Characterization of a plasmid-specified pathway for catabolism of isopropylbenzene in Pseudomonas putida RE204

Journal Article · Wed Oct 01 04:00:00 EDT 1986 · J. Bacteriol.; (United States)

OSTI ID:7024583

Eaton, R W; Timmis, K N

A Pseudomonas putida strain designated RE204, able to utilize isopropylbenzene as the sole carbon and energy source, was isolated. Tn5 transposon mutagenesis by means of the suicide transposon donor plasmid pLG221 yielded mutant derivatives defective in isopropylbenzene metabolism. These were characterized by the identification of the products which they accumulated when grown in the presence of isopropylbenzene and by the assay of enzyme activities in cell extracts. Based on the results obtained, the following metabolic pathway is proposed: isopropylbenzene ..-->.. 2,3-dihydro-2,3-dihydroxyisopropylbenzene ..-->.. 3-isopropylcatechol ..-->.. 2-hydroxy-6-oxo-7-methylocta-2,4-dienoate ..-->.. isobutyrate + 2-oxopent-4-enoate ..-->.. amphibolic intermediates. Plasmid DNA was isolated from strain RE204 and mutant derivatives and characterized by restriction enzyme cleavage analysis. Isopropylbenzene-negative isolates carried a Tn5 insert within a 15-kilobase region of a 105-kilobase plasmid designated pRE4. DNA fragments of pRE4 carrying genes encoding isopropylbenzene catabolic enzymes were cloned in Escherichia coli with various plasmid vectors. These clones were subsequently used to generate a transposon insertion and restriction enzyme cleavage map of the isopropylbenzene metabolic region of pRE4.

• Research Organization: Universite de Geneve, Switzerland
• OSTI ID: 7024583
• Journal Information: J. Bacteriol.; (United States), Journal Name: J. Bacteriol.; (United States) Vol. 168:1; ISSN JOBAA
• Country of Publication: United States
• Language: English

Related Subjects

550400* -- Genetics
550700 -- Microbiology
59 BASIC BIOLOGICAL SCIENCES
AROMATICS
BACTERIA
BIODEGRADATION
BIOLOGICAL PATHWAYS
CELL CONSTITUENTS
CHEMICAL REACTIONS
CLONING
CUMENE
DECOMPOSITION
DNA
ENZYME ACTIVITY
GENETIC MAPPING
GROWTH
HYDROCARBONS
MAPPING
METABOLISM
MICROORGANISMS
MUTAGENESIS
NUCLEIC ACIDS
ORGANIC COMPOUNDS
PLASMIDS
PSEUDOMONAS