Molecular analysis of a plasmid-encoded phenol hydroxylase from Pseudomonas CF600.
Pseudomonas strain CF600 is able to utilize phenol and 3,4-dimethylphenol as sole carbon and energy source. We demonstrate that growth on these substrates is by virtue of plasmid-encoded phenol hydroxylase and a meta-cleavage pathway. Screening of a genomic bank, with DNA from the previously cloned catechol 2,3-dioxygenase gene of the TOL plasmid pWW0, was used in the identification of a clone which could complement a phenol-hydroxylase-deficient transposon insertion mutant. Deletion mapping and polypeptide production analysis identified a 1.2 kb region of DNA encoding a 39.5 kDa polypeptide which mediated this complementation. Enzyme activities and growth properties of Pseudomonas strains harbouring this fragment on a broad-host-range expression vector indicate that phenol hydroxylase is a multicomponent enzyme containing the 39.5 kDa polypeptide as one component.
Shingler
V
V
Unit for Applied Cell and Molecular Biology, University of Umeå, Sweden.
Franklin
F C
FC
Tsuda
M
M
Holroyd
D
D
Bagdasarian
M
M
D001426:Bacterial Proteins / Q000235:genetics*; D015342:DNA Probes; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000235:genetics; D005798:Genes, Bacterial; D015698:Genomic Library; D006899:Mixed Function Oxygenases / Q000235:genetics*; D010455:Peptides / Q000235:genetics; D010957:Plasmids; D011549:Pseudomonas / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development
D001426:Bacterial Proteins; D015342:DNA Probes; D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D010455:Peptides; D006899:Mixed Function Oxygenases; C022336:phenol 2-monooxygenase