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PMID: 8953719 Published · ppublish English Journal Article

Analysis of cumene (isopropylbenzene) degradation genes from Pseudomonas fluorescens IP01.

Applied and environmental microbiology ·Vol. 62 ·No. 12 ·1996-12-00 ·Pages 4471-7

Habe H, Kasuga K, Nojiri H, Yamane H, Omori T

Abstract

We obtained the DNA fragments encoding 2-hydroxy-6-oxo-7-methylocta-2,4-dienoic acid (HOMODA) hydrolase in the cumene (isopropylbenzene) degrader Pseudomonas fluorescens strain IP01 via PCR using two synthesized oligonucleotides corresponding to the conserved regions within known meta-cleavage compound hydrolases. Following colony hybridization using the amplified DNA as a probe, a 4.5-kb HindIII fragment was isolated from P. fluorescens IP01. After determining the nucleotide sequence of this fragment, three open reading frames (ORF11 [cumH], ORF12 [cumD], and ORF13) were identified. The deduced amino acid sequence of ORF12 showed homology with meta-cleavage compound hydrolases encoded by the tod, dmp, xyl, and bph operons. Although the product of ORF12 was found to exhibit HOMODA and 2-hydroxy-6-oxohepta-2,4-dienoic acid (HOHDA) hydrolase activities, it did not exhibit 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid (HOPDA) hydrolase activity. The deduced amino acid sequence of ORF11 showed 40.4% homology with the sequence of todX in Pseudomonas putida F1 (Y. Wang, M. Ralings, D. T. Gibson, D. Labbé, H. Bergeron, R. Brousseau, and P. C. K. Lau, Mol. Gen. Genet. 246:570-579, 1995). The nucleotide sequence of ORF13 and its flanking region showed strong homology (91.0%) with IS52 from Pseudomonas savastanoi (Y. Yamada, P.-D. Lee, and T. Kosuge, Proc. Natl. Acad. Sci. USA 83:8263-8267, 1982). By characterization of cumH and cumD, the entire cum gene cluster from the cumene-degrader P. fluorescens IP01 (cumA1A2A3A4BCEGFHD) has been identified.

MeSH Terms
Amino Acid Sequence Base Sequence Benzene Derivatives/metabolism Cloning, Molecular Genes, Bacterial Molecular Sequence Data Open Reading Frames Pseudomonas/genetics
Chemicals
Benzene Derivatives cumene
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Habe H
Biotechnology Research Center, University of Tokyo, Japan.
Kasuga K
Nojiri H
Yamane H
Omori T
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31 references, click to expand
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Article Info
Journal
Applied and environmental microbiology
Abbr.
Appl Environ Microbiol
ISSN
0099-2240
DOI
10.1128/aem.62.12.4471-4477.1996
Published
1996-12-00
Pages
4471-7
Language
English
Region
United States
NLM ID
7605801
PMCID
PMC168274
Subset
IM
Databases
GENBANK
D83955
External Links
PubMed source DOI: 10.1128/aem.62.12.4471-4477.1996 PMC Full Text View journal details
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