title: Isolation of Bacillus circulans and Paenibacillus polymyxa strains inhibitory to Campylobacter jejuni and characterization of associated bacteriocins.
abstract: We evaluated anti-Campylobacter activity among 365 Bacillus and Paenibacillus isolates from poultry production environments. One novel antagonistic Bacillus circulans and three Paenibacillus polymyxa strains were identified and further studied. Cell-free ammonium sulfate precipitate (crude antimicrobial preparation) was obtained from each candidate culture. Zones of Campylobacter growth inhibition surrounding 10 microl of this crude antimicrobial preparation were quantified using a spot test. Campylobacter growth resumed when the preparation was preincubated with selected protease enzymes, demonstrating peptide characteristics consistent with a bacteriocin. These peptides were further purified using combinations of molecular mass resolution and ion exchange chromatography. Molecular masses of the peptides were estimated at approximately 3,500 Da by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Isoelectric focusing was used to determine the pI values of the peptides. Amino acid sequences of the bacteriocins and more precise molecular masses were obtained by matrix-assisted laser desorption and ionization-time of flight (MALDI-TOF) analysis. The bacteriocin from P. polymyxa NRRL B-30507 had a pI of 4.8, that from P. polymyxa NRRL B-30509 had a pI of 7.2, that from P. polymyxa NRRL B-30508 had a pI of 4.8, and that from B. circulans NRRL B-30644 had a pI of 7.8. The amino acid sequences were consistent with those of class IIa bacteriocins. These antagonists and the corresponding bacteriocins may be useful in the control of Campylobacter infection in poultry.
authors.affiliation: State Research Center for Applied Microbiology, Obolensk, Russian Federation.
authors.forename: Edward A
authors.initials: EA
authors.lastname: Svetoch
authors.forename: Norman J
authors.initials: NJ
authors.lastname: Stern
authors.forename: Boris V
authors.initials: BV
authors.lastname: Eruslanov
authors.forename: Yuri N
authors.initials: YN
authors.lastname: Kovalev
authors.forename: Larisa I
authors.initials: LI
authors.lastname: Volodina
authors.forename: Vladimir V
authors.initials: VV
authors.lastname: Perelygin
authors.forename: Evgeni V
authors.initials: EV
authors.lastname: Mitsevich
authors.forename: Irina P
authors.initials: IP
authors.lastname: Mitsevich
authors.forename: Victor D
authors.initials: VD
authors.lastname: Pokhilenko
authors.forename: Valery N
authors.initials: VN
authors.lastname: Borzenkov
authors.forename: Vladimir P
authors.initials: VP
authors.lastname: Levchuk
authors.forename: Olga E
authors.initials: OE
authors.lastname: Svetoch
authors.forename: Tamara Y
authors.initials: TY
authors.lastname: Kudriavtseva
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001407:Bacillus / Q000302:isolation & purification / Q000502:physiology*; D001426:Bacterial Proteins / Q000302:isolation & purification / Q000378:metabolism; D001430:Bacteriocins / Q000302:isolation & purification* / Q000378:metabolism; D002169:Campylobacter Infections / Q000517:prevention & control / Q000662:veterinary*; D016123:Campylobacter jejuni / Q000254:growth & development* / Q000378:metabolism; D015169:Colony Count, Microbial; D004591:Electrophoresis, Polyacrylamide Gel; D007525:Isoelectric Focusing; D008970:Molecular Weight; D011200:Poultry; D011201:Poultry Diseases / Q000517:prevention & control*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D001426:Bacterial Proteins; D001430:Bacteriocins