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Applied and Environmental Microbiology, December 1998, p. 4883-4890, Vol. 64, No. 12
Departamento de Biotecnología de
Alimentos, Instituto de la Grasa, Consejo Superior de
Investigaciones Científicas, 41012 Seville, Spain
Received 12 March 1998/Accepted 11 August 1998
Enterocin I (ENTI) is a novel bacteriocin produced by
Enterococcus faecium 6T1a, a strain originally isolated
from a Spanish-style green olive fermentation. The bacteriocin is
active against many olive spoilage and food-borne
gram-positive pathogenic bacteria, including clostridia,
propionibacteria, and Listeria monocytogenes. ENTI was
purified to homogeneity by ammonium sulfate precipitation, binding to
an SP-Sepharose fast-flow column, and phenyl-Sepharose CL-4B and
C2/C18 reverse-phase chromatography. The
purification procedure resulted in a final yield of 954% and a
170,000-fold increase in specific activity. The primary structure of
ENTI was determined by amino acid and nucleotide sequencing. ENTI
consists of 44 amino acids and does not show significant sequence
similarity with any other previously described bacteriocin. Sequencing
of the entI structural gene, which is located on the 23-kb
plasmid pEF1 of E. faecium 6T1a, revealed the absence of a
leader peptide at the N-terminal region of the gene product. A second
open reading frame, ORF2, located downstream of entI,
encodes a putative protein that is 72.7% identical to ENTI.
entI and ORF2 appear to be cotranscribed, yielding an mRNA
of ca. 0.35 kb. A gene encoding immunity to ENTI was not identified.
However, curing experiments demonstrated that both enterocin production
and immunity are conferred by pEF1.
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Purification and Genetic Characterization of
Enterocin I from Enterococcus faecium 6T1a, a Novel
Antilisterial Plasmid-Encoded Bacteriocin Which Does Not Belong
to the Pediocin Family of Bacteriocins
*
Corresponding author. Mailing address: Departamento de
Biotecnología de Alimentos, Instituto de la Grasa, Consejo
Superior de Investigaciones Cientificas, Aptdo. 1078, 41012 Seville,
Spain. Phone: 34-5-4692516. Fax: 34-5-4691262. E-mail address:
rjimenez{at}cica.es.
Applied and Environmental Microbiology, December 1998, p. 4883-4890, Vol. 64, No. 12
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
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