A comparison of methods used for testing staphylokinase (fibrinolysin) production in Staphylococcus strains

Devriese, Luc; Kerckhove, A. Van De

doi:10.1007/bf00395826

Cited by 20 publications

(11 citation statements)

References 11 publications

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“…The purity of the recombinant protein was analyzed on 15% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) [23], and the concentration of the purified SRH was estimated by Lowry's method [24] with bovine serum albumin (BSA) as a standard. The activity of un-induced and induced protein under different induced concentrations was tested on heated plasma agar plate as described in previous study [25]. …”

Section: Methodsmentioning

confidence: 99%

In VitroCharacterization of a Multifunctional Staphylokinase Variant with Reduced Reocclusion, Produced from Salt InducibleE. coliGJ1158

Pulicherla

Kumar

Gadupudi

et al. 2013

BioMed Research International

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The thrombolytic therapy with clinically approved drugs often ensues with recurrent thrombosis caused by thrombin-induced platelet aggregation from the clot debris. In order to minimize these problems, a staphylokinase (SAK)-based bacterial friendly multifunctional recombinant protein SRH (staphylokinase (SAK) linked with tripeptide RGD and dodecapeptide Hirulog (SRH)) was constructed to have Hirulog as an antithrombin agent and RGD (Arg-Gly-Asp) as an antiplatelet agent in the present study. This multifunctional fusion protein SRH was expressed in osmotically inducible E. coli GJ1158 as soluble form and purified with a yield of 0.27 g/L and functionally characterized in vitro. SRH retained the fibrinolytic activity and plasminogen activation rate comparable to the parental counterpart SAK. The antithrombin activity of SRH was significantly higher than SAK. The platelet rich clot lysis assay indicated that SRH had enhanced platelet binding activity and T 50% and C50 of SRH were significantly lower than that of SAK. Furthermore, SRH inhibited the ADP-induced platelet aggregation in dose-dependent manner while SAK had no significant effect on platelet aggregation. Thus, the current study suggests that the SAK variant produced from osmotically inducible GJ1158 is more potent thrombolytic agent with antithrombin and antiplatelet aggregation activities for reduction of reocclusion in thrombolytic therapy.

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Section: Methodsmentioning

confidence: 99%

In VitroCharacterization of a Multifunctional Staphylokinase Variant with Reduced Reocclusion, Produced from Salt InducibleE. coliGJ1158

Pulicherla

Kumar

Gadupudi

et al. 2013

BioMed Research International

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“…aureus strains and their lysogenized and cured derivatives was tested on fibrin agar plates, with and without added canine serum 0.5% (v/v) as plasminogen source, as described by Devriese & Van de Kerckhove (1980). Staphylokinase converts plasminogen to its active form plasmin, which can degrade fibrin.…”

Section: Introductionmentioning

confidence: 99%

Staphylococcus aureus Bacteriophages Mediating the Simultaneous Lysogenic Conversion of -Lysin, Staphylokinase and Enterotoxin A: Molecular Mechanism of Triple Conversion

et al. 1989

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A new group of serotype F bacteriophages of Staphylococcus aureus has been found which mediates the simultaneous triple-lysogenic conversion of enterotoxin A, staphylokinase and beta-lysin. The phages were recovered fro methicillin-resistant strains of S. aureus isolated in Irish hospitals between 1971 and 1988 and from strain PS42-D, which has been used as the propagating strain for the S. aureus typing phage 42D since before 1965. The molecular mechanism of triple conversion mediated by three of these phages was determined by molecular cloning, restriction endonuclease site mapping and hybridization analysis, and compared with the mechanism of beta-lysin and staphylokinase conversion mediated by the serotype F, double-converting phase phi 13. THe genetic determinants mediating expression of enterotoxin A (entA) and staphylokinase (sak) were cloned from the DNA of the triple-converting phage and expression of the cloned determinants detected in Escherichia coli and S. aureus. The entA and sak determinants were closely linked in the phage DNA adjacent to the phage attachment site (attP) in each case and furthermore, the sak determinant of phage phi 13 was also located near its attP. The restriction maps of the entA-, sak- and attP-containing DNA regions of the three triple-converting phages were very similar to each other and to the corresponding sak- and attP- containing DNA region of phage phi 13. Hybridization analysis using a cloned beta-lysin determinant (hlb) and cloned attP-containing DNA fragments as probes demonstrated that beta-lysin conversion mediated by the triple-converting phages and phage phi 13 was caused by insertional inactivation of the chromosomally encoded hlb determinant by orientation-specific integration of phage DNA following lysogenization.

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“…saprophyticus and Staph. epidermidis were also genetically heterogeneous and many new species were described associated with man (Kloos & Schleifer 1975aSchleifer & Kloos 1975), with animals (Hajek 1976;Kloos et al 1976a;Devriese et al 1978Devriese et al , 1983 and with meat products (Schleifer & Fischer 1982). Strains identified as Peptococcus saccharolyticus were shown to be anaerobic staphylococci and they may constitute a new species (Kilpper-Balz & Schleifer 198 1); strains catalogued as micrococci belonged to the separate species Staphylococcus caseolyticus .…”

mentioning

confidence: 99%

“…sciuri and Staph. lentus (Schleifer et al 1983). Useful reviews of the classification of staphylococci are given by Kloos (1980) and Hill (1981).…”

mentioning

confidence: 99%

Identification of coagulase‐negative staphylococci from farm animals

Devriese

Schleifer

Adegoke

1985

Journal of Applied Bacteriology

126

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The species identify of 661 strains of coagulase-negative staphylococci isolated from the skin and nares of cattle, pigs, poultry, goats and sheep was determined. They belonged either to the novobiocin-sensitive species Staphylococcus hyicus, Staph. simulans, Staph. epidermidis, Staph. haemolyticus and Staph. warneri or to the novobiocin-resistant species Staph. sciuri, Staph. lentus, Staph. xylosus, Staph. cohnii, Staph. saprophyticus and Staph. gallinarum; twenty-one strains remained unidentified. The staphylococcal flora of the farm animals studied differed markedly from that associated with man; several species which do not occur in man were isolated and novobiocin-resistant strains, which occur infrequently in man, were present in large numbers in animals. Two simplified schemes for the identification of staphylococci from farm animals and man are presented.

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A comparison of methods used for testing staphylokinase (fibrinolysin) production in Staphylococcus strains

Cited by 20 publications

References 11 publications

In VitroCharacterization of a Multifunctional Staphylokinase Variant with Reduced Reocclusion, Produced from Salt InducibleE. coliGJ1158

In VitroCharacterization of a Multifunctional Staphylokinase Variant with Reduced Reocclusion, Produced from Salt InducibleE. coliGJ1158

Staphylococcus aureus Bacteriophages Mediating the Simultaneous Lysogenic Conversion of -Lysin, Staphylokinase and Enterotoxin A: Molecular Mechanism of Triple Conversion

Identification of coagulase‐negative staphylococci from farm animals

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