1887

Abstract

The identification and molecular characterization of a previously unidentified lipase, , from the human cutaneous commensal is reported. A lipase-GehC-deficient but otherwise isogenic mutant of 9 was constructed by allele replacement. However, the mutant was found to retain 50% of the wild-type lipase activity in liquid culture. Rescreening of a genomic library revealed the presence of a second lipase gene, , which was subsequently mapped and sequenced. In common with other staphylococcal lipases, GehD appeared to be translated as a 650–700 amino acid precursor which is processed post-translationally to an extracellular mature lipase of 360 amino acids with a size of approximately 45 kDa. Comparison of the amino acid sequence of GehD with those of other staphylococcal lipases revealed a high level of conservation between the mature lipase domains of different species. By hybridization studies, both and genes were found to be present in isolates from both clinical and non-clinical backgrounds, but neither hybridized to DNA isolated from other staphylococcal strains. Construction of a phylogenetic tree and calculation of amino acid sequence homologies between mature lipases, however, suggested that the lipases of may be more closely related to those of than to each other.

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2000-06-01
2020-01-29
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