[A contribution to purification of staphylokinase (author's transl)].
Optimal production of staphylokinase (SAK) was achieved in a casein hydrolysate yeast extract medium (LO3) containing 0.1 M Na-pyruvate. Addition of the pyruvate reduced significantly the production of protease and facilitated purification of SAK. After concentration of SAK from the culture supernatant by precipitation with ZnCl2 highly purified SAK was prepared successively using DEAE-Sephadex A 25 in a batch-procedure, by chromatography on CM-Sephadex C25, filtration with Ultrogel ACA 54, isoelectric focusing between pH 3.5 and 10 (fig. 2) and refocusing between pH 5-7 (table 1). For fractionation by focusing a device was constructed to record pH and optical density together with the fraction number in one diagramm (fig. 1). The purified SAK-preparation had a specific activity of 16.700 units per mg protein. It proved to be free of hemolysins, coagulase, leukocidin, lipase, nuclease, phosphatase, and protease. It was homogeneous upon analyses by diselectrophoresis, double immunodiffusion and immunelectrophoresis.
D002852:Chromatography, Ion Exchange; D007525:Isoelectric Focusing; D013211:Staphylococcus aureus / Q000201:enzymology; D013300:Streptokinase / Q000096:biosynthesis / Q000302:isolation & purification*
D013300:Streptokinase
Zur Reinigung von Staphylokinase.