Quinaldine 4-oxidase from Arthrohacter sp. Rub1 a, an inducible molybdenum-containing hydroxylase, was purified to homogeneity by an optimized five-step procedure. Molecular oxygen is proposed as physiological electron acceptor. Electrons are also transferred to artificial electron acceptors with Eh > -8 mV. The molybdo-ironhlfur flavoprotein regiospecifically attacks its N-heterocyclic substrates :isoquinoline and phthalazine are hydroxylated adjacent to the N-heteroatom at C1, whereas quinaldine, quinoline, cinnoline and quinazoline are hydroxylated at C4. Additionally, the aromatic aldehydes benzaldehyde, salicylaldehyde, vanillin and cinnamaldehyde are oxidized to the corresponding carboxylic acids, whereas short-chain aliphatic aldehydes are not.Quinaldine 4-oxidase is compared to the two molybdenum-containing hydroxylases quinoline 2-oxidoreductase from Pseudomonas putida 86 [Tshisuaka, B., Kappl, R., Hiittermann, J. with respect to the substrates converted and the electron-acceptor specificities. These dehydrogenases hydroxylate their N-heterocyclic substrates exclusively adjacent to the heteroatom. Whereas the aldehydes tested are scarcely oxidized by quinoline 2-oxidoreductase, isoquinoline 1 -0xidoreductase catalyzes the oxidation of the aromatic aldehydes, although being progressively inhibited. Neither quinoline 2-oxidoreductase nor isoquinoline 1 -0xidoreductase transfer electrons to oxygen. Otherwise, the spectrum of electron acceptors used by quinoline 2-oxidoreductase and quinaldine 4-oxidase is identical. However, isoquinoline I-oxidoreductase differs in its electron-acceptor specificity.Quinaldine 4-oxidase is unusual in its substrate and electron-acceptor specificity. This enzyme is able to function as oxidase or dehydrogenase, it oxidizes aldehydes, and it catalyzes the nucleophilic attack of N-containing heterocyclic compounds at two varying positions depending on the substrate.Keywords: benzodiazines ; benzopyridine derivatives ; aldehyde oxidation ; molybdenum-containing hydroxylase ; quinaldine 4-oxidase.Quinaldine 4-oxidoreductase from the gram-positive bacterium Arthrobacter sp. Rii61 a, an inducible molybdenum-containing hydroxylase, catalyzes the initial step in quinaldine (2-methylquinoline) degradation (Hund et al., 1990) : quinaldine is converted to 1 H-4-oxoquinaldine with concomitant reduction of a suitable electron acceptor. The oxygen atom incorporated into the product derives from water. Quinaldine 4-oxidoreductase
Abbreviations. INT, 2-(4-iodophenyl)-3-(4-nitrophenyl)-5-phenyl-2H-tetrazolium chloride; 11 OR, isoquinoline l-oxidoreductase; Q20R, quinoline 2-oxidoreductase; NBT, 2,2'-di-p-nitrophenyl-5,5'-diphenyl-3,3'-~3,3'-dimethoxy-4,4'-diphenylene]-dite~azolium chloride.Enzymes.