title: Crystallization and diffraction data of 1H-3-hydroxy-4-oxoquinoline 2,4-dioxygenase: a cofactor-free oxygenase of the alpha/beta-hydrolase family.
abstract: 1H-3-Hydroxy-4-oxoquinoline 2,4-dioxygenase (QDO) from Pseudomonas putida 33/1 catalyses the oxygenolysis of 1H-3-hydroxy-4-oxoquinoline to form N-formylanthranilic acid and carbon monoxide without the aid of cofactors. Both N-terminally His6-tagged and native QDO were overexpressed in Escherichia coli and purified by conventional chromatographic procedures. Untagged QDO, but not His6-tagged QDO, was crystallized by the vapour-diffusion method, giving hexagonal bipyramid crystals belonging to space group P6(1)22. Selenomethionine-containing native QDO was prepared and crystallized under identical conditions. The unit-cell parameters were a = b = 90.1, c = 168.6 A, alpha = beta = 90, gamma = 120 degrees. Using synchrotron radiation, these crystals diffract to 2.5 A. The expression, purification and crystallization of QDO are reported here.
authors.affiliation: Research School of Chemistry, Australian National University, Acton, ACT 0200, Australia.
authors.forename: Ruhu
authors.initials: R
authors.lastname: Qi
authors.forename: Susanne
authors.initials: S
authors.lastname: Fetzner
authors.forename: Aaron J
authors.initials: AJ
authors.lastname: Oakley
mesh_terms: D001173:Arthrobacter / Q000201:enzymology; D001483:Base Sequence; D003460:Crystallization; D018360:Crystallography, X-Ray; D017931:DNA Primers; D049308:Dioxygenases / Q000737:chemistry* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D006867:Hydrolases / Q000737:chemistry*; D011487:Protein Conformation; D016958:Pseudomonas putida / Q000201:enzymology; D013379:Substrate Specificity
chemical_list: D017931:DNA Primers; C101932:1H-3-hydroxy-4-oxoquinaldine 2,4-dioxygenase (decyclizing, CO-forming); D049308:Dioxygenases; D006867:Hydrolases