Overview of Lactobacillus plantarum as a promising bacteriocins producer

among lactic acid bacteria

Sabrina da Silva Sabo, Michele Vitolo, José Manuel Domı́nguez González,

Ricardo Pinheiro de Souza Oliveira

PII: S0963-9969(14)00518-3
DOI: doi: 10.1016/j.foodres.2014.07.041
Reference: FRIN 5413

To appear in: Food Research International

Received date: 28 March 2014

Revised date: 14 July 2014
Accepted date: 24 July 2014

Please cite this article as: da Silva Sabo, S., Vitolo, M., González, J.M.D. & de
Souza Oliveira, R.P., Overview of Lactobacillus plantarum as a promising bacteri-
ocins producer among lactic acid bacteria, Food Research International (2014), doi:
10.1016/j.foodres.2014.07.041

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 ACCEPTED MANUSCRIPT
Overview of Lactobacillus plantarum as a promising bacteriocins

producer among lactic acid bacteria

Sabrina da Silva Saboa, Michele Vitoloa, José Manuel Domínguez Gonzálezb, Ricardo

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Pinheiro de Souza Oliveiraa*

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a
Biochemical and Pharmaceutical Technology Department, Faculty of Pharmaceutical

Sciences, São Paulo University, Av Prof Lineu Prestes, 580, Bl 16, 05508-900, São

Paulo, Brazil
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b
Department of Chemical Engineering, Faculty of Sciences, University of Vigo

(Campus Ourense), As Lagoas s/n, 32004 Ourense, Spain

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____________________

*Corresponding Author: Ricardo Pinheiro de Souza Oliveira.

Telephone: +55(11) 3091-0123

E-mail: [email protected]

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ABSTRACT

Chemical preservatives have been traditionally used during the manufacturing of

processed products. However, the continuous growing interest of consumers for fresh

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and natural products makes necessary to search for alternative compounds. In this

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context, food industries have been widely used lactic acid bacteria (LAB) as natural

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preservatives, due to their ability to produce antibacterial compounds such as

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bacteriocins. Similarly, pharmaceutical industries have improved the use of these

bacterial peptides, with antibacterial activity, trying to reduce the indiscriminate use of

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antibiotics in food products for human and animal consumption. Among LAB,
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Lactobacillus plantarum can be adapted to various niches thanks to its ability to ferment

a wide range of carbohydrates. Additionally, it can be used as starter culture in food

fermentations and as an ingredient for probiotic foods, contributing to the organoleptic

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characteristics of foods at the same time that prolongs the shelf-life and safety of these

products. It is also worthy the amount of valuable substances obtained from L.

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plantarum species isolated from different ecological niches, thus proving to be one of
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the most important and versatile species among LAB.

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Keywords: Lactic acid bacteria, Lactobacillus plantarum, bacteriocins, plantaricin,

purification of bacteriocins.

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1. Introduction

Industrialized food has experienced in recent years an increase in using chemical

additives in their formulations; however, consumers are becoming more and more

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worried about using these chemical additives in their diet. For this reason, there is a

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strong trend for seeking natural and fresh foods, free of chemical preservatives. This

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fact, associated with the growing demand for minimally processed foods, has

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encouraged the search for effective natural preservatives, among which, antibacterial

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compounds such as bacteriocins, fulfill these requirements (Castro, Palavecino,

Herman, Garro, & Campo, 2011).

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In order to control these facts, an alternative would be to use bacterial peptides

with antibacterial activity, such as bacteriocins (Parada, Caron, Medeiros & Soccol,
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2007).
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Bacteriocins are peptides or proteins synthesized within ribosomes and released

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into the extracellular medium by Gram-positive and Gram-negative microorganisms,

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although, those produced by lactic acid bacteria (LAB) have received a greater

attention, in recent years, due to their high potential for application in the food industry
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as natural conserving agents (Leroy & De Vuyst, 2004). Among LAB, the largest group

is the genus Lactobacillus, which comprises more than 150 different species (Siezen et

al., 2010). Some of those are associated with the promotion of health benefits to the host

(Bosch et al., 2011). Among them, it can be pointed out L. plantarum, an industrially

important microorganism that can be found and isolated from dairy products and

fermented foods such as sauerkraut, sourdough, sausages, cheeses, wines, olives and

pickled vegetables from environments such as cow-dung, silage and from sewage; as

well as from the human mouth, intestinal tract and stools (Parente et al., 2010; Hammer

& Vogel, 1995).

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In addition, certain L. plantarum strains have probiotic properties, which have

been used for the development of functional foods and potential oral vaccines (Parente

et al., 2010). To carry out such probiotic activities, the strains should have the ability to

produce substances such as bacteriocins, which offers advantages in colonization and

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competition in the gastrointestinal tract (Castro et al., 2011). Additionally, these

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compounds have bactericidal or bacteriostatic action on pathogenic bacteria, which

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includes important pathogens such as Listeria monocytogenes, Clostridium botulinum

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and Staphylococcus aureus (De Vuyst & Leroy, 2007). Recently, the food industry has

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shown an increasing interest in the use of bacteriocins as a replacement for chemical

preservatives, as they are effective at low concentrations and, when added to food, do
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not alter its sensory quality (Zacharof, Coss, Mandale, & Lovitt, 2013).

In this context, the purpose of this work is to provide, as a first step, a general
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overview of LAB along with the bacteriocins produced by these strains, before
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addressing a detailed study of L. plantarum species, itemizing the bacteriocins produced

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up to now, with aspect concerning to their biosynthesis, their main applications, mainly
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in the food and pharmaceutical industries, and the purification considered in order to

obtain a deeper knowledge of these peptide bacteriocins.

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2. General characteristics of LAB

The group of LAB associated with food includes 11 genera: Carnobacterium,

Enterococcus, Lactococcus, Lactobacillus (by far the most investigate genus),

Lactosphaera, Leuconostoc, Oenococcus, Pediococcus, Streptococcus, Vagococcus, and

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Weissella (Mogensen, Salminen, & O´Brien, 2003; Vries, Vaughan, Kleerebezem, &

Vos, 2006).

These microorganisms are Gram (+), non-sporulating, catalase-negative, acid-

resistant, pHoptimum for growing between 4.0 and 4.5, anaerobic aerotolerant, Toptimum for

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growing is 30oC (mesophilic) or 42oC (thermophilic) and can have different shapes, like

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rods (bacilli) and sphere (coccus) (Todorov & Franco, 2010).

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Lactobacillus plantarum is a facultative heterofermentative lactic acid

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bacterium, which ferments carbohydrates generally by the phosphoketolase pathway

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(PKP). The fermentation of pentoses (xylose, ribose) leads to the formation of pyruvate

and acetyl-P and their subsequent conversion to lactate and acetate, respectively.
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Hexoses (glucose, fructose, mannose) in these bacteria can be converted to lactate, CO2,

and ethanol (Mayo et al., 2010; Todorov & Franco, 2010; Hammes & Vogel, 1995). In
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addition, its genome encodes all enzymes required for the glycolysis and
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phosphoketolase pathways (Kleerebezem et al., 2003). L. plantarum has broad

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versatility, insofar as it can be found in many ecological niches, as well as in human and
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animal gastrointestinal tract (Siezen et al., 2010). The ability to inhabit different niches

is associated with its ability to ferment a variety of sugars (Prins et al., 2010). They have
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developed very efficient transport systems, which enable them to quickly take up the

necessary solutes (Salminen & Von Wright, 1993). Particularly, this flexible and

adaptive behavior of L. plantarum is reflected by the relatively large number of

regulatory and transport functions, including 25 complete sugars phosphotransferase

system (PTS) (Kleerebezem et al., 2003).

LAB are used in the food industry due to their ability to inhibit or reduce

contamination by spoilage and/or pathogens microorganisms through the production of

various antimicrobial compounds (Martinez et al., 2013a). The acidification of food –

mainly by lactic acid formation - is probably the primary factor in the inhibition of

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undesirable microorganisms. LAB reduces the pH to values close to 4 in these foods,

which hinders the survival of microorganisms sensitive to acid medium. Such condition

leads to the increase of the fermented product shelf life, when compared to the non-

fermented one (Leroy & De Vuyst, 2004).

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These bacteria are still used in the food industry for the development of

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organoleptic properties of fermented foods (Carminati et al., 2010; Todorov, LeBlanc,

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& Franco, 2012). This occurs due to the large number of glycolytic, lipolytic and

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proteolytic enzymes that transform some medium nutrients into compounds with

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sensory properties, which, in the end, gradually modify the structure and aroma of the

fermented food (Todorov et al., 2012). In addition, LAB are also used as probiotic
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starter cultures (Carminati et al., 2010).
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3. Bacteriocins from LAB

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LAB-producing bacteriocins (antimicrobial peptides synthesized in ribosomes)

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kill bacteria at much lower concentrations than eukaryotic antimicrobial peptides,

probably because they interact with a specific receptor present on target cells (Cotter et
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al., 2005; Drider, Fimland, Héchard, McMullen, & Prévost, 2006). Some studies

indicated that the 35d-plantaricin bacteriocin produced by L. plantarum 35d showed to

be active against Aeromonas hydrophila. Meanwhile bacteriocins ST28MS and

ST26MS, produced by L. plantarum isolated from syrup inhibited the growth of

Escherichia coli, Pseudomonas aeruginosa and Acinetobacter baumannii and other

Gram-positive microorganisms (Todorov & Dicks, 2004a; Messi, Bondi, Sabia, Battini,

& Manicardi, 2001).

The bacteriocin production often occurs during late log phase or early stationary

phase, and is generally influenced by quorum sensing mechanism or by any sign of

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stress (Martinez et al., 2013b). They differ from the majority of antibiotics due to their

molecular proteinaceous constitution, being rapidly degraded by proteases in the human

digestive tract (Parada et al., 2007).

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3.1. History of bacteriocins

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The first report of an antibacterial substance was conducted in 1925, when

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André Gratia published an article regarding the inhibitory capacity of E. coli on other

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strains of the same species. The produced compound, considered as the responsible for

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the inhibitory effect, was called "colicin", in reference to the producer microorganism

(Collins, Cotter, Hill, & Ross, 2010). In 1928, the ability of certain Lactococci strains to
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exert inhibition on other LAB strains was reported, and later, in 1947, Mattick and

Hirsh concentrated an inhibitory substance isolated from a strain of Lactococcus lactis

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subsp. lactis, termed nisin (Cotter et al., 2005). This bacteriocin was initially purified
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and marketed in 1953 in England and then, in 1969, was considered to be safe for use in
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food products by the Joint Food and Agriculture Organization / World Health
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Organization Expert Committee on Food Additives. It was also in 1953 that the term

"bacteriocin" was proposed for antimicrobial peptides produced by microorganisms

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(Reeves, 1965). In 1983, in Europe, nisin was added to the list of food additives and, in

1988, the American Food and Drug Administration (FDA) authorized its use in

processed cheeses (Collins et al., 2010). Although nisin is the only bacteriocin approved

by the FDA for using in foods, the pediocin, attained from strains of Pediococcus

acidilactici, Pediococcus parvulus and Lactobacillus plantarum WHE92, has also been

employed as preservative in industrialized foods (Enan, Essawy, Uyttendaele, &

Debevere, 1996; Wang & Wang, 2014). Indeed, there is a commercial pediocin – ALTA

2341®, produced by Quest International, Sarasota, Florida from P.acidilactici – used as

medium component for fermentation processes (Papagianni & Anastasiadou, 2009). As

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ALTA 2341® presented high inhibitory action on L. monocytogenes, the producer

applied for its approvation by FDA (Chen, Sebraneck, Dickson, & Mendonca, 2004).

As a remarkable achievement on bacteriocins was the attainment, by 1994, of a

lantibiotic called plantaricin C from L. plantarum LL441, a strain isolated from ripening

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cheese (González, Arca, Mayo, & Suárez, 1994).

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3.2. Classification of bacteriocins

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In spite of bacteriocins from lactic acid bacteria (LAB) differing in their

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spectrum of activity and in biochemical and genetic determinants, some common

characteristics allow divide them into four classes, based on the primary structure,
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molecular weight, heat stability and molecular organization (Cotter et al., 2005; Heng,

Wescombe, Burton, Jack, & Tagg, 2007):

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• Class I (lantibiotics): consists of linear (type A) and globular (type B) peptides,

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with low molecular weight (<5 kDa, with approximately 19 to 38 amino acids). They
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are post-translational modified peptides that contain unusual amino acids such as
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lanthionine and derivatives (Cintas, Casaus, Herranz, Nes, & Hernández, 2001; Drider

et al., 2006; Todorov, 2009). Nisin is the first and the most well-known lantibiotic
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(Ghrairi, Chaftar, & Hani, 2012);

• Class II (non-lantibiotics): composed of thermostable peptides (<10 kDa, with

approximately 37 to 48 amino acids), which, according to Drider et al., (2006) are

divided into three subclasses known as Class IIa (group of pediocin-like bacteriocins

active against Listeria spp.), Class IIb (bacteriocins requiring the union of two peptides

to completely exert antibacterial effect, e.g., lactocin G) and Class IIc (bacteriocins

which have a covalent bond between C and N terminal, resulting in a cyclic structure)

(Balciunas et al., 2013);

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• Class III: represented by thermolabile peptides of high molecular weight (> 30

kDa), such as helveticin J, acidophilucin A, and lactacin A and B (Heng, et al., 2007);

• Class IV: composed of complex bacteriocins that contain carbohydrate or lipid

moieties, in addition to the protein portion (Heng et al., 2007). However, Cleveland,

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Montville, Nes, and Chikindas (2001) proposed that these complexes are artifacts of

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partial purification and not a new class of bacteriocins.

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Among all of these peptide bacteriocins produced by LAB, subclass IIa has

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arisen as one of the most interesting group for use in food preservation (inhibiting the

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growth of gram-positive food spoilage and pathogenic bacteria such as Bacillus cereus,

Clostridium perfringens, S. aureus, and L. monocytogenes) as well as in medicine (as

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antibiotic complements in treating infectious diseases or as antiviral agents) (Drider et

al., 2006). Some bacteriocins produced by L. plantarum have been ascribed to this
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group, including plantaricin 423 produced by L. plantarum strain 423. This bacteriocin
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was able to inhibit the growth of L. monocytogenes in ostrich salami meat (Dicks,
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Mellett, & Hoffman, 2004).

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Other plantaricins, such as EF, JK, NC8 and J51, were cataloged into the two-

peptide bacteriocins (subclass IIb) (Anderssen, Diep, Nes, Eijsink, & Meyer, 1998;
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Diep, Havarstein, & Nes, 1996; Diep, Straume, Kjos, Torres, & Nes, 2009). Their

activity depends, by definition, on the complementary action of two different peptides,

where their cationic nature is essential, facilitating the initial contact between

bacteriocins and the negatively charged membranes via electrostatic interactions (Diep,

et al., 2009).

Finally, plantaricin A, one-peptide bacteriocin without post-translational

modifications, is included in subclass IIc (Diep, et al., 2009). The antimicrobial

spectrum of plantaricin A is relatively narrow, comprising mainly different

Lactobacillus species (Lactobacillus casei, Lactobacillus sakei, Lactobacillus

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viridescens and L. plantarum). The antimicrobial activity of plantiricin A was 10-100

fold lower than plantiricins EF and JK (Anderssen et al., 1998).

3.3. Biosynthesis and mode of action of bacteriocins

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According to Drider, et al. (2006), at least four genes are needed for the

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production and secretion of bacteriocins. In particular, they are (i) the structural

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bacteriocin gene, encoding a prebacteriocin; (ii) the immunity gene, encoding an

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immunity protein that protects the bacteriocin producer from its own bacteriocin; (iii) a

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gene encoding an ABC (ATP-binding cassette) transporter necessary for secretion; and

(iv) a gene encoding an accessory protein of unknown function. Bacteriocins can result
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from the expression of a gene located at the chromosome (plantaricin ST31 for

example) or at a plasmid (plantaricin 423 for example) (Todorov, 2009; Reenen, Dicks,
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& Chikindas, 1998; Todorov et al., 1999). However, when two bacteriocins are
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produced by a strain one can be chromosomal, such as carnobacteriocin BM1, and the
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other from the plasmid, such as carnobacteriocin B2 (Todorov, 2009). Of course, there
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are situations in which two or more bacteriocins can only originate either from the

chromosome or plasmid (Todorov, 2009).

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Most class II bacteriocins are synthesized primarily in the form of a pre-peptide

or a biologically inactive pre-bacteriocin. This compound contains a sequence from 18

to 27 amino acids presenting two glycines at the N-terminus. This sequence has the

function of preventing the bacteriocin to be activated inside the producer cell and serves

as recognition signal for the transport system involving the ABC transporter proteins

and accessory protein (Nes, Diep, Havarstein & Brurberg, 1996; Savadogo, Ouattara

Bassole, & Traores, 2006). The two glycines present in the sequence are responsible for

the recognition by the pre-bacteriocin transport system (Moll, Konings & Driessen,

1999). After recognizing the pre-peptide, the leader amino acid sequence of bacteriocin

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is removed and then the active peptide/ bacteriocin is secreted into the extracellular

medium (Ehrmann, Remiger, Eijsink, & Vogel, 2000).

Regarding the mode of action, different mechanisms have been proposed for

bacteriocins. Such mechanisms directly depend on factors related to bacterial species

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and their growth conditions, bacteriocin dose employed and purification degree (Parada,

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et al., 2007). In particular, these mechanisms can promote a bactericidal effect, with or

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without cell lysis, or bacteriostatic, inhibiting cell growth (Cintas et al., 2001). Usually,

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pore formation – which results in the variation of the cytoplasm membrane potential due

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to the hydronium ion exchanging between the inner and outer membrane surfaces - is

the main mechanism by which most of bacteriocins from LAB exert their antibacterial
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effect (Ghrairi et al., 2012). In class II, this mechanism is triggered when bacteriocin

binds to a protein-receptor on the cell membrane of the target bacteria, although some
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author believe that such protein-receptor does not appear to be essential for binding
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(Chen, Shapira, Eisenstein, & Montville, 1997; Jack, Tagg, & Ray, 1995).
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Although it has been reported that bacteriocins show a bactericidal mode of

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action centered against homologous species, some of them have similar activity against

food-borne pathogens; including the bacteriocin produced by P. acidilactici M that

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inhibits a large number of bacteria, such as S. aureus, L. monocytogenes, C. perfringens,

Bacillus coagulans, B. cereus, and A. hydrophila; or the bacteriocin sakacin C2

produced by L. sakei C2a that inhibits many Gram-positive and Gram-negative bacteria

(Hu, Zhao, Zhang, Yu, & Lu, 2013).

Nevertheless, the antimicrobial activity of bacteriocins is unstable and

inconsistent, as it depends on the chemical and physical conditions of foods. There are

factors that can interfere on the bacteriocin production by LAB such as unsuitable

process conditions (pH, temperature, nutrients, among others), spontaneous cell loss on

producing bacteriocin, infection of the cell by phage and the presence of competitive

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microorganisms in the medium (Schillinger, Geisen, & Holzapfel, 1996). Besides, the

bacteriocin effectiveness would also be affected by the presence of bacteriocin-resistant

microorganisms, enzymes (like proteases), occurrence of oxidation-reduction reactions,

interaction with components of the food formula (fats, proteins, preservatives, pH, for

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instance) and diffusion restraints due to high salt concentration. Additionally, it can be

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influenced by the presence of nitrate and nitrite and low water activity, which can lead

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to inadequate distribution of the bacteriocin throughout the food product (Schillinger et

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al., 1996; Alves, Martinez, Lavrador, & De Martinis, 2006).

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According to Kristo, Koutsoumanis, and Biliaderis (2008), bacteriocins can

present a higher effectiveness when added into films and not directly incorporated to the
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product. In fact, the production of bacteriocin by L. plantarum was higher in cellulose

derivative films when compared with protein films (Sánchez-González, Saavedra, &
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Chiralt, 2013).
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The mode of action of plantaricin C, a bacteriocin produced by strains of L.

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plantarum, depends on the target microorganism, showing different behavior against

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Gram-positive bacteria (González et al., 1994). Thus, this bacteriocin showed a

bactericidal mode of action, with absence of concomitant or subsequent cell lysis, after
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being added to L. sake CECT 906, reducing 50% the viability of exponentially growing

cultures after 1h without absorbance reduction. Meanwhile, it was observed that a

bacteriolytic mode of action against Lactobacillus fermentum LMB 13554 decreases the

viability of culture to 0.6% in only 5 min, accompanied by a drastic lowering in the

optical density. Additionally, it showed complete lysis against Lactobacillus delbrueckii

subsp. bulgaricus LMG 13551 and almost 100% viability reduction without apparent

decrease in optical density using Lactobacillus helveticus LMG 13555 and Leuconostoc

mesenteroides subsp. cremoris NCDO 543. According to these authors, the lytic effect

of plantaricin C was not observed with most LAB bacteriocins being potentially useful

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in accelerated food processing, releasing the enzymes contained in the starters into their

substrates.

Other plantaricins show a broader antimicrobial spectrum inhibiting food-borne

pathogens in addition to the closely related Lactobacilli species. For example, partially

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purified plantaricin 163, produced by L. plantarum 163 isolated from traditional chinese

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fermented vegetables, inhibits Gram-positive bacteria (S. aureus, L. monocytogenes,

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Bacillus pumilus, B. cereus, Micrococcus luteus, Lactobacillus thermophilus, and

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Lactobacillus rhamnosus) and Gram-negative bacteria (E. coli, P. aeruginosa, and

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Pseudomonas fluorescens). However, is unable to show antimicrobial activity against

fungi such as Penicillium notatum, Aspergillus niger, and Rhisopus nigricans (Hu et al.,
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2013). Plantaricin LP84 showed activity against Gram-positive, Gram-negative, food-

borne pathogenic, and spoilage bacteria (Suma, Misra, & Varadaraj, 1998). Plantaricin
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UG1 inhibits strains of the genera Lactobacillus and Lactococcus, in addition to food-
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borne pathogens such as L. monocytogenes, B. cereus, C. perfringens, and Clostridium

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sporogenes (Enan, et al., 1996). Meanwhile, plantaricin MG presented inhibitory

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activity against Gram-positive and Gram-negative bacteria including L. monocytogenes,

S. aureus, Salmonella typhimurium, and E. coli (Gong, Meng, & Wang, 2010).
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4. L. plantarum

L. plantarum is one of the most widespread species of the genus Lactobacillus

and is being widely used in food-related technologies (Brinques, Peralba, & Ayub,

2010; Sauvageau et al., 2012). This microorganism is a facultative heterofermentative

LAB (Group II) (Bove et al., 2012a; Siezen & van Hylckama Vlieg, 2011). It is acid

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tolerant and considered as a safe microorganism (Generally Regard as Safe – GRAS)

(Brinques et al., 2010). As it is a heterogeneous species, it is closely associated with the

species Lactobacillus pentosus, Lactobacillus paraplantarum and, more recently,

Lactobacillus fabifermentans (Parente et al., 2010; Siezen & van Hylckama Vlieg,

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2011). This relationship was identified when more than 99% of their rRNA presented

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identical sequences, suggesting high phenotypic and genotypic similarity between

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species (Parente et al., 2010).

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L. plantarum is the most important and versatile species of the group, and can be

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found as part of the microbiota of starchy foods and cereals, meats, dairy products,

vegetables, fruits and drinks (Ricciardi et al., 2012). According to Todorov et al. (2011),
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different strains have been isolated from various niches, such as fermented milk, cheese,

fermented cucumber, fermented olives, pasta, pineapple, grapefruit juice, sorghum beer
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and barley, molasses, boza, kefir and amasi. These strains have proven to be able to
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survive gastric transit and colonize the intestinal tract of humans and other mammals,
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and it is considered as a member of the natural microbiota of these niches (Kleerebezem

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et al., 2003; Mathara et al., 2008). Some authors report that L. plantarum can adapt to

various niches due to its ability to ferment a wide range of carbohydrates (Prins et al.,
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2010; Brinques, et al., 2010; Todorov, 2008).

Furthermore, L. plantarum has traditionally been used in starter cultures in food

fermentations (Bove et al., 2012b) and also as an ingredient for probiotic foods, such as

the L. plantarum 299v strain, which is widely marketed (Siezen & van Hylckama Vlieg,

2011).

4.1. Bioproduction of bacteriocins from L. plantarum

Several studies have been focused on the optimization of culture medium and

growth conditions in order to increase the production of bacteriocins by L. plantarum.

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There are many works describing the production of bacteriocins by strains of L.

plantarum, namely: L. plantarum ST194BZ (Todorov & Dicks, 2005), L. plantarum

ST13BR (Todorov, Van Reenen, & Dicks, 2004b), L. plantarum ST414BZ (Todorov &

Dicks, 2006b), L. plantarum ST664BZ (Todorov & Dicks, 2006b), L. plantarum

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ST23LD (Todorov & Dicks, 2006a), L. plantarum ST341LD (Todorov & Dicks,

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2006a), L. plantarum AMA-K (Todorov, Nyati, Meincken, & Dicks, 2007a), L.

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plantarum ST26MS, L. plantarum ST28MS and L. plantarum ST32 (Todorov,

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Gotcheva, Dousset, Onno, & Ivanova, 2000; Todorov, Powell, Meincken, Witthuhn, &

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Dicks , 2007b; Todorov, Van Reenen, & Dicks, 2007c; Todorov, 2008).

Despite of the great number of studies carried out until this moment, there is
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little knowledge about the growth conditions required for optimal production of

bacteriocins by L. plantarum, and an ideal fermentation process have not been

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established (Todorov, Van Reenen, & Dicks, 2004b). It is known that cell growth of
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Lactobacillus is directly influenced by the conditions of pH, temperature, medium

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composition, aeration rate, among other factors. Since LAB are quite demanding on
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nutritional requirements, a rich medium is extremely necessary for good growth

(Brinques, et al., 2010).

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4.2. Bacteriocins produced by L. plantarum

A wide variety of bacteriocins produced by different L. plantarum strains have

been isolated and described. Table 1 shows some examples of bacteriocins produced by

L. plantarum isolated from various fermented products, their respectively biochemical

features and some genetic information when were available. The examples are detailed

as follows:

 Meat:

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Various bacteriocinogenic strains of L. plantarum have been isolated from

sausages obtained from different manufacturers under distinct ripening times (Garriga,

Hugas, Aymerich, & Monfort, 1993).

Enan et al. (1996) isolated an antimicrobial substance produced from

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Lactobacillus plantarum UG1 obtained from dry sausage. This substance was capable

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of inhibiting other strains of Lactobacillus and Lactococcus and some pathogenic

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strains, such as L. monocytogenes, B. cereus, C. perfringens and C. sporogenes. This

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antimicrobial compound was characterized as a bacteriocin and named plantaricin UG1.

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This is a single-peptide with molecular mass between 3.0 and 10.0 kDa and its

production appeared to be chromosomally encoded. In addition, a LAB isolated from

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Italian sausages produced a bacteriocin, called plantaricin 35d (MW 4.5kDa), having

high antimicrobial activity against food pathogens (S.aureus, L.monocytogenes and A.

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hydrophila) (Messi et al., 2001).

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Sausage has been employed as a continuous source of bacteriocinogenic

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cultures. For instance, Kanatani and Oshimura (1994) reported the production of
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plasmid encoded bacteriocin called plantacin 154 with molecular mass about 3.0 kDa or

less, produced by L. plantarum LT154 strain isolated from dry sausage. Meanwhile
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Rekhif et al. (1995) obtained plantaricin SA6 from L. plantarum SA6, an isolate from

fermented sausage. This is a single-peptide with molecular mass about 3.4 kDa and

informations about genetic determinants were not mentioned.

A study by Todorov et al. (2010) characterized bacteriocins produced by L.

plantarum ST202Ch and ST216Ch strain isolated from beloura or chorizo, a traditional

Portuguese product made of pork meat. The chromosomally encoded single peptide

bacteriocins were denominated bacST202Ch and bacST216Ch with molecular mass

estimated to be 3.5 and 10 kDa, respectively, and were able to inhibit the growth of

various Gram-positive and Gram-negative microorganisms considered deteriorative of

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meat products. The genes encoding bacteriocin ST202Ch were found identical to that

reported for the structural gene encoding pediocin PA-1 (pedA, pedB, pedC, and pedD).

 Fish:

Noonpakdee et al. (2009) isolated the L. plantarum PMU 33 strain from som-

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fak, a Thai product made of fish with low salt contents. The bacteriocin purified and

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characterized from the culture supernatant consisted of two peptides with the molecular

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masses of 3.2 and 3.0 kDa. The molecular mass of this two-peptide bacteriocin was

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nearly identical to that of two-peptide plantaricin W (Plw) which consists of two

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peptides Plwα and Plwβ. The genes encoding these two peptides amplified by PCR with

Plw gene specific primer showed identical sequences to Plwα and Plwβ. This
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bacteriocin was able to inhibit a large number of Gram-positive microorganisms

considered pathogens and food spoilage microorganisms, such as L. monocytogenes, B.

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cereus, E. faecalis and S. aureus.

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Fricourt et al. (1994) isolated the L. plantarum BF001 strain from the flesh of
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processed and refrigerated catfish. This strain was able to produce an antimicrobial
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substance designated plantaricin F, a single-peptide with molecular mass between 0.4

and 6.7 kDa. It was active against some bacteria of the genus Lactobacillus,
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Lactococcus, Listeria, Micrococcus, Leuconostoc, Pediococcus, Staphylococcus,

Streptococcus, Salmonella and Pseudomonas.

 Fruits and vegetables:

Two bacteriocins, ST28MS and ST26MS, produced by different L. plantarum

strains, were isolated from molasses and partially characterized. Both bacteriocins with

molecular masses estimated to be 5.5 and 2.8 kDa, respectively, showed unusual

antimicrobial activity against Gram-negative bacteria, including P. aeruginosa, E. coli

and A. baumanii. No plasmids were recorded for strains ST28MS and ST26MS,

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suggesting that these bacteriocins are chromosomally encoded (Todorov & Dicks,

2004a).

L. plantarum C-11 isolated from fermented cucumbers (Daeschel et al., 1990)

produced bacteriocins such as plantaricin EF and plantaricin JK (Anderssen et al.,

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1998). Plantaricin A, which was previously incorrectly identified as the bacteriocin

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responsible for the antimicrobial activity of L. plantarum C11, induces the production

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of the bacteriocins mentioned above (Meyer, Larsen, Sletten, Daeschel, & Nes, 1993).

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Díaz et al. (1993) identified two bacteriocins produced by L. plantarum LPC010

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isolated from fermented green olives. These antimicrobial substances were named

plantaricin S (2.5 kDa), which is produced during the logarithmic growth phase, and
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plantaricin T (molecular mass not determined), produced when the microorganism

reaches the stationary growth phase. The genetic determinants for plantaricin S and T
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production do not appear to be plasmid encoded.

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Todorov et al. (2011) isolated the L. plantarum ST16Pa strain from papaya
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(Carica papaya). The bacteriocin produced by this strain, named ST16Pa (6.5 kDa),
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showed activity against different species of the genus Enterobacter, Enterococcus,

Lactobacillus, Pseudomonas, Streptococcus, Staphylococcus, and some genres of

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Listeria spp. No information was given about genetic determinants.

Plantaricin 163 is a novel bacteriocin produced by L. plantarum 163 isolated

from traditional Chinese fermented vegetables by Hu, et al. (2013). This bacteriocin

showed a broad-spectrum inhibitory activity not only against LAB but also against other

Gram-positive and Gram-negative bacteria including S. aureus. L. monocytogenes, B.

pumilus, B. cereus, M. luteus, L. thermophilus, L. rhamnosus, E. coli, P. aeruginosa,

and P. fluorescens. The physicochemical studies of this bacteriocin (3.5 kDa) are in

agreement with the characteristic features of antimicrobial peptides, thus indicating the

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potential value of plantaricin 163 as a biopreservative in the food industry (Hu, et al.,

2013). No information was given about genetic determinants.

Other bacteriocins are plantaricin C19 produced by L. plantarum C19 isolated

from fermented cucumbers (Atrih, Rekhif, Moir, Lebrihi, & Lefebvre, 2001);

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plantaricin NA, produced by L. plantarum sp. isolated from vegetable origin (Olasupo,

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1998); plantaricin-149, produced by L. plantarum NRIC 149 isolated from pineapple

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(Kato et al., 1994); and plantaricin D produced by L. plantarum BFE 905 isolated from

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“Waldorf” salad (Franz, Du Toit, Olasupo, Schillinger, & Holzapfel, 1998).

 Milk-based products: NU
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Todorov et al. (2007a) reported the production of bacteriocin AMA-K by the L.

plantarum AMA-K strain isolated from amesi, a product traditionally made of

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fermented milk that is consumed in different regions of Southern Africa, including

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Zimbabwe, South Africa and Lesotho. The bacteriocin AMA-K (2.9 kDa) inhibited the
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growth of Enterococcus spp., E. coli, K. pneumoniae and Listeria spp. No information

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was given about genetic determinants.

In 1994, González et al. reported the production of plantaricin C by L.

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plantarum LL441 strain isolated from Cabrales cheese. The bacteriocin showed

bactericidal activity, followed by, in some cases, cell lysis.

Xie et al. (2010) demonstrated the presence of pediocin LB-B1 (single-peptide

with molecular mass estimated between 2.5 and 6.5 kDa), which was produced by L.

plantarum LB-B1 isolated from koumiss, a traditional Chinese fermented dairy product.

In particular, this bacteriocin was active against Listeria, Lactobacillus, Streptococcus,

Enterococcus, Pediococcus and E. coli strains. The genetic determinants for pediocin

LB-B1 production appeared to be plasmid-encoded.

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Powell et al. (2007) isolated L. plantarum ST8KF from kefir, a carbonated

refreshing drink, and its bacteriocin (single-peptide with molecular mass about 3.5 kDa)

showed activity against different microorganisms including L. casei, L.salivarius, L.

curvatus and L. innocua. No information was given about genetic determinants.

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Other bacteriocins include plantaricin MG (molecular mass about 2.1 kDa)

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produced by L. plantarum KLDS1.0391 and isolated from “Jiaoke”, a traditional

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fermented cream from China by Gong, et al. (2010), being active against Gram- positive

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and Gram-negative bacteria including L. monocytogenes, S. aureus, S. typhimurium and

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E. coli and plantaricin LC74 (single-peptide with molecular mass about 5 kDa),

produced by L. plantarum LC74 isolated from crude goat’s milk. This bacteriocin
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showed a narrow spectrum of activity against several strains of mesophilic lactobacilli

including L. plantarum, L. brevis and Lactobacillus biichneri (Rekhif, Atrih, &

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Lefebvre, 1994).
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 Cereals:
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Todorov et al. (2004b) reported the production of the bacteriocin ST13BR

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(single-peptide with molecular mass about 10 kDa) by L. plantarum ST13BR strain

isolated from barley beer, a traditional drink made of fermented corn, barley, soy flour
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and sugar, produced in South Africa. This bacteriocin was effective against L. casei, P.

aeruginosa, E. faecalis, K. pneumoniae and E. coli. No information was given about

genetic determinants.

Todorov & Dicks (2005) described the isolation of the L. plantarum ST194BZ

strain from boza, a fermented beverage typically from the eastern Balkan countries. It is

one of the most traditional drinks produced by fermentation of different cereals with the

use of yeast and LAB. The ST194BZ strain was able to produce two types of

bacteriocins, termed as ST194BZα (3.3 KDa) and ST194BZβ (14 kDa), being active

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against a broad range of pathogens and spoilage bacteria including E. faecalis, E. coli,

E. cloacae and P. aeruginosa. No information was given about genetic determinants.

Reenen et al. (1998) isolated the L. plantarum 423 strain from sorghum beer.

This strain produced the bacteriocin plantaricin 423 (single-peptide with molecular

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mass about 3.5 kDa), which is capable of inhibiting a wide range of microorganisms,

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such as B. cereus, C. sporogenes, E. faecalis, Listeria spp. and Staphylococcus spp.

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DNA hybridization studies have shown homology between the plasmid DNA of L.

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plantarum 423 and the pediocin PA-1 operon. This suggests that plantaricin 423 is

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plasmid-encoded.

Additionally, the production of other bacteriocins has been described, including

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plantaricin K, produced by L. plantarum DK9 isolated from “fufu”, a fermented cassava

product (Olukoya, Tichaczek, Butsch, Vogel, & Hammes, 1993); plantaricin ST31,
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produced by L. plantarum ST31 isolated from sourdough (Todorov et al., 1999) and
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plantaricin KW30, produced by L. plantarum strain KW30 isolated from fermented corn
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(Kelly, Asmundson, & Huang, 1996).

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4.3. Applications of L. plantarum and its bacteriocins

A wide variety of benefits associated with the use of L. plantarum as potential

probiotic has been reported in literature (Ningegowda & Gurudutt, 2012; Zago et al.,

2011). In general, these studies reported that this LAB has been used to enhance

intestinal barrier function and improve symptoms of irritable bowel syndrome

(Anderson, Cookson, MacNabb, Kelly, & Roy, 2010; Ducrotté, Sawant, & Jayanthi,

2012). According to Axling et al. (2012), the use of L. plantarum can affect gut

microbiota, lipid metabolism and inflammation in high-fat fed mice. Another example

of health benefits was reported by Nguyen, Kang, and Lee (2012). In this study, L.

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plantarum PH04 was effective in cholesterol-lowering activities. On the other hand,

Gallego et al. (2011) reported that the consumption of probiotic L. plantarum improves

the general health status in older people.

Several substances have been produced by different L. plantarum strains isolated

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from different food sources. These strains of bacteriocinogenic L. plantarum are

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naturally present in these products, contributing to the improvement of organoleptic

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characteristics and playing an essential role in their biopreservation (Todorov, 2009).

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Biopreservation is a technique used to prolong food shelf life and improve

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safety through the use of protective microbiota and/or its antimicrobial peptides, such as

bacteriocins (Schillinger et al., 1996). The application of bacteriocins can help to reduce
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the use of chemical preservatives and/or the intensity of heat treatment, as well as

prevent other physical treatments, satisfying the demand for "fresh", “ready-to-eat”
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foods with few preservatives (Todorov, 2009).

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The bacteriocins produced by L. plantarum or even the strain itself are used in
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different situations. Penteado et al. (2007) conducted a study by inoculating bacteriocins

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from L. plantarum in silage of mombaça grass, with the purpose of improving the

fermentation profile according to the values of pH, NH3, lactic acid and acetic acid,
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favoring the development of LAB and lowering dry matter losses.

Campagnol, Fries, Terra, Santos, and Furtado (2007) produced a starter culture

with a L. plantarum strain in the culture medium of porcine plasma and verified the

feasibility of its application in salamis. As a result, salamis made with a L. plantarum

starter culture promoted a higher microbiological safety to salamis, as well as a

significant improvement of their flavor.

In the therapeutic field, the topical application of lactobacilli is receiving

attention due to the prevention of superficial skin and burn wound infections.

Brachkova et al. (2011) studied the properties of L. plantarum immobilized with

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calcium alginate films and investigated the antibacterial activity of these films in a

model burn wound in rats. The study showed a significant decrease in the number of

colonies of P. aeruginosa (one of the most frequently isolated pathogens in chronic

infections, which was used to test the activity of L. plantarum), suggesting that the

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immobilization of L. plantarum by calcium alginate films may be a possible

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intervention for the prevention of infections caused by burns. Also in this context,

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Ramos et al. (2012) investigated the effects of L. plantarum supernatants on pathogenic

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properties of P. aeruginosa, such as adhesion, viability, virulence factors, biofilm

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formation, and quorum sensing signal expression. In this study, L. plantarum

supernatants were able to inhibit pathogenic properties of P. aeruginosa by a quorum

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quenching mechanism. Also, the antipathogenic properties, along with the

immunomodulatory, tissue repair, and angiogenesis properties in the L. plantarum

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supernatants, make them an attractive option in infected chronic wound treatment.

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O’Shea et al. (2012) reported the problem related to the odor of pig manure.
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Conventional dietary strategies to reduce this odor can be costly, prevent nutrient
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digestibility, and receive varying responses. Alternatively, the authors proposed the use

of L. plantarum in the diet of pigs, with or without supplementation of inulin, in order to

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reduce the manure odor without compromising the nutrient digestibility. It was found

that dietary supplementation of pigs with L. plantarum, with or without inulin, reduces

the manure odor.

4.4. Purification of bacteriocins produced by L. plantarum

Different strategies for the extraction of bacteriocins produced by L. plantarum

from cultivation broths, and further purification to final products have already been

described in literature (Atrih et al., 2001; Gong et al., 2010; Hata, Tanaka, & Ohmomo

et al., 2010; Müller, Carrasco, Tonarelli, & Simonetta, 2009; Smaoui et al., 2010;

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Todorov, Velho, & Gibbs, 2004c; Zhu, Zhao, Sun, & Gu, 2014). Purification methods,

including salinization, solvent extraction, ultrafiltration, adsorption-desorption, ion

exchange chromatography and High-Performance Liquid Chromatography (HPLC), are

the most common techniques (Parada et al., 2007). Other alternative purification

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methods include salting-out, gel filtration, or Reverse-Phase High-Performance Liquid

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Chromatography (RP-HPLC) (Hu et al., 2013).

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Some purification strategies with the respective specific activity and purification

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folds are shown in Table 2. All of them led to high levels of purification indicating the

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high effectiveness degree of them.

The literature related to the purification of bacteriocins is vast, comprehending

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several types of protocols. However, all protocols involve the use of HPLC technique as

the final purification step (Hata et al., 2010; Smaoui et al., 2010; Zhu et al. 2014). Some
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authors employed the RP-HPLC technique, a variant of HPLC in which a hydrophobic

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stationary phase is employed, favoring, of course, the elution of polar molecules (Atrih
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et al., 2001; Gong et al., 2010; Müller et al., 2009). It must be borne out that by using
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the RP-HPLC technique as the final step, less purification procedures are required, thus

decreasing the overall cost of the purification process.

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Recently, it was proposed a new procedure for purification of macromolecules,

including bacteriocins, based on the liquid-liquid extraction for aqueous two-phase

micellar systems (ATPMS). This method can be applied for extracting bacteriocins

directly from the fermented medium, leading to a simplification on the overall

purification protocol of bacteriocins (Jozala, Lopes, Novaes, Mazzola, & Pessoa-Jr,

2012; Liu, Nikas, & Blankschtein, 1996; Molino et al., 2014).

5. Conclusion

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Nowadays, consumers tend to seek fresh and natural products, avoiding

processed products containing chemical additives. Thus, many researchers began to

look for natural and effective preservatives. The use of bacteriocins seems to be a great

alternative, as they present activity against a wide range of food-borne pathogens and

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spoilage microorganisms.

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Several substances have been isolated from various L. plantarum strains found

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in different niches, such as meat, fish, dairy products, fermented vegetables, cereals and

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fruits. Their applicat ion can be performed successfully in food-related fermentations,

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ensuring not only the organoleptic characteristics, but also contributing to increase the

shelf life and safety of the final product.

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Although there are many studies on the production of bacteriocins by L.

plantarum, until this moment, there are no reports in literature describing the existence
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of a bacteriocin from L. plantarum, such as nisin and pediocin, available in the market.
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This fact could be due to the lack of an efficient or the low-cost purification strategy,
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which allows the bacteriocin produced by this microorganism to be commercially

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available. Therefore, future studies might be directed at the development of efficient and

low-cost purification protocols.

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Acknowledgements

We are grateful to FAPESP (Fundação de Amparo à Pesquisa do Estado de São

Paulo) for the financial support of this work (processes numbers 2013/19997-5,

2012/23340-9 and 2013/12713-1).

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Table 1

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IP
Some examples of bacteriocins produced by Lactobacillus plantarum isolated from various ecological niches

CR
Isolation Strain name Bacteriocin Biochemical Genetic Some pathogens inhibited Reference
niche produced features information

US
L. plantarum UG1 plantaricin UG1 Single-peptide with Chromosomally L. monocytogenes, B. cereus; C. Enan et al., 1996

N
molecular mass encoded perfringens and C. sporogenes
Meat

MA
between 3.0 and 10.0
kDa

D
TE
L. plantarum 35d plantaricin 35d Single-peptide with Information S. aureus, L. monocytogenes and Messi et al, 2001
molecular mass about genetic A. hydrophila

P
estimated to be 4.5 determinates not
CE
kDa shown
L. plantarum plantacin 154 Single-peptide Plasmid- Enterococcus faecalis, Kanatani &
AC

LT154 peptide with encoded Bacillus spp., Staphylococcus spp. Oshimura, 1994
molecular mass and S. typhimurium
estimated to be 3.0
kDa or less
L. plantarum SA6 plantaricin SA6 Single-peptide Information L. plantarum, Lactobacillus brevis, Rekhif, Atrih, &
peptide with about genetic Leuconostoc spp and Listeria grayi Lefebvre, 1995
molecular mass determinates not
estimated to be 3.4 shown
kDa

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L. plantarum bas ST202Ch Single-peptide with Chromosomally Enterococcus faecium, E. coli, L. Todorov, Ho, Vaz-
ST202Ch and bac ST216 molecular mass encoded monocytogenes, Pseudomonas Velho, & Dicks,

T
ST216Ch estimated to be 3.5 spp. and S. aureus. 2010

IP
and 10 kDa,

CR
respectively
L. plantarum PMU plantaricin W Two-peptide (α and Chromosomally L. monocytogenes, B. cereus, S. Noopakdee et al.,

US
33 encoded aureus, E. faecium and E. faecalis. 2009
Fish β-peptide) with
molecular masses

N
estimated to be 3.2

MA
and 3.0 kDa,
respectively

D
L. plantarum plantaricin F Single-peptide with Information S. aureus, S. typhimurum, L. Fricourt, Barefoot,

TE
BF001 molecular mass about genetic monocytogenes and P. aeruginosa. Testin, &
between 0.4 and 6.7 determinates not Hayasaka,1994

P
kDa shown
L. plantarum
ST28MS and
ST28MS and
ST16MS
CE
Single-peptide with
molecular masses
Chromosomally
encoded
L. sakei, S. aureus, E. faecalis, P.
aeruginosa, E. coli and A. baumanii
Todorov & Dicks,
2004a
AC
Fruits and ST16MS estimated to be 5.5
vegetables and 2.8 kDa,
respectively
Lactobacillus plantaricin EF, Two two-peptide with Chromosomally Lactobacillus sp.; Pediococcus sp.; Daeshel,
plantarum C11 plantaricin JK data not shown about encoded Leuconostoc sp. and Streptococcus McKenney, &
and inductor molecular mass sp, McDonald, 1990
factor plantaricin
A

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L. plantarum plantaricin S and Two-peptide, where Possible Propionibacterium sp., Díaz, Sánchez,
LPC010 plantaricin T plantaricin S had chromosomally Clostridium tyrobutyricum and Desmazeaud,

T
molecular mass about encoded E. faecalis Barba, & Piard,

IP
2.5 kDa and the 1993

CR
molecular mass of
plantaricin T was not

US
detected
L. plantarum bacteriocin Single-peptide Information E. faecalis, E. faecium,, L. Todorov et al., 2011

N
ST16Pa ST16Pa peptide with about genetic monocytogenes,, Listeria innocua,,

MA
molecular mass determinates not S. aureus, Streptococcus spp., and
estimated to be 6.5 shown Pseudomonas spp.
kDa

D
TE
L. plantarum 163 plantaricin 163 Single-peptide with Information S. aureus, L. monocytogenes, B. Hu et al., 2013

P
molecular mass about about genetic pumilus, B. cereus,
CE
3.5 kDa determinates not
M. luteus, L. thermophilus, L.
shown
AC
rhamnosus, E..coli, P. aeruginosa
and P. fluorescens

L. plantarum bacteriocin Single-peptide with Information Enteroccus spp., E. coli, Klebsiella Todorov et al.,
Milk AMA-K AMA-k molecular mass about about genetic pneumoniae and 2007a
products 2.9 kDa determinates not Listeria spp.
shown
L. plantarum pediocin AcH Single-peptide with Plasmid- L. monocytogenes Ennahar et al., 1996
WHE92 molecular mass about encoded

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4.5 kDa

T
L. plantarum LB- pediocin LB-B1 Single-peptide with Plasmid- Listeria spp., Lactobacillus spp., Xie et al., 2010

IP
B1 molecular mass encoded Streptococcus spp., Enterococcus

CR
estimated between spp, Pediococcus spp. and E.coli
2.5 and 6.5 kDa

US
L. plantarum BacST8KF Single-peptide with Information L. casei, Lactobacillus salivarus, Powell, Witthuhn,
ST8KF molecular mass about about genetic Lactobacillus curvatus and L. Todorov, & Dicks,

N
3.5 kDa determinates not innocua 2007

MA
shown

L. plantarum bacteriocin Single-peptide with Information P.aeruginosa, E. Faecalis, Todorov, Van

Cereals ST13BR ST13BR molecular mass about about genetic K.pneumonia and E. coli Reenen, & Dicks,

D
10.0 kDa determinates not 2004b

TE
shown
L. plantarum ST194BZ(α) and Two-peptide (α and Information E. faecalis, E. coli, Enterobacter Todorov & Dicks,

P
ST194BZ ST194BZ(β) about genetic cloacae and P. aeruginosa 2005a
CE
β-peptide) with
determinates not
molecular mass
shown
AC
estimated to be 3.3
and 14.0 kDa,
respectively
L. plantarum 423 plantaricin 423 Single-peptide with Plasmid- B. cereus, C. sporogenes, E. Reenen et al., 1998
molecular mass about encoded faecalis, Listeria spp. and
3.5 kDa Staphylococcus spp.

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Table 2. Strategies of purification to obtain bacteriocins produced by Lactobacillus plantarum

T
Bacteriocin Purification steps Specific activity Purification References

IP
(AU/mg) (fold)

CR
Culture supernant; 14.9 1.0
1 – 37.5 1 – 2.5 Zhu et al., 2014
1 – Macroporous resin column;

US
Plantaricin 2 – 369.9 2 – 24.8
ZJ008 2 – Cation exchange chromatography; 3 – 838.7 3 – 56.2

N
4 – 8556.7 4 – 573.1

MA
3 – Gel filtration;

4 - HPLC

D
Culture supernant; 85.5 1 Müller et al., 2009

TE
Plantaricin 1 – 5959.0 1 – 69.6
1 – Sep-Pack cartridges (C18);
2 – 5900.0 2 – 689.5

P
from L.
3 – 506000.0 3 – 5914.6
plantarum LP31 2 – Gel-filtration chromatography; CE
3 – RP-HPLC
AC
Culture supernant; 455.0 1.0
Plantaricin C19 1 – 17808.0 1 – 39.1 Atrih et al., 2001
1 - Release of adsorbed bacteriocin
2 – 409600.0 2 – 900.2
from producing cells;

2 – RP-HPLC

Culture supernant; 0.4 1.0

Plantaricin MG 1 – 5.4 1 – 14.0 Gong et al., 2010
2 – 44.6 2 – 20.0

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1 - Ammonium sulfate precipitation; 3 – 9333.3 3 – 25.2

T
2 - Gel filtration chromatography;

IP
3 – RP-HPLC

CR
Culture supernant; 2083.0 1.0

US
1 – 9904.0 1 - 4.7 Smaoui et al. (2010)
1 - Ammonium sulfate precipitation;
BacTN635 2 – 14310.0 2 – 6.8

N
3 – 146104.0 3 – 70.1
2 – Cintrifugal microconcentrators;

MA
4 – 197368.0 4 – 94.7
3 - Gel filtration;
4 – HPLC

D
Culture supernant; 253.0 1.0

TE
Plantaricin 1 – 1850.0 1 – 7.3 Hata et al., 2010
1 - Ammonium sulfate precipitation;
ASM1 2 – 11900.0 2 – 47.0

P
3 – 20700.0 3 – 81.8
2 - Cation exchenge chromatography;
4 – 10700.0
CE 4 – 42.3
3 – Octyl-Sepharose CL-4B column;
AC

4 – HPLC

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Highlights

T
IP
CR
 Lactic acid bacteria produce peptide bacteriocins such as the plantaricin obtained from Lactobacillus plantarum

US
 L. Plantarum, a facultative heterofermentative lactic acid bacterium, ia one of the most widespread species of the genus Lactobacillus

N
 Plantaricins have been mainly ascribed to Subclasses IIa and IIb, the groups most widely used in food preservation and medicine

MA
 Plantaricins have emerged for biopresevation due to their broad antimicrobial spectrum against food-borne pathogens

D
 Different strategies have been addressed to purify bacteriocins including a final HPLC stage

P TE
CE
AC

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