title: Identification of a second lipase gene, gehD, in Staphylococcus epidermidis: comparison of sequence with those of other staphylococcal lipases.
abstract: The identification and molecular characterization of a previously unidentified lipase, gehD, from the human cutaneous commensal Staphylococcus epidermidis is reported. A lipase-GehC-deficient but otherwise isogenic mutant of S. epidermidis 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, gehD, 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 gehC and gehD genes were found to be present in S. epidermidis 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 S. epidermidis may be more closely related to those of Staphylococcus aureus than to each other.
authors.affiliation: Division of Microbiology, School of Biochemistry and Molecular Biology, University of Leeds, Leeds LS2 9JT, UK1.
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Longshaw
authors.affiliation: Division of Microbiology, School of Biochemistry and Molecular Biology, University of Leeds, Leeds LS2 9JT, UK1.
authors.forename: Angela M
authors.initials: AM
authors.lastname: Farrell
authors.affiliation: Division of Microbiology, School of Biochemistry and Molecular Biology, University of Leeds, Leeds LS2 9JT, UK1.
authors.forename: John D
authors.initials: JD
authors.lastname: Wright
authors.affiliation: Division of Microbiology, School of Biochemistry and Molecular Biology, University of Leeds, Leeds LS2 9JT, UK1.
authors.forename: Keith T
authors.initials: KT
authors.lastname: Holland
mesh_terms: D000595:Amino Acid Sequence; D005798:Genes, Bacterial; D006801:Humans; D008049:Lipase / Q000235:genetics*; D008969:Molecular Sequence Data; D009154:Mutation; D010802:Phylogeny; D017384:Sequence Deletion; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D013210:Staphylococcus / Q000201:enzymology / Q000235:genetics; D013211:Staphylococcus aureus / Q000201:enzymology / Q000235:genetics; D013212:Staphylococcus epidermidis / Q000201:enzymology* / Q000235:genetics* / Q000302:isolation & purification
chemical_list: D008049:Lipase