Purification and some properties of the staphylococcal extracellular lipase.

Staphylococcal lipase has been purified by application of a multistep procedure involving ammonium sulfate precipitation, and hydrophobic interaction chromatography followed by gel filtration through Sepharose CL-4B. A purified enzyme was obtained which appeared to be homogeneous by molecular sieving, polyacrylamide gel electrophoresis and sucrose gradient centrifugation. The enzyme was then subjected to physicochemical analysis. It has been found that staphylococcal lipase appears in two molecular forms: light (45 kDa) and heavy (300 kDa). Amino acid analysis indicates that lipase contains 17 amino acids with a prevalence of hydrophobic amino acids. No sulfur-containing amino acid was found in the enzyme molecule. The lipase contains about 2% sugars and some amount of lipids. The lipase preparation is stable within pH 5.0 to 9.0 and exhibits maximal activity at pH 8.0. The optimal temperature for the enzymatic reaction was established at 55 degrees C.