Genetic Organization and Sequence of the 'Pseudomonas cepacia' for the Alpha and Beta Subunits of Protocatechuate 3,4-Dioxygenase

摘要

Locations of genes for alpha and beta subunits of protocatechuate 3,4-dioxygenase on a 9.5 kilobase pair PstI fragment cloned from the Pseudomonas cepacia DB01 chromosome were determined after construction of several subclones into the broad-host-range cloning vectors pR02317, pR02320, and pR02321. The ability of each subclone to complement mutations in protocatechuate 3,4-dioxygenase (pcaA) was tested in mutant strains derived from P. cepacia, P. aeruginosa, and P. putida. These complementation studies also showed that the two subunits were expressed from the same promoter. The nucleotide sequence of the region encoding for protocatechuate 3,4-dioxygenase was determined. The deduced amino acid sequence matched that determined by N-terminal analysis of regions of the isolated enzyme. The deduced amino acid sequence shows extensive homology with the previously determined amino acid sequence of protocatechuate 3,4-dioxygenase from another Pseudomonas species. (Copyright (c) 1989 American Society for Microbiology.)