We report the complete nucleotide sequence and over-expression of the genes for the acetaldehyde dehydrogenase (acylating) and the 4-hydroxy-2-oxovalerate aldolase from the pathway operon of the naphthalene catabolic plasmid pWW60-22 from sp. NCIMB9816. Additional partial sequence analysis of adjacent DNA shows the gene order within the operon to be , identical to the order found for the isofunctional genes in the pathway operons in the toluene/xylene pathway of TOL plasmid pWWO and the phenol/methylphenol pathway of pVI150. The deduced amino acid sequences of NahO and NahM were significantly homologous to the equivalent enzymes encoded by other pathways, although both were the most divergent in each comparison. The genes were inserted downstream of the T7 promoter in the expression vector pET3a and similar constructs were also made of the isofunctional regions from pVI150 () and TOL plasmid pDK1 (). High expression of all three gene pairs was detected by enzyme assays and by SDS-PAGE.


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