Novel aerobic 2-aminobenzoate metabolism. Purification and characterization of 2-aminobenzoate-CoA ligase, localisation of the gene on a 8-kbp plasmid, and cloning and sequencing of the gene from a denitrifying Pseudomonas sp.
A new pathway for the aerobic metabolism of 2-aminobenzoate which proceeds via 2-aminobenzoyl-CoA has recently been revealed in a Pseudomonas strain KB 740-. The enzyme catalyzing the first step, the formation of the coenzyme A (CoA) thioester of 2-aminobenzoate, is 2-aminobenzoate-CoA ligase. It was purified from cells aerobically grown with 2-aminobenzoate as sole carbon, energy, and nitrogen source and characterized. It is rather specific for 2-aminobenzoate, but activates also benzoate and fluorobenzoates. ATP was cleaved into AMP and pyrophosphate. The ligase is a monomer of M(r) 65,000, as determined by gel filtration and SDS/PAGE. The N-terminal amino acid sequence was determined and the gene locus of the enzyme was identified by Southern blot hybridization on a small 8-kbp plasmid pKB 740. The 1.8-kb nucleotide sequence of the 2-aminobenzoate-CoA ligase gene and the derived amino acid sequence of the native enzyme (597 residues) are reported.
Altenschmidt
U
U
Abteilung Angewandte Mikrobiologie, Universität Ulm, Federal Republic of Germany.
Fuchs
G
G
D000332:Aerobiosis; D000595:Amino Acid Sequence; D001483:Base Sequence; D002845:Chromatography; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003001:Cloning, Molecular; D003066:Coenzyme A Ligases / Q000235:genetics* / Q000302:isolation & purification* / Q000378:metabolism; D004269:DNA, Bacterial / Q000235:genetics; D017886:Durapatite; D005798:Genes, Bacterial; D006882:Hydroxyapatites; D007700:Kinetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010957:Plasmids; D011549:Pseudomonas / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development; D012689:Sequence Homology, Nucleic Acid; D062367:ortho-Aminobenzoates / Q000378:metabolism*
D004269:DNA, Bacterial; D006882:Hydroxyapatites; D062367:ortho-Aminobenzoates; C031385:anthranilic acid; D017886:Durapatite; C070685:2-aminobenzoate coenzyme A ligase; D003066:Coenzyme A Ligases