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Author(s)
Giovanni Bertoni,
Fabrizio Bolognese,
E. Galli,
Paola Barbieri
Publication year1996
Publication title
applied and environmental microbiology
Resource typeJournals
PublisherAmerican Society for Microbiology
In order to study the toluene and o-xylene catabolic genes of Pseudomonas stutzeri OX1, a genomic library was constructed. A 28-kb EcoRI restriction endonuclease DNA fragment, cloned into the vector plasmid pLAFR1 and designated pFB3401, permitted Pseudomonas putida PaW340 to convert toluene and o-xylene into the corresponding meta-ring fission products. Physical and functional endonuclease restriction maps have been derived from the cloned DNA fragment. Further subcloning into and deletion analysis in the Escherichia coli vector pGEM-3Z allowed the genes for the conversion of toluene or o-xylene into the corresponding catechols to be mapped within a 6-kb region of the pFB3401 insert and their direction of transcription to be determined. Following exposure to toluene, E. coli cells carrying this 6-kb region produce a mixture of o-cresol, m-cresol, and p-cresol, which are further converted to 3-methylcatechol and 4-methylcatechol. Similarly, a mixture of 2,3-dimethylphenol and 3,4-dimethylphenol, further converted into dimethylcatechols, was detected after exposure to o-xylene. The enzyme involved in the first step of toluene and o-xylene degradation exhibited a broad substrate specificity, being able to oxidize also benzene, ethylbenzene, m-xylene, p-xylene, styrene, and naphthalene. Deletions of the 6-kb region which affect the ability to convert toluene or o-xylene into the corresponding methylphenols compromise also their further oxidation to methylcatechols. This suggests that a single enzyme system could be involved in both steps of the early stages of toluene and o-xylene catabolism.
Subject(s)bacteria , biochemistry , biology , chemistry , gene , genetics , haeiii , microbiology and biotechnology , organic chemistry , polymerase chain reaction , pseudomonas putida , pseudomonas stutzeri , restriction fragment length polymorphism , toluene , xylene
Language(s)English
SCImago Journal Rank1.552
H-Index324
eISSN1070-6291
pISSN0099-2240
DOI10.1128/aem.62.10.3704-3711.1996
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