Isolation and purification of staphylococcal lipase.
An extracellular lipase was isolated from the cell-free supernatant fluid of a 24-hr culture of Staphylococcus aureus grown in Trypticase Soy Broth at 37 C with continuous agitation. The purification was achieved by precipitation with alcohol followed by differential precipitation at pH 8.6 and 4.3. Subsequent purification with Sephadex G 200 and BioGel 300 yielded a preparation which showed a 350- to 450-fold increase in specific activity over the original cell-free supernatant fluid. The purified lipase was homogeneous over a BioGel 300 column and showed a single peak on electrophoresis in a Veronal buffer (pH 8.6, Gamma/2 = 0.1). The electrophoretic mobility was -7.78 x 10(-5) cm(2) per v per sec.
D002498:Centrifugation; D002845:Chromatography; D008049:Lipase / Q000032:analysis*; D013210:Staphylococcus / Q000201:enzymology*; D014461:Ultracentrifugation
D008049:Lipase