylaminophenol mutase from Ralstonia eutropha JMP134 catalyzes a Bamberger rearrangement

3-Hydroxylaminophenol mutase from Ralstonia eutropha JMP134 is involved in the degradative pathway of 3-nitrophenol, in which it catalyzes the convers ion of 3-hydroxylaminophenol to aminohydroquinone. To show that the reactio n was really catalyzed by a single enzyme without the release of intermedia tes, the corresponding protein was purified to apparent homogeneity from an extract of cells grown on 3-nitrophenol as the nitrogen source and succina te as the carbon and energy source. 3-Hydroxylaminophenol mutase appears to be a relatively hydrophobic but soluble and colorless protein consisting o f a single 62-kDa polypeptide. The pi was determined to be at pH 4.5. In a database search, the NH2-terminal amino acid sequence of the undigested pro tein and of two internal sequences of 3-hydroxylaminophenol mutase were fou nd to be most similar to those of glutamine synthetases from different spec ies. Hydroxylaminobenzene, 4-hydroxylaminotoluene, and 2-chloro-5-hydroxyla minophenol, but not 4-hydroxylaminobenzoate, can also serve as substrates f or the enzyme. The enzyme requires no oxygen or added cofactors for its rea ction, which suggests an enzymatic mechanism analogous to the acid-catalyze d Bamberger rearrangement.