Cloning and nucleotide sequences of the genes involved in the meta-cleavage pathway of cumene degradation in Pseudomonas fluorescens IP01

A SalI fragment which contained the downstream region of a gene cluster encoding enzymes degrading cumene (isopropylbenzene) to 2-hydroxy-6-oxo-7-methylocta-2,4-dienoic acid was cloned from Pseudomonas fluorescens IP01. We determined the nucleotide sequence of the 4,045-bp region, and identified three open reading frames (ORFs). Comparison of amino acid sequences encoded by the three ORFs with those of enzymes encoded by the bph, dmp, xyl and tod operons suggested that the ORFs encode enzymes which are required for conversion of 2-hydroxypenta-2,4-dienoic acid to Krebs cycle intermediates in the meta-cleavage pathway. Taken together with the previously characterized cumA1A2(ORF3)A3A4BC genes, ten genes involved in cumene degradation have been identified.