ARS | Publication request: ISOLATION OF BACILLUS CIRCULANS AND PAENIBACILLUS POLYMYXA INHIBITORY TO CAMPYLOBACTER JEJUNI AND CHARACTERIZATION OF ASSOCIATED BACTERIOCINS

Submitted to: Campylobacter Helicobacter and Related Organisms International Workshop
Publication Type: Abstract Only
Publication Acceptance Date: July 1, 2003
Publication Date: September 10, 2003
Citation: Stern, N.J., Svetoch, E.A., Eruslanov, B.V., Kovalev, Y.N., Yolodina, L.I., Perelygin, V.V., Mitsevich, E.V., Mitsevich, I.P., Pokhilenko, V.D., Borzenkov, V.N. 2003. Isolation of bacillus circulans and paenibacillus polymyxa inhibitory to campylobacter jejuni and characterization of associated bacteriocins. Campylobacter Helicobacter and Related Organisms International Workshop.

Technical Abstract: We evaluated anti-Campylobacter activity among 365 Bacillus and Paenibacillus strains isolated from poultry production environments. We measured zones of inhibition surrounding the candidate strains on agar lawns of Campylobacter. One novel antagonistic Bacillus circulans (NRRL B-30644) and two Paenibacillus polymyxa (NRRL B-30507 & NRRL B-30509) strains were identified as important and deposited under provisions of the Budapest Treaty. The cell-free, ammonium sulfate precipitate from each candidate culture was termed the crude antimicrobial preparation (CAP). Zones of Campylobacter growth inhibition surrounding 10 ul of the CAP were observed. Campylobacter growth resumed when the CAP was pre-incubated with protease enzymes, thus demonstrating a peptide characteristic consistent with bacteriocins definition. These peptides were further purified using a combination of ammonium sulfate precipitation, CM-Sepharose, Superose, and ion exchange chromatography. Molecular weights of the peptides were determined by SDS-PAGE electrophoresis. Isoelectric focusing was used to determine the isoelectric points (pI) of the peptides. Amino acid sequences of the bacteriocins were determined. The bacteriocin from strain B-30507 had an estimated molecular weight of 3.5 kDa and a pI of 4.8; the bacteriocins from strain B-30509 had corresponding values of 3.5 kDa and a pI of 7.2; and the bacteriocin from strain B-30644 had corresponding values of 3.5 kDa and a pI of 7.8. These antagonists and the corresponding bacteriocin products may be useful in the control of Campylobacter in poultry.