The structural gene () coding for the extracellular lipase of PAO1 has been cloned on plasmid pSW118. Nucleotide sequence analysis revealed a gene of 936 bp. codes for a proenzyme of 311 amino acids including a leader sequence of 26 amino acids. The mature protein was predicted to have a of 30134, an isoelectric point of 5.6, and a consensus sequence (IGHSHGG) typical of lipases. Furthermore it is highly homologous (>60%) to other lipases from various pseudomonads. The gene failed to hybridize detectably with genomic DNA from other species except , even under relaxed stringency. Located 220 bp downstream of the gene, is an open reading frame (ORF2, ) which encodes a hydrophilic protein (283 amino acids; 33587) that shows some homology to the gene product of . In complementation tests of lipase-defective mutants, was shown to be necessary for expression of active extracellular lipase in PAO1.


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