Fatty acid oxidation in Pseudomonas and expression and partial DNA sequence of acyl-CoA synthetase in Pseudomonas aeruginosa PAK

The fatty acid oxidation pathway is required for complete degradation of lipids after their hydrolysis to fatty acids. Observation of growth on triphenyl tetrazolium chloride (TTC) fatty acid indicator agar and measurement of ;Acyl-CoA synthetase activates fatty acids into CoA thioesters prior to their oxidation. The structural gene for acyl-CoA synthetase on plasmid pUCPAKD1 from a P. aeruginosa strain PAK genomic pUC18 library has been identified by complementation of an E. coli acyl-CoA synthetase (fadD) mutant, PN325. Complementation was determined by following growth on oleate and by expression of acyl-CoA synthetase. The insert of the pUCPAKD1 plasmid has been partially sequenced and the deduced amino acid sequence aligns with and shares a significant degree of similarity to amino acids 254 to 558 of the E. coli acyl-CoA synthetase protein.