Hydroxylation reaction catalyzed by the Burkholderia cepacia AC1100 bacterial strain. Involvement of the chlorophenol-4-monooxygenase.
The Burkholderia cepacia AC1100 strain, known to degrade the herbicide, 2,4,5-Trichlorophenoxyacetic acid (2,4,5-T), is able to metabolize 4-hydroxyarylaldehyde, not only into the corresponding acid, but also into a new hydroquinone, 2,5-dihydroxyarylaldehyde. When incubated with resting AC1100 cells or cell-free extracts, syringaldehyde and 3,5-dimethoxy-4-hydroxybenzaldehyde were converted into such metabolites, identified by comparison of their mass and 1H-NMR spectra with those of authentic chemically synthesized samples. With 5-bromovanillin, only one metabolite was formed, the structure of which was identified as 2, 5-dihydroxy-4-methoxy-6-bromobenzaldehyde through 1H-NMR two-dimensional NOESY experiments. All these products result formally from a para hydroxylation of the phenol followed by the cis migration of the aldehyde. This reaction is the only one to be associated with the 2,4,5-T degradation pathway, as the acid formation was retained when the AC1100 strain had lost its degradation ability. Through competitive experiments with halophenols and methimazole, an alternative substrate of flavin monooxygenase, the chlorophenol-4-monooxygenase was recognized to be the enzyme involved in the hydroxylation of 4-hydroxyarylaldehyde. The purified enzyme, previously reported to catalyze the para hydroxylation or dehalogenating hydroxylation of chlorophenols, also promotes this hydroxylation reaction in the presence of NADH and FAD. The kcat value determined for the best substrate, syringaldehyde, 0. 08 s-1, was about 20% of that obtained for 2,6-dichlorophenol hydroxylation (0.38 s-1).
Martin
G
G
Laboratoire de Chimie et Biochimie Pharmacologiques et Toxicologiques, URA 400 CNRS, Univesité René Descartes, Paris, France.
Dijols
S
S
Capeillere-Blandin
C
C
Artaud
I
I
D000447:Aldehydes / Q000378:metabolism; D001547:Benzaldehydes / Q000378:metabolism; D001667:Binding, Competitive; D016956:Burkholderia cepacia / Q000201:enzymology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005182:Flavin-Adenine Dinucleotide / Q000378:metabolism; D006900:Hydroxylation; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D006899:Mixed Function Oxygenases / Q000378:metabolism*; D015394:Molecular Structure; D009243:NAD / Q000378:metabolism; D013379:Substrate Specificity
D000447:Aldehydes; D001547:Benzaldehydes; D004791:Enzyme Inhibitors; D009243:NAD; D005182:Flavin-Adenine Dinucleotide; C069665:syringaldehyde; D006899:Mixed Function Oxygenases; C099366:chlorophenol 4-monooxygenase