title: Functionality of biphenyl 2,3-dioxygenase components in naphthalene 1,2-dioxygenase.
abstract: Naphthalene 1,2-dioxygenase (Nap dox) and biphenyl 2,3-dioxygenase (Bph dox) are related enzymes that have differentiated during evolution as their specificity has changed. Although their component arrangement is similar, the structure of each component has been modified quite extensively. The purpose of this work was to determine the catalytic capacity of purified Nap dox toward chlorobiphenyls and to investigate the functionality of Bph dox components in the Nap dox system. Both enzyme systems were purified by affinity chromatography as histidine-tagged fused proteins. Data show for the first time that Nap dox can catalyze the oxygenation of all three monochlorobiphenyl isomers, but it is unable to hydroxylate 2,5-, 2,2'-, 3,3'-, 4,4'-di- and 2,2',5,5'-tetrachlorobiphenyl. The rates of cytochrome c reduction by the ferredoxin components of the two enzymes were identical when the Bph dox reductase component was used in the assay, showing an efficient electron transfer between the Bph dox reductase component and the Nap dox ferredoxin. However, when the Bph dox ferredoxin was used to reconstitute a hybrid Nap dox, the enzyme was only 22% as active as the parental enzyme. These data are discussed in terms of the potential use of Nap dox for the development of enhanced chlorobiphenyl-degrading dioxygenases.
authors.affiliation: INRS-Santé, Université du Québec, Canada.
authors.forename: D
authors.initials: D
authors.lastname: Barriault
authors.forename: M
authors.initials: M
authors.lastname: Sylvestre
mesh_terms: D049308:Dioxygenases; D004926:Escherichia coli; D009097:Multienzyme Complexes / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D010105:Oxygenases / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D016133:Polymerase Chain Reaction; D013329:Structure-Activity Relationship
chemical_list: D009097:Multienzyme Complexes; D010105:Oxygenases; D049308:Dioxygenases; C092087:biphenyl-1,2-dioxygenase; C038703:naphthalene dioxygenase