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PMID: 9758777 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't

Analysis of the gene cluster encoding toluene/o-xylene monooxygenase from Pseudomonas stutzeri OX1.

Applied and environmental microbiology ·Vol. 64 ·No. 10 ·1998-10-00 ·Pages 3626-32

Bertoni G, Martino M, Galli E, Barbieri P

Abstract

The toluene/o-xylene monooxygenase cloned from Pseudomonas stutzeri OX1 displays a very broad range of substrates and a very peculiar regioselectivity, because it is able to hydroxylate more than one position on the aromatic ring of several hydrocarbons and phenols. The nucleotide sequence of the gene cluster coding for this enzymatic system has been determined. The sequence analysis revealed the presence of six open reading frames (ORFs) homologous to other genes clustered in operons coding for multicomponent monooxygenases found in benzene- and toluene-degradative pathways cloned from Pseudomonas strains. Significant similarities were also found with multicomponent monooxygenase systems for phenol, methane, alkene, and dimethyl sulfide cloned from different bacterial strains. The knockout of each ORF and complementation with the wild-type allele indicated that all six ORFs are essential for the full activity of the toluene/o-xylene monooxygenase in Escherichia coli. This analysis also shows that despite its activity on both hydrocarbons and phenols, toluene/ o-xylene monooxygenase belongs to a toluene multicomponent monooxygenase subfamily rather than to the monooxygenases active on phenols.

MeSH Terms
Cloning, Molecular Escherichia coli Genes, Bacterial Multigene Family Open Reading Frames Oxygenases/chemistry,genetics,metabolism Plasmids Protein Biosynthesis Pseudomonas/enzymology,genetics Recombinant Proteins/metabolism Restriction Mapping Sequence Homology, Amino Acid Species Specificity Substrate Specificity
Chemicals
Recombinant Proteins Oxygenases toluene 2-xylene monooxygenase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Bertoni G
Dipartimento di Genetica e di Biologia dei Microrganismi, Università degli Studi di Milano, 20133 Milan, Italy.
Martino M
Galli E
Barbieri P
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Article Info
Journal
Applied and environmental microbiology
Abbr.
Appl Environ Microbiol
ISSN
0099-2240
DOI
10.1128/AEM.64.10.3626-3632.1998
Published
1998-10-00
Pages
3626-32
Language
English
Region
United States
NLM ID
7605801
PMCID
PMC106479
Subset
IM
Databases
GENBANK
AJ005663
External Links
PubMed source DOI: 10.1128/AEM.64.10.3626-3632.1998 PMC Full Text View journal details
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