Production, purification, and preliminary characterization of a gonococcal growth inhibitor produced by a coagulase-negative staphylococcus isolated from the urogenital flora

Abstract

Coagulase-negative staphylococcus no. 7, isolated from the urogenital flora, was grown on semisolid brain heart infusion medium. Supernatants were obtained by centrifuging frozen and thawed media which had supported the growth of the staphylococci at 37 degrees C. The kinetic of production revealed that the antigonococcal activity was detected at the end of the logarithmic phase of growth and that the maximum activity was obtained after 24 h of incubation. Production of inhibitory activity was detected in cultures grown between 35 and 39 degrees C and in a pH range of 6.9 to 9.4. The inhibitory substance was purified by methanol extraction, acetone fractionation, dialysis, and chromatography on Ultrogel AcA 54. The characterization of the inhibitor showed that it was a lipoprotein or a lipid-associated-protein and that the protein component could be separated from the lipids when chromatographed on Ultrogel AcA 54 in the presence of urea. The inhibitory activity was associated with the protein component which had a molecular weight of approximately 15,900. In polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate and urea, the protein component was dissociated in a subunit estimated to be approximately 1,400 daltons.

Relative Quantitative Comparisons of the Extracellular Protein Profiles ofStaphylococcus aureusUAMS-1 and ItssarA,agr, andsarA agrRegulatory Mutants Using One-Dimensional Polyacrylamide Gel Electrophoresis and Nanocapillary Liquid Chromatography Coupled with Tandem Mass Spectrometry

“…3b). The amino acid sequence of this peptide was found to be similar (54 to 76% amino acid identity) to three highly related peptides (63) originally identified in Staphylococcus haemolyticus as a "gonococcal growth inhibitor" (1). Since that time, these peptides have been found in a number of coagulase-negative staphylococci that exhibit a synergistic zone of complete hemolysis with betahemolysin-producing strains on sheep blood, much like the delta-hemolysin of S. aureus (15,16,29).…”

Antimicrobial Activity of Community-associated Methicillin-resistant Staphylococcus aureus Is Caused by Phenol-soluble Modulin Derivatives

“…In addition, it has recently been shown that specific S. epidermidis PSMs, S. epidermidis PSM␦ and ␦-toxin (PSM␥), have antimicrobial activities against Streptococcus pyogenes owing to their capacity to form pores in the bacterial cell membrane (22). Finally, antimicrobial activities have been detected sporadically for other staphylococcal PSM-like peptides (23). Whether S. aureus PSMs are antimicrobial and responsible for antimicrobial activities exhibited by some S. aureus strains is unknown.…”

The amino acid sequence of a gonococcal growth inhibitor from Staphylococcus haemolyticus

“…The production and purification of the gonococcal inhibitor were carried out as previously described (Beaudet et al, 1982 Elution was with an acetonitrile gradient (1 %/min) at 1.5 ml/min from a Synchropak RP-P column equilibrated in 0.05 % trifluoroacetic acid. The letter T denotes trypsin cleavage.…”

“…7 was used for the production of the bactericidal inhibitor (Bisaillon et al, 1980;Lafond et al, 1981;Beaudet et al, 1982). The reference target strain was Neisseria gonorrhoeae G-10 and it was cultivated as previously described (Frenette et al, 1984).…”

“…The purification of a bactericidal inhibitor produced by coagulase-negative staphylococci no. 7 was described by Beaudet et al (1982). The characterization of the purified substance revealed that it was a lipoprotein or a lipid-associated protein in which the protein component is the active part.…”