title: Role of lipase from community-associated methicillin-resistant Staphylococcus aureus strain USA300 in hydrolyzing triglycerides into growth-inhibitory free fatty acids.
abstract: Part of the human host innate immune response involves the secretion of bactericidal lipids on the skin and delivery of triglycerides into abscesses to control invading pathogens. Two Staphylococcus aureus lipases, named SAL1 and SAL2, were identified in the community-associated methicillin-resistant S. aureus strain USA300, which, presumably, are produced and function to degrade triglycerides to release free fatty acids. We show that the SAL2 lipase is one of the most abundant proteins secreted by USA300 and is proteolytically processed from the 72-kDa proSAL2 to the 44-kDa mature SAL2 by the metalloprotease aureolysin. We show that spent culture supernatants had lipase activity on both short- and long-chain fatty acid substrates and that deletion of gehB, encoding SAL2, resulted in the complete loss of these activities. With the use of gas chromatography-mass spectrometry, we show that SAL2 hydrolyzed trilinolein to linoleic acid, a fatty acid with known antistaphylococcal properties. When added to cultures of USA300, trilinolein and, to a lesser extent, triolein inhibited growth in a SAL2-dependent manner. This effect was shown to be due to the enzymatic activity of SAL2 on these triglycerides, since the catalytically inactive SAL2 Ser412Ala mutant was incapable of hydrolyzing the triglycerides or yielding delayed growth in their presence. Overall, these results reveal that SAL2 hydrolyzes triglycerides of both short- and long-chain fatty acids and that the released free fatty acids have the potential to cause significant delays in growth, depending on the chemical nature of the free fatty acid.
authors.affiliation: Department of Microbiology & Immunology, University of Western Ontario, London, Ontario, Canada.
authors.forename: Brigitte
authors.initials: B
authors.lastname: Cadieux
authors.affiliation: Department of Microbiology & Immunology, University of Western Ontario, London, Ontario, Canada.
authors.forename: Vithooshan
authors.initials: V
authors.lastname: Vijayakumaran
authors.affiliation: Department of Biology, University of Western Ontario, London, Ontario, Canada.
authors.forename: Mark A
authors.initials: MA
authors.lastname: Bernards
authors.affiliation: Department of Microbiology & Immunology, University of Western Ontario, London, Ontario, Canada Centre for Human Immunology, University of Western Ontario, London, Ontario, Canada.
authors.forename: Martin J
authors.initials: MJ
authors.lastname: McGavin
authors.affiliation: Department of Microbiology & Immunology, University of Western Ontario, London, Ontario, Canada Centre for Human Immunology, University of Western Ontario, London, Ontario, Canada deh@uwo.ca.
authors.forename: David E
authors.initials: DE
authors.lastname: Heinrichs
mesh_terms: D000900:Anti-Bacterial Agents / Q000378:metabolism*; D017714:Community-Acquired Infections / Q000382:microbiology; D005230:Fatty Acids, Nonesterified / Q000378:metabolism*; D006801:Humans; D006868:Hydrolysis; D008049:Lipase / Q000737:chemistry / Q000378:metabolism*; D055624:Methicillin-Resistant Staphylococcus aureus / Q000201:enzymology* / Q000302:isolation & purification; D008970:Molecular Weight; D011499:Protein Processing, Post-Translational; D014280:Triglycerides / Q000378:metabolism*
chemical_list: D000900:Anti-Bacterial Agents; D005230:Fatty Acids, Nonesterified; D014280:Triglycerides; D008049:Lipase