| http://purl.uniprot.org/citations/8407791 | http://www.w3.org/1999/02/22-rdf-syntax-ns#type | http://purl.uniprot.org/core/Journal_Citation |
| http://purl.uniprot.org/citations/8407791 | http://www.w3.org/1999/02/22-rdf-syntax-ns#type | http://purl.uniprot.org/core/Journal_Citation |
| http://purl.uniprot.org/citations/8407791 | http://www.w3.org/2000/01/rdf-schema#comment | "The genes that encode the alpha and beta subunits of protocatechuate 3,4-dioxygenase (3,4-PCD [EC 1.13.11.3]) were cloned from a Pseudomonas putida (formerly P. aeruginosa) (ATCC 23975) genomic library prepared in lambda phage. Plaques were screened by hybridization with degenerate oligonucleotides designed using known amino acid sequences. A 1.5-kb SmaI fragment from a 15-kb primary clone was subcloned, sequenced, and shown to contain two successive open reading frames, designated pcaH and pcaG, corresponding to the beta and alpha subunits, respectively, of 3,4-PCD. The amino acid sequences deduced from pcaHG matched the chemically determined sequence of 3,4-PCD in all except three positions. Cloning of pcaHG into broad-host-range expression vector pKMY319 allowed high levels of expression in P. putida strains, as well as in Proteus mirabilis after specific induction of the plasmid-encoded nahG promoter with salicylate. The recombinant enzyme was purified and crystallized from P. mirabilis, which lacks an endogenous 3,4-PCD. The physical, spectroscopic, and kinetic properties of the recombinant enzyme were indistinguishable from those of the wild-type enzyme. Moreover, the same transient enzyme intermediates were formed during the catalytic cycle. These studies establish the methodology which will allow mechanistic investigations to be pursued through site-directed mutagenesis of P. putida 3,4-PCD, the only aromatic ring-cleaving dioxygenase for which the three-dimensional structure is known."xsd:string |
| http://purl.uniprot.org/citations/8407791 | http://purl.org/dc/terms/identifier | "doi:10.1128/jb.175.19.6194-6202.1993"xsd:string |
| http://purl.uniprot.org/citations/8407791 | http://purl.org/dc/terms/identifier | "doi:10.1128/jb.175.19.6194-6202.1993"xsd:string |
| http://purl.uniprot.org/citations/8407791 | http://purl.uniprot.org/core/author | "Laporte D.C."xsd:string |
| http://purl.uniprot.org/citations/8407791 | http://purl.uniprot.org/core/author | "Laporte D.C."xsd:string |
| http://purl.uniprot.org/citations/8407791 | http://purl.uniprot.org/core/author | "Livingston D.M."xsd:string |
| http://purl.uniprot.org/citations/8407791 | http://purl.uniprot.org/core/author | "Livingston D.M."xsd:string |
| http://purl.uniprot.org/citations/8407791 | http://purl.uniprot.org/core/author | "Lipscomb J.D."xsd:string |
| http://purl.uniprot.org/citations/8407791 | http://purl.uniprot.org/core/author | "Lipscomb J.D."xsd:string |
| http://purl.uniprot.org/citations/8407791 | http://purl.uniprot.org/core/author | "Frazee R.W."xsd:string |
| http://purl.uniprot.org/citations/8407791 | http://purl.uniprot.org/core/author | "Frazee R.W."xsd:string |
| http://purl.uniprot.org/citations/8407791 | http://purl.uniprot.org/core/date | "1993"xsd:gYear |
| http://purl.uniprot.org/citations/8407791 | http://purl.uniprot.org/core/date | "1993"xsd:gYear |
| http://purl.uniprot.org/citations/8407791 | http://purl.uniprot.org/core/name | "J. Bacteriol."xsd:string |
| http://purl.uniprot.org/citations/8407791 | http://purl.uniprot.org/core/name | "J. Bacteriol."xsd:string |
| http://purl.uniprot.org/citations/8407791 | http://purl.uniprot.org/core/pages | "6194-6202"xsd:string |
| http://purl.uniprot.org/citations/8407791 | http://purl.uniprot.org/core/pages | "6194-6202"xsd:string |
| http://purl.uniprot.org/citations/8407791 | http://purl.uniprot.org/core/title | "Cloning, sequencing, and expression of the Pseudomonas putida protocatechuate 3,4-dioxygenase genes."xsd:string |
| http://purl.uniprot.org/citations/8407791 | http://purl.uniprot.org/core/title | "Cloning, sequencing, and expression of the Pseudomonas putida protocatechuate 3,4-dioxygenase genes."xsd:string |
| http://purl.uniprot.org/citations/8407791 | http://purl.uniprot.org/core/volume | "175"xsd:string |
| http://purl.uniprot.org/citations/8407791 | http://purl.uniprot.org/core/volume | "175"xsd:string |
| http://purl.uniprot.org/citations/8407791 | http://www.w3.org/2004/02/skos/core#exactMatch | http://purl.uniprot.org/pubmed/8407791 |
| http://purl.uniprot.org/citations/8407791 | http://www.w3.org/2004/02/skos/core#exactMatch | http://purl.uniprot.org/pubmed/8407791 |