Svetoch E.A., Stern N.J., Eruslanov B.V., Kovalev Y.N., Volodina L.I., Perelygin V.V.,
Mitsevich E.V., Mitsevich I.P., Pokhilenko V.D., Borzenkov V.N., Levchuk V.P., Svetoch O.E., Kudriavtseva T.Y. |
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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. |
