Purification of Staphylococcal β-Hemolysin and Its Action on Staphylococcal and Streptococcal Cell Walls

Chesbro, William; Heydrick, Fred P.; Martineau, Roland; Perkins, Gail N.

doi:10.1128/jb.89.2.378-389.1965

Cited by 35 publications

(14 citation statements)

References 31 publications

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“…Proc., p. 78, 1970) isolated delta lysins with three different isoelectric points. Our findings support the idea that staphylococcal delta hemolysin is molecularly heterogeneous, i.e., it may, as staphylococcal alpha lysin (8,64) and beta lysin (13,30,48), exist in more than one active physicochemical form. Many other bacterial toxins are known to be physicochemically heterogeneous (2).…”

Section: Discussionsupporting

confidence: 87%

Purification and Properties of Staphylococcal Delta Hemolysin

et al. 1971

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Large amounts (200 mg per liter of culture supernatant fluid) of highly purified staphylococcal soluble delta hemolysin were obtained by adsorption to and selective elution from hydroxyapatite followed by exhaustive dialysis against water, concentration by polyvinylpyrrolidone or polyethylene glycol 20,000 dialysis, and a final water dialysis. No carbohydrate, phosphorus, or inactive 280-nm absorbing material was detected in the preparation; however, analysis by density gradient centrifugation, gel filtration, analytical ultracentrifugation, carboxymethyl cellulose chromatography, polyacrylamide disc gel electrophoresis, isoelectric focusing, and electron microscopy revealed that the lysin was molecularly heterogeneous. The preparation contained an acidic fibrous lysin (S20, of 11.9) and a basic lysin component composed of a population of granular aggregates of various sizes, with a maximum Sm,, of approximately 4.9. No other staphylococcal products were detected in the preparation. The lysin was active against erythrocytes from many animal species and acted synergistically with staphylococcal beta hemolysin against sheep erythrocytes. It was soluble in chloroform-methanol (2: 1), was inactivated by various phospholipids, normal sera, and proteolytic enzymes, but was partially resistant to heat inactivation. Activity was not affected by Ca2+, Mg2+, citrate, ethylenediaminetetraacetic acid, or cysteine. The lysin preparation also disrupted bacterial protoplasts and spheroplasts, erythrocyte membranes, lysosomes, and lipid spherules, was growth-inhibitory for certain bacteria, and clarified egg yolk-agar. Large amounts produced dermonecrosis in rabbits and guinea pigs. The minimum lethal intravenous dose for mice and guinea pigs was approximately 110 and 30 mg/kg, respectively. (vii) clear egg yolk-agar (41); and (viii) possess phospholipase C activity with phosphatidylinositol and phosphatidylserine as substrates (67). Rigorous proof for the belief that all of these effects are caused by delta hemolysin is lacking. Delta hemolysin has been partially purified and characterized by Marks and Vaughan (51),

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Section: Discussionsupporting

confidence: 87%

Purification and Properties of Staphylococcal Delta Hemolysin

et al. 1971

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“…ribonuclease (RNase), and deoxyribonuclease (DNase) activities were determined by methods previously used in this laboratory (6)(7)(8). All other staphylococcal products were assayed according to the methods described by Chesbro et al (9).…”

Section: Association Of Bactericidal Substance With Phage Type 71 S mentioning

confidence: 99%

Bactericidal Substance From Staphylococcus Aureus

Dajani

Gray

Wannamaker

1970

The Journal of Experimental Medicine

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The majority of Staphylococcus aureus strains recovered in pure form from superficial skin lesions have been shown to belong to phage type 71 (1, 2). Certain unique features of this phage type have been described previously, among which are the common resistance to penicillin G (1, 2), inhibition of Corynebacterium diphtheriae on solid media (1, 3), and the production of opacity in horse serum agar, but not in egg yolk broth medium (3).In a previous communication from our laboratory (4), a bactericidal substance has been described in supernatant fluids from broth cultures of phage type 71 S. aureus. This substance was active against streptococci belonging to Groups A, C, and D, pneumococci, and corynebacteria. No inhibition of Group B streptococci or Gram-negative rods was noted, and inhibition of Group G streptococci and staphylococci was variable. The bactericidal substance has been shown to be a protein or a polypeptide. The present investigations further describe the biological properties of this bactericidal substance and indicate that it is distinct from other known staphylococcal products. Materials and MethodsMedla.--Trypdc soy broth (TSB) 1 (Difco) was used to cultivate all the staphylococcal strains. A dialysate medium of TSB was prepared as previously described (4)

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“…This reduced volume was applied to a 25-cm column of cellulose phosphate (Schleicher & Schuell Co., Keene, N. H.) prepared as described elsewhere (7). The column was eluted by use of a NaCl gradient produced by a nine-chambered device (16).…”

mentioning

confidence: 99%

Enzymatic Detection of the Growth of Staphylococcus aureus in Foods1

Chesbro¹,

Auborn²

1967

Applied Microbiology

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A specific method has been developed for the extraction and measurement of staphylococcal nuclease in foods in which Staphylococcus aureus has grown. The method was used to. compare staphylococcal growth with nuclease production in foods under varying conditions of temperature, aerobiosis, and competition from other microorganisms. It was concluded that the nuclease is produced under any conditions that permit growth of S. aureus, and little or no interference with the test was encountered either from mixed, natural populations or from a variety of pure, laboratory cultures. Nuclease and enterotoxin A production were shown to vary in synchrony for the 234 (Casman) strain of S. aureus, and the sensitivity of the enzymatic detection of nuclease was comparable to the sensitivity of serological detection of enterotoxin A. It was found that 15 min at 121 C was required to reduce the nuclease activity in slurries of contaminated ham below the level present in the unheated slurry. The extraordinary heat resistance of the nuclease permits its detection even in foods heated subsequent to the growth of S. aureus. The nuclease analysis requires about 3 hr to complete and requires no unusual equipment or reagents.

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Purification of Staphylococcal β-Hemolysin and Its Action on Staphylococcal and Streptococcal Cell Walls

Cited by 35 publications

References 31 publications

Purification and Properties of Staphylococcal Delta Hemolysin

Purification and Properties of Staphylococcal Delta Hemolysin

Bactericidal Substance From Staphylococcus Aureus

Enzymatic Detection of the Growth of Staphylococcus aureus in Foods1

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