Isolation of Bacillus circulans and Paenibacillus polymyxa strains inhibitory to Campylobacter jejuni and characterization of associated bacteriocins.
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.
Svetoch
Edward A
EA
State Research Center for Applied Microbiology, Obolensk, Russian Federation.
Stern
Norman J
NJ
Eruslanov
Boris V
BV
Kovalev
Yuri N
YN
Volodina
Larisa I
LI
Perelygin
Vladimir V
VV
Mitsevich
Evgeni V
EV
Mitsevich
Irina P
IP
Pokhilenko
Victor D
VD
Borzenkov
Valery N
VN
Levchuk
Vladimir P
VP
Svetoch
Olga E
OE
Kudriavtseva
Tamara Y
TY
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
D001426:Bacterial Proteins; D001430:Bacteriocins