Cloning, sequence analysis, and expression of the Pseudomonas putida 33/1 1H-3-hydroxy-4-oxoquinoline 2,4-dioxygenase gene, encoding a carbon monoxide forming dioxygenase.
1H-3-hydroxy-4-oxoquinoline 2,4-dioxygenase (Qdo) from the 1H-4-oxoquinoline utilizing Pseudomonas putida strain 33/1, which catalyzes the cleavage of 1H-3-hydroxy-4-oxoquinoline to carbon monoxide and N-formylanthranilate, is devoid of any transition metal ion or other cofactor and thus represents a novel type of ring-cleavage dioxygenase. Gene qdo was cloned and sequenced. Its overexpression in Escherichia coli yielded recombinant His-tagged Qdo which was catalytically active. Qdo exhibited 36% and 16% amino acid identity to 1H-3-hydroxy-4-oxoquinaldine 2,4-dioxygenase (Hod) and atropinesterase (a serine hydrolase), respectively. Qdo as well as Hod possesses a SXSHG motif, resembling the motif GXSXG of the serine hydrolases which comprises the active-site nucleophile (X=arbitrary residue).
Max
N
N
Universität Hohenheim, Institut für Mikrobiologie, D-70593, Stuttgart, Germany.
Betz
A
A
Facey
S
S
Lingens
F
F
Hauer
B
B
Fetzner
S
S
D000595:Amino Acid Sequence; D001483:Base Sequence; D049308:Dioxygenases; D008969:Molecular Sequence Data; D010105:Oxygenases / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D016958:Pseudomonas putida / Q000201:enzymology / Q000235:genetics*; D016415:Sequence Alignment
D010105:Oxygenases; C101931:1H-3-hydroxy-4-oxoquinoline 2,4-dioxygenase (decyclizing, CO-forming); D049308:Dioxygenases