title: The Pseudomonas aeruginosa devB/SOL homolog, pgl, is a member of the hex regulon and encodes 6-phosphogluconolactonase.
abstract: A cyclic version of the Entner-Doudoroff pathway is used by Pseudomonas aeruginosa to metabolize carbohydrates. Genes encoding the enzymes that catabolize intracellular glucose to pyruvate and glyceraldehyde 3-phosphate are coordinately regulated, clustered at 39 min on the chromosome, and collectively form the hex regulon. Within the hex cluster is an open reading frame (ORF) with homology to the devB/SOL family of unidentified proteins. This ORF encodes a protein of either 243 or 238 amino acids; it overlaps the 5' end of zwf (encodes glucose-6-phosphate dehydrogenase) and is followed immediately by eda (encodes the Entner-Doudoroff aldolase). The devB/SOL homolog was inactivated in P. aeruginosa PAO1 by recombination with a suicide plasmid containing an interrupted copy of the gene, creating mutant strain PAO8029. PAO8029 grows at 9% of the wild-type rate using mannitol as the carbon source and at 50% of the wild-type rate using gluconate as the carbon source. Cell extracts of PAO8029 were specifically deficient in 6-phosphogluconolactonase (Pgl) activity. The cloned devB/SOL homolog complemented PAO8029 to restore normal growth on mannitol and gluconate and restored Pgl activity. Hence, we have identified this gene as pgl and propose that the devB/SOL family members encode 6-phosphogluconolactonases. Interestingly, three eukaryotic glucose-6-phosphate dehydrogenase (G6PDH) isozymes, from human, rabbit, and Plasmodium falciparum, contain Pgl domains, suggesting that the sequential reactions of G6PDH and Pgl are incorporated in a single protein. 6-Phosphogluconolactonase activity is induced in P. aeruginosa PAO1 by growth on mannitol and repressed by growth on succinate, and it is expressed constitutively in P. aeruginosa PAO8026 (hexR). Taken together, these results establish that Pgl is an essential enzyme of the cyclic Entner-Doudoroff pathway encoded by pgl, a structural gene of the hex regulon.
authors.affiliation: Department of Microbiology and Immunology, The Brody School of Medicine, East Carolina University, Greenville, NC 27858-4354, USA.
authors.forename: P W
authors.initials: PW
authors.lastname: Hager
authors.forename: M W
authors.initials: MW
authors.lastname: Calfee
authors.forename: P V
authors.initials: PV
authors.lastname: Phibbs
mesh_terms: D000446:Aldehyde-Lyases / Q000235:genetics*; D000595:Amino Acid Sequence; D001483:Base Sequence; D002265:Carboxylic Ester Hydrolases / Q000235:genetics*; D017124:Conserved Sequence; D005634:Fructose-Bisphosphate Aldolase; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D005942:Gluconates / Q000378:metabolism; D005954:Glucosephosphate Dehydrogenase / Q000235:genetics; D008353:Mannitol / Q000378:metabolism; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D011550:Pseudomonas aeruginosa / Q000201:enzymology / Q000235:genetics*; D018085:Regulon; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid
chemical_list: D005942:Gluconates; D008353:Mannitol; D005954:Glucosephosphate Dehydrogenase; D002265:Carboxylic Ester Hydrolases; C022363:6-phosphogluconolactonase; D000446:Aldehyde-Lyases; D005634:Fructose-Bisphosphate Aldolase; C100131:phospho-2-keto-3-deoxy-gluconate aldolase; C030691:gluconic acid