title: Nonmicrobial Nitrophenol Degradation via Peroxygenase Activity of Dehaloperoxidase-Hemoglobin from Amphitrite ornata.
abstract: The marine hemoglobin dehaloperoxidase (DHP) from Amphitrite ornata was found to catalyze the H2O2-dependent oxidation of nitrophenols, an unprecedented nonmicrobial degradation pathway for nitrophenols by a hemoglobin. Using 4-nitrophenol (4-NP) as a representative substrate, the major monooxygenated product was 4-nitrocatechol (4-NC). Isotope labeling studies confirmed that the O atom incorporated was derived exclusively from H2O2, indicative of a peroxygenase mechanism for 4-NP oxidation. Accordingly, X-ray crystal structures of 4-NP (1.87 Å) and 4-NC (1.98 Å) bound to DHP revealed a binding site in close proximity to the heme cofactor. Peroxygenase activity could be initiated from either the ferric or oxyferrous states with equivalent substrate conversion and product distribution. The 4-NC product was itself a peroxidase substrate for DHP, leading to the secondary products 5-nitrobenzene-triol and hydroxy-5-nitro-1,2-benzoquinone. DHP was able to react with 2,4-dinitrophenol (2,4-DNP) but was unreactive against 2,4,6-trinitrophenol (2,4,6-TNP). pH dependence studies demonstrated increased reactivity at lower pH for both 4-NP and 2,4-DNP, suggestive of a pH effect that precludes the reaction with 2,4,6-TNP at or near physiological conditions. Stopped-flow UV-visible spectroscopic studies strongly implicate a role for Compound I in the mechanism of 4-NP oxidation. The results demonstrate that there may be a much larger number of nonmicrobial enzymes that are underrepresented when it comes to understanding the degradation of persistent organic pollutants such as nitrophenols in the environment.
authors.affiliation: Department of Chemistry, North Carolina State University , Raleigh, North Carolina , 27695-8204.
authors.forename: Nikolette L
authors.initials: NL
authors.lastname: McCombs
authors.affiliation: Department of Chemistry, North Carolina State University , Raleigh, North Carolina , 27695-8204.
authors.forename: Jennifer
authors.initials: J
authors.lastname: D'Antonio
authors.affiliation: Department of Chemistry, North Carolina State University , Raleigh, North Carolina , 27695-8204.
authors.forename: David A
authors.initials: DA
authors.lastname: Barrios
authors.affiliation: Department of Chemistry, North Carolina State University , Raleigh, North Carolina , 27695-8204.
authors.forename: Leiah M
authors.initials: LM
authors.lastname: Carey
authors.affiliation: Department of Chemistry, North Carolina State University , Raleigh, North Carolina , 27695-8204.
authors.forename: Reza A
authors.initials: RA
authors.lastname: Ghiladi
mesh_terms: D000818:Animals; D002384:Catalysis; D006454:Hemoglobins / Q000378:metabolism*; D006861:Hydrogen Peroxide / Q000378:metabolism; D006899:Mixed Function Oxygenases / Q000378:metabolism*; D009596:Nitrophenols; D010084:Oxidation-Reduction; D010100:Oxygen / Q000378:metabolism*; D010544:Peroxidases / Q000378:metabolism*; D011077:Polychaeta / Q000201:enzymology*
chemical_list: D006454:Hemoglobins; D009596:Nitrophenols; D006861:Hydrogen Peroxide; D006899:Mixed Function Oxygenases; C098661:DHP I dehaloperoxidase; D010544:Peroxidases; C022333:peroxygenase; D010100:Oxygen; C024836:4-nitrophenol