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

Applied and Environmental Microbiology
H HabeToshio Omori

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 flankin...Continue Reading

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Citations

Jan 24, 2004·Molecular Genetics and Genomics : MGG·H OginoH Ishikawa
Dec 19, 2006·Journal of Bacteriology·Masaki ShintaniHideaki Nojiri
Dec 11, 2003·Microbiology and Molecular Biology Reviews : MMBR·Hiroshi Nikaido
Aug 18, 2004·The Journal of Biological Chemistry·Isao FujiiYutaka Ebizuka
Dec 17, 2002·The Journal of General and Applied Microbiology·Kensuke Furukawa
Aug 23, 2002·Protein Science : a Publication of the Protein Society·Shinya FushinobuTakayoshi Wakagi
Feb 8, 2006·Applied and Environmental Microbiology·Aisling MooneyAlan D W Dobson
Aug 10, 2005·Canadian Journal of Microbiology·Emma R MasterWilliam W Mohn

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