Biochemical and genetic evidence for production of enterocins A and B by Enterococcus faecium WHE 81

Saïd Ennahar, Yuji Asou, Takeshi Zendo, Kenji Sonomoto, Ayaaki Ishizaki

Research output: Contribution to journalArticle

94 Citations (Scopus)

Abstract

Enterococcus faecium WHE 81, isolated from cheese, has been reported to produce a bacteriocin called enterocin 81 [J. Appl. Microbiol. 85 (1998) 521.]. Purification of enterocin 81 was carried out using ammonium sulfate precipitation, desalting on ODP-90 reverse-phase column, and purification through SP Sepharose HP cation exchange and C2/C18 reverse-phase chromatographies. The antimicrobial was eluted from the C2/C18 column as four individually active fractions, designated A81, B81, C81 and D81. The purification procedure used proved particularly efficient for the bacteriocin in fraction D81, with a yield of 46%, while only 3.8% the bacteriocin in fraction B81 could be collected. MALDI-TOF mass spectrometry of the bacteriocins in fractions B81 and D81 showed respective masses of 4833.0 and 5462.2 Da. Amino acid sequencing of the two peptides revealed two class-II bacteriocins whose sequences were similar to those of enterocin A and enterocin B, respectively. Using proper primers, chromosomal fragments of 212 and 216 bp enclosing bacteriocin structural genes were PCR-amplified. Cloning of the amplicons and their sequencing revealed two genes with sequences identical to the structural genes of enterocins A and B, respectively. It was therefore clearly established that E. faecium WHE 81 produces bacteriocins respectively identical to enterocins A and B. Our results, combined with data from previous reports, suggest that the two bacteriocins may be widespread among enterococcal strains and may play an important role in controlling the growth of pathogens and other undesirable bacteria in certain fermented food products.

Original languageEnglish
Pages (from-to)291-301
Number of pages11
JournalInternational Journal of Food Microbiology
Volume70
Issue number3
DOIs
Publication statusPublished - Nov 8 2001

Fingerprint

enterocins
Enterococcus faecium
Bacteriocins
bacteriocins
Molecular Biology
structural genes
Genes
fermented foods
enterocin A
enterocin B
matrix-assisted laser desorption-ionization mass spectrometry
reversed-phase liquid chromatography
cation exchange
Matrix-Assisted Laser Desorption-Ionization Mass Spectrometry
Protein Sequence Analysis
Ammonium Sulfate
Cheese
Reverse-Phase Chromatography
ammonium sulfate
Sepharose

All Science Journal Classification (ASJC) codes

  • Food Science
  • Microbiology

Cite this

Biochemical and genetic evidence for production of enterocins A and B by Enterococcus faecium WHE 81. / Ennahar, Saïd; Asou, Yuji; Zendo, Takeshi; Sonomoto, Kenji; Ishizaki, Ayaaki.

In: International Journal of Food Microbiology, Vol. 70, No. 3, 08.11.2001, p. 291-301.

Research output: Contribution to journalArticle

Ennahar, Saïd ; Asou, Yuji ; Zendo, Takeshi ; Sonomoto, Kenji ; Ishizaki, Ayaaki. / Biochemical and genetic evidence for production of enterocins A and B by Enterococcus faecium WHE 81. In: International Journal of Food Microbiology. 2001 ; Vol. 70, No. 3. pp. 291-301.
@article{0825141a22f44cd58ea2164443c220c5,
title = "Biochemical and genetic evidence for production of enterocins A and B by Enterococcus faecium WHE 81",
abstract = "Enterococcus faecium WHE 81, isolated from cheese, has been reported to produce a bacteriocin called enterocin 81 [J. Appl. Microbiol. 85 (1998) 521.]. Purification of enterocin 81 was carried out using ammonium sulfate precipitation, desalting on ODP-90 reverse-phase column, and purification through SP Sepharose HP cation exchange and C2/C18 reverse-phase chromatographies. The antimicrobial was eluted from the C2/C18 column as four individually active fractions, designated A81, B81, C81 and D81. The purification procedure used proved particularly efficient for the bacteriocin in fraction D81, with a yield of 46{\%}, while only 3.8{\%} the bacteriocin in fraction B81 could be collected. MALDI-TOF mass spectrometry of the bacteriocins in fractions B81 and D81 showed respective masses of 4833.0 and 5462.2 Da. Amino acid sequencing of the two peptides revealed two class-II bacteriocins whose sequences were similar to those of enterocin A and enterocin B, respectively. Using proper primers, chromosomal fragments of 212 and 216 bp enclosing bacteriocin structural genes were PCR-amplified. Cloning of the amplicons and their sequencing revealed two genes with sequences identical to the structural genes of enterocins A and B, respectively. It was therefore clearly established that E. faecium WHE 81 produces bacteriocins respectively identical to enterocins A and B. Our results, combined with data from previous reports, suggest that the two bacteriocins may be widespread among enterococcal strains and may play an important role in controlling the growth of pathogens and other undesirable bacteria in certain fermented food products.",
author = "Sa{\"i}d Ennahar and Yuji Asou and Takeshi Zendo and Kenji Sonomoto and Ayaaki Ishizaki",
year = "2001",
month = "11",
day = "8",
doi = "10.1016/S0168-1605(01)00565-7",
language = "English",
volume = "70",
pages = "291--301",
journal = "International Journal of Food Microbiology",
issn = "0168-1605",
publisher = "Elsevier",
number = "3",

}

TY - JOUR

T1 - Biochemical and genetic evidence for production of enterocins A and B by Enterococcus faecium WHE 81

AU - Ennahar, Saïd

AU - Asou, Yuji

AU - Zendo, Takeshi

AU - Sonomoto, Kenji

AU - Ishizaki, Ayaaki

PY - 2001/11/8

Y1 - 2001/11/8

N2 - Enterococcus faecium WHE 81, isolated from cheese, has been reported to produce a bacteriocin called enterocin 81 [J. Appl. Microbiol. 85 (1998) 521.]. Purification of enterocin 81 was carried out using ammonium sulfate precipitation, desalting on ODP-90 reverse-phase column, and purification through SP Sepharose HP cation exchange and C2/C18 reverse-phase chromatographies. The antimicrobial was eluted from the C2/C18 column as four individually active fractions, designated A81, B81, C81 and D81. The purification procedure used proved particularly efficient for the bacteriocin in fraction D81, with a yield of 46%, while only 3.8% the bacteriocin in fraction B81 could be collected. MALDI-TOF mass spectrometry of the bacteriocins in fractions B81 and D81 showed respective masses of 4833.0 and 5462.2 Da. Amino acid sequencing of the two peptides revealed two class-II bacteriocins whose sequences were similar to those of enterocin A and enterocin B, respectively. Using proper primers, chromosomal fragments of 212 and 216 bp enclosing bacteriocin structural genes were PCR-amplified. Cloning of the amplicons and their sequencing revealed two genes with sequences identical to the structural genes of enterocins A and B, respectively. It was therefore clearly established that E. faecium WHE 81 produces bacteriocins respectively identical to enterocins A and B. Our results, combined with data from previous reports, suggest that the two bacteriocins may be widespread among enterococcal strains and may play an important role in controlling the growth of pathogens and other undesirable bacteria in certain fermented food products.

AB - Enterococcus faecium WHE 81, isolated from cheese, has been reported to produce a bacteriocin called enterocin 81 [J. Appl. Microbiol. 85 (1998) 521.]. Purification of enterocin 81 was carried out using ammonium sulfate precipitation, desalting on ODP-90 reverse-phase column, and purification through SP Sepharose HP cation exchange and C2/C18 reverse-phase chromatographies. The antimicrobial was eluted from the C2/C18 column as four individually active fractions, designated A81, B81, C81 and D81. The purification procedure used proved particularly efficient for the bacteriocin in fraction D81, with a yield of 46%, while only 3.8% the bacteriocin in fraction B81 could be collected. MALDI-TOF mass spectrometry of the bacteriocins in fractions B81 and D81 showed respective masses of 4833.0 and 5462.2 Da. Amino acid sequencing of the two peptides revealed two class-II bacteriocins whose sequences were similar to those of enterocin A and enterocin B, respectively. Using proper primers, chromosomal fragments of 212 and 216 bp enclosing bacteriocin structural genes were PCR-amplified. Cloning of the amplicons and their sequencing revealed two genes with sequences identical to the structural genes of enterocins A and B, respectively. It was therefore clearly established that E. faecium WHE 81 produces bacteriocins respectively identical to enterocins A and B. Our results, combined with data from previous reports, suggest that the two bacteriocins may be widespread among enterococcal strains and may play an important role in controlling the growth of pathogens and other undesirable bacteria in certain fermented food products.

UR - http://www.scopus.com/inward/record.url?scp=0035829346&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0035829346&partnerID=8YFLogxK

U2 - 10.1016/S0168-1605(01)00565-7

DO - 10.1016/S0168-1605(01)00565-7

M3 - Article

C2 - 11764194

AN - SCOPUS:0035829346

VL - 70

SP - 291

EP - 301

JO - International Journal of Food Microbiology

JF - International Journal of Food Microbiology

SN - 0168-1605

IS - 3

ER -