Five independent clones of the P-37 lipase gene () were obtained in and the gene was localized to a 2.75 kb fragment by subcloning. The five clones were shown to contain the same gene by Southern blotting with a DIG-labelled probe to The nucleotide sequence of was determined, and shown to contain a single ORF of 1017 kb, encoding a protein of 339 amino acids. The predicted molecular mass was 36 kDa. A 33 kDa (PAGE) radiolabeled polypeptide was detected from minicell preparations harbouring which could correspond to GehA after cleavage of the putative 26 amino acid residue signal peptide. was overexpressed in under the control of the bacteriophage T7 promoter, and the corresponding polypeptide was found to be present in insoluble aggregates. Active lipase was produced when the overexpressing strain was incubated at a reduced temperature in the presence of sucrose. Purification of lipase from culture supernatant fluids confirmed the production of a 33 kDa (PAGE) lipase.


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