JCBPS; Special Issue, Section A ; 30 Nov. 2014 Vol. 4, No. 5, 136.

E- ISSN: 2249 1929

Journal of Chemical, Biological and Physical Sciences

An International Peer Review E-3 Journal of Sciences
Available online atwww.jcbsc.org

Section A: Food Biotechnology

CODEN (USA): JCBPAT

Research abstract

Partial Characterization of Antilisterial Activity Produced

by two Lactic-Acid Bacterium Strains
J.C. Ramrez-Palacios, J. Zapata-Lara, M.R. Lpez-Cuellar, A.I. Rodrguez-Hernndez,
N. Chavarra-Hernndez
1

Cuerpo Acadmico de Biotecnologa Agroalimentaria, Instituto de Ciencias

Agropecuarias, Universidad Autnoma del Estado de Hidalgo. Tulancingo, Hidalgo, CP 43600,
Abstract: Bacteriocins are ribosomally-sinthesized peptides mainly from lactic-acid bacteria
(LAB), showing antimicrobial activity against related microorganisms. Most LAB bacteriocins
are non-toxic to eukaryotic cells and are generally recognized as safe (GRAS). These peptides
have attracted considerable interest given their potential to be used in food preservation and health
care applications. The aim of this study was to identify and partially purify bacteriocin type
substances (PPB) produced by Enterococcus faecium and Streptococcus infantarius. Both strains
were cultured in MRS medium at 35C for 24 h. The PPB were obtained by centrifugation (cellfree supernatant) and adjusted pH to 6.5. The cell-free supernatant was concentrated and
pasteurized (110C, 10 min). A two-fold dilution series of PPB were prepared to check the activity
against Listeria monocytogenes as indicator, using a commercial bacteriocin (Nisin) as a
reference. The antilisterial activity were 13,900 and 3,200 AU/mL for S. infantarius and E.
faecium, respectively (also equivalent to 44,200 and 437 mg nisin/mL, respectively). Nowadays
there are no reports about bacteriocins produced by S. infantarius. Further studies must be done
to fully characterize these bacteriocins.
Keywords: bacteriocin, Listeria monocytogenes, antimicrobial activity, antilisterial

Corresponding author: Chavarra-Hernndez N

Corresponding
author: N. Chavarra-Hernndez
*norberto@uaeh.edu.mx

136

JCBPS; Section A: Food Biotechnogy; Special Issue; 30 Nov. 2014, Vol. 4, No. 5, 136

Tepetitla de Lardizbal, Tlaxcala, August 15, 2014.

ACEPTANCE LETTER
Dear colleagues:

We are pleased to inform that your work entitled:

PARTIAL CHARACTERIZATION OF ANTILISTERIAL ACTIVITY PRODUCED BY TWO

LACTIC ACID BACTERIUM STRAINS.
J.C. Ramrez-Palacios, J. Zapata-Lara, M.R. Lpez-Cuellar, A.I. Rodrguez-Hernndez, N.
Chavarra-Hernndez.

Has been accepted by the Scientific Committee, for presenting at the 1st Biotechnology
World Symposium and 9 Encuentro Nacional de Biotecnologa del IPN, which will take
place on October 13-16, 2014 at Tlaxcala, Mxico. After August 30th you can consult date,
time and presentation form of your work at the web site:
http://www.9encuentro.cibatlaxcala.ipn/ProgramaGeneral.aspx

Thank you for your invaluable participation.

PARTIAL CHARACTERIZATION OF ANTILISTERIAL ACTIVITY PRODUCED BY TWO LACTIC

ACID BACTERIUM STRAINS
J.C. Ramrez-Palacios, J. Zapata-Lara, M.R. L pez-Cuellar, A.I. Rodrguez-Hernndez, N. Chavarra-Hernndez
Cuerpo Acadmico de Biotecnologa Agroalimentaria, Instituto de Ciencias Agropecuarias, Universidad Autnoma del
Estado de Hidalgo. Tulancingo, Hidalgo, CP 43600, norberto@uaeh.edu.mx
Key words: bacteriocin, Listeria, antimicrobial activity
Antilisterial activity of bacteriocin from S. infantarius has
Introduction. Bacteriocins are ribosomally-synthesized
not been reported before. In this work, it exhibited a
peptides mainly from lactic acid bacteria (LAB), showing
greater antimicrobial activity (13,900 AU/mL) than that
observed for E. faecium (3,200 AU/mL) (Table 1).
antimicrobial activity against related microorganisms (1).
Bacteriocins have attracted considerable interest given
Chakchouk-Mtibaa (1) reported an enterocin (FL31) with a
their potential to be used in food preservation and health
lower antilisterial activity (290 AU/mL).
care. Bacteriocins are specific-target molecules with a
narrow-spectrum than that of antibiotics (2).
Streptococcus infantarius, isolated from Mexican pozol is
a LAB that produces a heat-stable peptide with high
antimicrobial activity against Listeria monocytogenes, a
human pathogen.
Enterococci are LAB, generally recognized as safe
(GRAS), found in animal intestinal tracts as well as food
products and are known to produce a wide range of
enterocins (3).
Fig. 2 Semilogarithmic plot of activity units (AU) - diameter (mm) of
In this study, we have partially purified and characterized
inhibition of PBB against L. monocytogenes.
bacteriocins from S. infantarius and Enterococcus
faecium, both showing antilisterial activity.
Table 1. Antilisterial activity of the partially purified bacteriocins.
Methods. S. infantarius (donated by G. Daz, FQ-UNAM)
and E. faecium were cultured during 24 h in MRS broth at
30 and 37 C, respectively. Mediums were adjusted to pH
6.5 and centrifuged (6,000 rpm, 4C, 15 min). Supernatant
were concentrated, pasteurized (110C, 10 min) and
stored at -80C until use. Activity against L.
monocytogenes was evaluated in BHI-agar spot assay.
These partially purified bacteriocins (PPB) were tested
after treatment with proteinase K (60 C, 1 h), -amilase
(85C, 1 h) and peptidase (37C, 1h). Arbitrary activity
units (AU) were calculated (4). Nisin was used as control.
Results.

S. infantarius
PPB
PPB + proteinase K
PPB + -amilase
PPB + peptidase
E. faecium
PPB
PPB + proteinase
PPB + -amilase
PPB + peptidase

Diameter (mm)
20.01
NA
16.63
17.51
Diameter (mm)
14.46
NA
13.8
13.16

AU/mL
13,900
3,700
5,300
AU/mL
3,200
2,500
2,000

mg nisin/mL
44,240
5,314
11,046
mg nisin/mL
437
505
296

NA: No antimicrobial activity detected

Conclusions. Both antimicrobials produced showed

antilisterial activity. After treatment with proteinase K, no
antimicrobial activity was detected, as expected due to
their peptidic nature. Further studies must be done to fully
characterize these bacteriocins.
Acknowledgements.

Veranos Cientficos AMC and Delfn 2014;

PROMEP Red Temtica 2011; Posdoctorado PROMEP 2013.

References.
Fig.1 a) Two-fold dilution method showing antilisterial activity for
bacteriocin produced by a) S. infantarius and b) E. faecium

We considered an activity unit as the reciprocal of the

highest dilution showing a clear inhibition zone (Fig. 1 and
2).

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Mellouli, L (2014). Anaerobe. 27: 1-6
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