Staphylococcal acid phosphatase: extensive purification and characterization of the loosely bound enzyme.
Acid phosphatase of Staphylococcus aureus PS55 was eluted from the surface of these cells with 1.0 m KCl at pH 8.5 by gentle agitation at 25 C and was purified 44-fold (51% recovery) by two cycles of dialysis and gel filtration. The eluted enzyme which had a 280/260 (nm) absorbancy ratio of 0.71 required at least 0.5 m salt solution for solubilization; however, most of the purified product which had a 280/260 (nm) absorbancy ratio of 1.72 was soluble in dilute buffer solution [0.01 m tris(hydroxymethyl)aminomethane chloride, pH 8.5]. Purified acid phosphatase appeared homogeneous according to the criteria of gel filtration, starch-block electrophoresis, and analytical ultracentrifugation. In a starch block, migration was toward the cathode at pH 8.0. Maximal activity occurred at pH 5.2 to 5.3 and salt concentration had little effect on phosphatase activity up to 1.0 m KCl or NaCl. Progressive loss of enzymatic acitivity occurred at higher salt concentrations. Molecular weight of purified acid phosphatase was estimated to be 58,000.
D000135:Acid Phosphatase / Q000032:analysis / Q000302:isolation & purification*; D002850:Chromatography, Gel; D003956:Dialysis; D004586:Electrophoresis; D004798:Enzymes / Q000032:analysis; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D013053:Spectrophotometry; D013210:Staphylococcus / Q000201:enzymology*; D014461:Ultracentrifugation
D004798:Enzymes; D000135:Acid Phosphatase