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Applied and Environmental Microbiology, May 1999, p. 2128-2135, Vol. 65, No. 5
Centre for Food and Animal Research,
Agriculture and Agri-Food Canada, Ottawa, Ontario K1A 0C6, Canada
Received 14 October 1998/Accepted 26 February 1999
The ruminal anaerobe Butyrivibrio fibrisolvens OR79
produces a bacteriocin-like activity demonstrating a very broad
spectrum of activity. An inhibitor was isolated from spent culture
fluid by a combination of ammonium sulfate and acidic
precipitations, reverse-phase chromatography, and high-resolution gel
filtration. N-terminal analysis of the isolated inhibitor yielded a
15-amino-acid sequence (G-N/Q-G/P-V-I-L-X-I-X-H-E-X-S-M-N). Two
different amino acid residues were detected in the second and third
positions from the N terminus, indicating the presence of
two distinct peptides. A gene with significant homology to one
combination of the determined N-terminal sequence was cloned, and
expression of the gene was confirmed by Northern blotting. The
gene (bvi79A) encoded a prepeptide of 47 amino acids and a
mature peptide, butyrivibriocin OR79A, of 25 amino acids.
Significant sequence homology was found between this peptide and
previously reported lantibiotics containing the double-glycine leader
peptidase processing site. Immediately downstream of bvi79A
was a second, partial open reading frame encoding a peptide with
significant homology to proteins which are believed to be involved in
the synthesis of lanthionine residues. These findings indicate that the
isolated inhibitory peptides represent new lantibiotics.
Results from both total and N-terminal amino acid sequencing indicated
that the second peptide was identical to butyrivibriocin OR79A except
for amino acid substitutions in positions 2 and 3 of the mature
lantibiotic. Only a single coding region was detected when restriction
enzyme digests of total DNA were probed either with an oligonucleotide
based on the 5' region of bvi79A or with degenerate
oligonucleotides based on the predicted sequence of the second peptide.
0099-2240/99/$04.00+0
Evidence for Production of a New Lantibiotic (Butyrivibriocin
OR79A) by the Ruminal Anaerobe Butyrivibrio fibrisolvens
OR79: Characterization of the Structural Gene Encoding
Butyrivibriocin OR79A

*
Corresponding author. Present address: Lethbridge
Research Centre, P. O. Box 3000, 5403 1st Ave. S., Lethbridge,
Alberta T1J 4B1, Canada. Phone: (403) 317-2246. Fax: (403) 382-3156. E-mail: teather{at}em.agr.ca.
Lethbridge Research Centre contribution no. 3879911.
Present address: Microbiology Research Division, Health Protection
Branch, Health Canada, Tunney's Pasture, Ottawa, Ontario K1A 0L2, Canada.
§
Present address: Lethbridge Research Centre, Agriculture and
Agri-Food Canada, Lethbridge, Alberta T1J 4B1, Canada.
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