Occurrence of a Complex Form of Staphylokinase in the Course of Cultivation of Staphylococcus aureus

Abstract

Proteins in the culture supernatant of Staphylococcus aureus PS 47 were subjected to Sephadex chromatography. In the early stage of the cultivation, staphylokinase appeared to have a molecular weight of 15,000, and in the later stage it appeared to have a molecular weight of 320,000. The staphylokinase having a lower molecular weight (type A) converted into one having a higher value (type B) during the course of cultivation. It was demonstrated that conversion of type A into type B took place in vitro (monitored by gel filtration after the two types of staphylokinases were mixed), and it was observed that type B reverted to type A when type B was treated with KCl or detergent. Type B seems to be a complex form of type A and some high-molecular-weight substance.

Purification, Characterization, and Evaluation of Fibrinolytic Activity of Staphylokinase From Locally Isolated Staphylococcus Aureus Gh38

“…In addition, the inhibition of the enzyme by PMSF suggests that the purified enzyme was a serine protease. Also, reduction by 2-Mercaptoethanol is one of the most common agents used for disulfide reduction because cleavage of a single disulfide bond Fujimura et al (12). Present results concord many of the earlier studies on the fibrinolytic enzyme.…”

Purification and partial characterization of a lysine-specific protease ofPorphyromonas gingivalis

“…The activity against other p-nitroanilide derivatives of amino acids or peptides was deter-134 mined by the same methods as described above. Fibrinolytic activity was examined according to the method of assay of plasmin activity [17] with modifications. The reaction mixture consisted of 100 /xl of enzyme (0.61 Tosyl-Gly-Pro-Lys-pNA hydrolytic activity ml-~), 200 pA of 0.4% bovine fibrinogen (plasminogen-free, Daiichi Pure Chemical, Tokyo, Japan), 200 Izl of 50 U m1-1 bovine thrombin (Mochida Pharmaceutical Co., Tokyo, Japan), and 500 /xl of 50 mM Tris-HCl buffer (pH 8.0).…”

Cloning, high-level expression, purification and characterization of a staphylokinase variant, SakC, from Staphylococcus aureus QT08 in Escherichia coli BL21

“…Meanwhile, after incubation for 8 h, the rSak activity retained above 50% in comparison to the original activity ( Figure 4A), but less than 20% at temperature range of 60 to 70°C. rSak from S. aureus QT08 showed a similar profile in the temperature stability to native Sak (Fujimura et al, 1974;Schlott et al, 1994b). The activity of the native Sak from S. aureus after heating at 60°C for 2 h decreased to 11% of that of the original sample (Fujimura et al, 1974).…”

“…rSak from S. aureus QT08 showed a similar profile in the temperature stability to native Sak (Fujimura et al, 1974;Schlott et al, 1994b). The activity of the native Sak from S. aureus after heating at 60°C for 2 h decreased to 11% of that of the original sample (Fujimura et al, 1974). Both wild type Sak42D and SakSAR showed a different thermal stability; Sak42D was inactivated more rapidly while SakSAR gradually inactivated for several hours at high temperature (70°C) (Schlott et al, 1994b).…”