Purification and properties of boticin P produced by Clostridium botulinum

Abstract

A bacteriocin produced spontaneously by a nontoxigenic strain of Clostridium botulinum, type E, PM-15, has been isolated and designated boticin P. It was purified by ammonium sulfate precipitation, gel filtration on Sephadex G-100, sucrose density gradient centrifugation, and cesium chloride equilibrium density gradient centrifugation. Boticin P is composed mainly of proteins (98.8%) with a trace amount of carbohydrates (0.4%), and has an apparent molecular weight in excess of 4 × 106 daltons as estimated by gel filtration on Sepharose 2B. Electron microscopic examination of boticin P reveals a phage tail-like structure of 100 nm in length.Boticin P exerted a static effect on vegetative growth and spore outgrowth but not on the initial events of germination. The boticin was active on 10/12 toxigenic and 3/6 nontoxigenic type E and 2/2 nonproteolytic type B strains of C. botulinum. The activity spectrum on 27 strains supports the proposal that type E and the nonproteolytic type B strains belong to the same taxosubspecies.

Butyricin 7423: a Bacteriocin Produced by Clostridium butyricum NCIB7423

“…Production of bacteriocins by species of Clostridium is not uncommon: boticins are formed by strains of Clostridium botulinum (Ellison & Kautter, 1970;Lau, Hawirko & Chow, 1974) ; perfringocins by strains of Clostridiurn perfringens (Tubylewicz, I 966 ;; and other clostridocins are secreted by several unidentified nonpathogenic Clostridium species (Hongo et al, 19684 b). Yet until recently, only the mode of action of perfringocin 28 had been studied in any detail, its bacteriolytic action being attributed either to inhibition of cell-wall synthesis or to removal of the existing wall (Mahony, Butler & Lewis, 1971;Mahony, 1973).…”

Clostridium botulinumandClostridium tetani

“…The four groups also have distinctive surface antigen relationships (Batty and Walker 1966;Hatheway 1983, 1984a, b;Lynt et al 1967;Solomon et al 1969Solomon et al , 1971Walker and Batty 1964) and by the morphology and host range of bacteriophages and antibacterial spectra of bacteriocins (boticins) (Dineen et al 2000;Eklund et al 1988;Graham 1978;Hall and Peterson 1922;Kautter et al 1966;Kiritani et al 1973;Lau et al 1974;Mahony 1979;Smith 1975).…”

“…Bacteriocins and other uncharacterized antimicrobial substances are produced by a number of clostridia including C. botulinum (Dineen et al 2000;Eklund et al 1988;Graham 1978;Hall and Peterson 1922;Kautter et al 1966;Kiritani et al 1973;Lau et al 1974;Mahony 1979;Smith 1975). Smith (1975) isolated strains of B. cereus, C. sporogenes, and C. perfringens that produced bacteriocins active against C. botulinum.…”

“…Another inhibitory factor was less heat resistant and was reported to have a molecular weight of 4 Â 10 7 Da. A bacteriocin-like substance was isolated from a nontoxigenic type E-like strain which was proposed to resemble a defective phage (Lau et al 1974). Bacteriophages have been isolated from non-proteolytic C. botulinum (Sugiyama and King 1972).…”

Purification of Two Clostridium Bacteriocins by Procedures Appropriate to Hydrophobic Proteins

“…Bacteriocins effective against obligately anaerobic organisms have been particularly neglected. Yet, several species of Clostridium are known to produce bacteriocins (2,6,14,15,21). The most studied of these are the perfringocins which are produced by strains of Clostridium perfringens and are active on many other strains of this species (25)(26)(27)36).…”