Lactobacillus spp.
: Lactobacillus plantarum
A Corsetti and S Valmorri, Università degli Studi di Teramo, Mosciano S. Angelo (TE), Italy
ª 2011 Elsevier Ltd. All rights reserved.
This article is a revision of the previous edition article by
A. Corsetti, M. Gobbetti, Volume 3, pp 1501–1507, ª 2002,
Elsevier Ltd.
Taxonomy, Identification, and Genetics
of the Species
Lactobacillus plantarum is a Gram-positive, nonmotile,
non-spore-forming, microaerophilic, and mesophilic bac-
terium with growth occurring at 10–15  C but not
at 45  C. Cells are straight rods with rounded ends,
0.9–1.2  3.0–8.0 mm, occurring singly, in pairs, or in short
chains. It has been found that under special conditions, a few
strains of Lb. plantarum possess true catalase and manganese-
containing pseudocatalase activities. Some strains also
exhibit nitrate- and hematin-dependent nitrite reductases.
The cell wall contains either ribitol or glycerol
teichoic acid although some strains have an unusual tei-
choic acid. Peptidoglycan of the cell wall is of the
meso-diaminopimelic acid (DAP) type.
On the basis of 16S rRNA-based phylogeny, recently
supported by whole-genome DNA, DNA–RNA hybridi-
zation, and GC content studies, Lb. plantarum belongs to
the Lactobacillus casei–Pediococcus group; DNA–DNA
hybridization studies have demonstrated that Lb. plantarum
forms two genomically related groups, in their turn sepa-
rated by other streptobacteria and lactobacilli containing
DAP in the cell wall. Even if simple methods like random
amplified polymorphic DNA-polymerase chain reaction
(RAPD-PCR) have been used for both strain typing and
differentiation from similar Lactobacillus spp., a polyphasic
strategy based on physiological tests and more than one
molecular technique is necessary for the correct species
designation.
Lactobacillus plantarum, Lactobacillus pentosus, and
Lactobacillus paraplantarum form a closely related group
known as the ‘Lactobacillus plantarum group’, the species
of which cannot be distinguished from each other by 16S
rRNA gene sequence analysis because of 99.7–99.9%
sequence similarity; the species of the group also show a
very similar fermentation pattern. Discrimination among
these species can be obtained using species-specific PCR
primers based on recA gene and by employing repetitive
element sequence-based polymerase chain reaction
(Rep-PCR) moreover, strain differentiation between
Lb. plantarum and Lb. pentosus could be achieved using
RFLP–PFGE, 16S ARDRA, AFLP using restriction
enzymes HindIII and MseI, as well as ribotyping and
RAPD–PCR. Multilocus sequence typing (MLST)
based on pgm, ddl, gyr B, purK1, gdh, and mutS housekeep-
ing genes can be useful to determine the genetic
relatedness and to discriminate among Lb. plantarum
strains. In recent years, culture-independent methods
have been developed and applied to the direct identifica-
tion of bacteria from many matrices: DGGE and TGGE
are the most useful techniques for this purpose and were
successfully applied to identify Lb. plantarum in cheese and
dairy products.
By means of recA gene sequencing and Southern hybri-
dization with pyr probe on BglI digestion of chromosomal
DNA, Lb. plantarum has been subdivided into subspecies
Lb. plantarum subsp. plantarum and Lb. plantarum subsp.
argentoratensis.
The complete genome of Lb. plantarum WCFS1, a
single colony isolated from Lb. plantarum NCIMB8826
(National Collection of Industrial and Marine Bacteria,
Aberdeen, UK), has been sequenced and deposited in
EMBL database with accession number AL935263.
Lactobacillus plantarum WCFS1 contains a single circular
chromosome of 3 308 274 bp (Figure 1), which can be
considered one of the largest genome sizes known for
lactic acid bacteria; two small cryptic plasmids of 1917
and 2365 bp and one large plasmid of 36 069 bp encoding
genes involved in conjugal plasmid transfer as well as
other functions have also been found. The G þ C content
of the chromosome is 44.5% while the plasmids show a
lower, from 34.3 to 40.8%, G þ C content.
The Lb. plantarum genome contains five rRNA operons
distributed around the chromosome, displaying a very
limited number of sequence polymorphisms; 3502 pre-
dicted protein-encoding genes are present in the
chromosome and putative biological functions could be
assigned to 2120 of the predicted proteins, whereas the
three above-mentioned plasmids contain, in the order of
length, 3, 4, and 43 genes, respectively. Microarray ana-
lysis revealed the presence of two large regions of
flexibility inside the chromosome, which run from 2.70
to 2.85 Mb and from 3.10 to 3.29 Mb; 293 genes, organized
in clusters of 3–6 genes, have been predicted in these
regions, and most of these are involved in sugar transport
111
112 Lactic Acid Bacteria | Lactobacillus spp.: Lactobacillus plantarum

Figure 1 Genome-atlas view of the Lactobacillus plantarum WCFS1 chromosome, with the predicted origin of replication at the top.
The circles show (from outer to inner) (1) positive strand ORFs (red), (2) negative strand ORFs (blue), (3) GC skew (green), (4) G þ C
content (black), (5) prophage-related functions (green) and IS-like elements (purple), and (6) rDNA operons (black) and tRNA encoding
genes (red). The GC% and GC skew (C  G)/(C þ G) were calculated in a window of 4000 nt, in steps of 75 nt. The G þ C percentage was
plotted as the number of G þ C nucleotides in the plus strand divided by the window size, that is, (G þ C)/4000; the lowest and highest
values are 30.8 and 51.8%, respectively. The upper and lower values of the GC skew are 0.22 and 0.27, respectively. Reproduced from
Kleerebezem M, Boekhorst J, van Kranenburg R, et al. (2003) Complete genome sequence of Lactobacillus plantarum WCFS1.
Proceedings of the National Academy of Sciences of the United States of America 100(4): 1990–1995.
www.pnas.org_cgi_doi_10.1073_pnas.0337704100. Copyright (2009) National Academy of Sciences, U.S.A.

(phosphotransferase system (PTS)) and catabolism.
Moreover, many genes in this region show an unusual
base composition compared with the rest of the genome,
which could be an indicator of horizontal gene transfer.
On the basis of these findings, it has been hypothesized
that this part of the Lb. plantarum genome represents a
‘lifestyle adaptation island’ of the species to different
environmental niches.
The type strain of the species is Lb. plantarum ATCC
14917.
Metabolism and Enzymes
Lactobacillus plantarum is included in the group of faculta-
tively heterofermentative lactobacilli; hexoses are
converted almost entirely to lactic acid via the Embden–
Meyerhof–Parnas pathway, while pentoses are converted to
lactic and acetic acids via the 6-phosphogluconate/phos-
phoketolase pathway by the induction of phosphoketolase.
Both L- and D-lactic acid are produced by NAD-dependent
lactate dehydrogenases (LDHs); amygdalin, arabinose, cel-
lobiose, fructose, galactose, glucose, gluconate, lactose,
maltose, mannitol, mannose, melibiose, raffinose, ribose,
salicin, sorbitol, sucrose, and trehalose are used as carbon
and energy sources. Sugars are usually transported into the
cell by permeases and are phosphorylated in the cytoplasm;
the presence of 25 complete phosphoenolpyruvate–
phosphotransferase (PEP–PET) sugar transport systems,
as deduced by the complete sequence of the chromosome
of Lb. plantarum WCFS1, reflects the adaptative behavior of
the species.
Besides lactic acid, many secondary end products can be
produced from pyruvate (Figure 2), the nature of which
depends on the strain and culture parameters. For example,
in the presence of oxygen, glucose is converted to acetate
from pyruvate via acetylphosphate; the same end product
is obtained in anaerobiosis, but in the presence of a neutral
or alkaline pH or an external electron acceptor (e.g.,
citrate). The formation of acetate via acetylphosphate
increases the synthesis of ATP through acetate kinase
activity. Ethanol too can be produced by some Lb. plantarum
strains under different culture conditions, such as the aero-
bic and anaerobic catabolism of glucose or from external
electron acceptors (e.g., citrate or acetate) during the anae-
robic catabolism of mannitol.
In addition, Lb. plantarum can produce a variety of
secondary end products, such as diacetyl, acetoin, and
 Lactic Acid Bacteria | Lactobacillus spp.: Lactobacillus plantarum 113

Figure 2 Glucose and citrate degradation pathways proposed for the Lactobacillus plantarum wild-type strain (NCIMB8826) and
LDH-defective strain (TF103). (1) LDH; (2) pyruvate formate-lyase; (3) acetaldehde dehydrogenase; (4) alcohol dehydrogenase;
(5) phosphotransacetylase; (6) acetate kinase; (7) pyruvate oxidase; (8) pyruvate decarboxylase; (9) -acetolactate synthase;
(10) -acetolactate decarboxylase; (11) 2,3-butanediol dehydrogenase; (12) nonenzymatic decarboxylation; (13) pyruvate carboxylase;
(14) malate dehydrogenase; (15) fumarase; (16) fumarate reductase; (17) malolactic enzyme; (18) citrate lyase; (19) oxalacetate
decarboxylase; (20) mannitol-1-phosphate dehydrogenase; (21) mannitol-1-phosphatase–enzyme IIMtl. CoA, coenzyme A; 2e,
electrons transferred to an unknown acceptor; Tpp, thiamine PPi. Reproduced with permission from Ferain T, Schanck AN, and
Delcour J (1996) 13C nuclear magnetic resonance analysis of glucose and citrate end products in a ldhL-ldhD double-knockout strain of
Lactobacillus plantarum. Journal of Bacteriology 178: 7311–7315.

2,3-butanediol, which represent key flavor compounds
in many dairy products, from glucose and citrate meta-
bolism via -acetolactate. Other end products from
citrate metabolism can be lactic, acetic, and succinic
acids as well as CO2. Citrate metabolism of Lb. plantarum
is greatly affected by external pH (optimum pH is 4.5)
and by the physiological state of the cells (e.g., growing
or not growing). It has been reported that in growing
cells, metabolism of citrate is higher in cofermentation
with galactose than in the presence of glucose or lac-
tose, whereas in nongrowing cells, metabolism is not
significantly enhanced by the presence of fermentable
sugars. Moreover, citrate is metabolized more rapidly
than sugar by nongrowing cells; on the contrary,
growing cells show a more rapid sugar consumption.
Malic and tartaric acids too are metabolized directly
or in cofermentation with carbohydrates, to produce
CO2, lactic and acetic acids, and other by-products.
Through the malolactic fermentation, Lb. plantarum con-
verts malic acid into lactic acid and CO2, which
represents an important metabolism in the production
of some type of wine.
On the basis of the sequenced genome of Lb. plantarum
WCFS1, metabolic pathways have been reconstructed.
LacplantCyc includes 129 pathways and 704 predicted
reactions involving 670 chemical species and 710
enzymes. It represents the most extensively curated path-
way genome for Gram-positive bacteria.
114 Lactic Acid Bacteria | Lactobacillus spp.: Lactobacillus plantarum

Reconstruction of the metabolic pathways of
Lb. plantarum WCFS1 confirmed the prototrophy of the
species toward folic acid, thiamine, and pyridoxal-5-
phosphate. The same study showed that most pathways
for amino acid biosynthesis are complete. Only arginine,
glutamate, tryptophan, and branched-chain amino acids
(isoleucine, leucine, and valine) are essential for growth
specific intracellular and/or extracellular lipases and
intracellular esterases. An intracellular tributyrin esterase
of 85 kDa and an intracellular lipase of 65 kDa have been
purified and characterized from Lb. plantarum DPC 2739
isolated from Cheddar cheese. Lactobacilli mainly or only
hydrolyze esters containing C4–C6 fatty acids. The ester-
ase activity of some Lb. plantarum strains is highest on
of the species. Auxotrophy for glutamate is caused by
an incomplete TCA cycle and hence no supply of
-ketoglutarate. Synthesis of the tryptophan is regulated
by a feedback mechanism in the presence of other amino
acids such as phenylalanine and tyrosine, and for that
reason, tryptophan is required for growth. The auxotro-
phy for arginine has not yet been clarified.
Presence of glucose increases the conversion of
methionine, and the aminotransferase activity shows vari-
able specificity for the amino group acceptors glyoxylate,
ketoglutarate, oxalacetate, and pyruvate. In Lb. plantarum,
serine is deaminated to pyruvate and ammonia probably
by serine dehydratase. Pyruvate is then catalyzed by
pyruvate–formate lyase to acetate, formate, and CO2;
acetoin is also produced in small quantities from serine.
Lactobacillus plantarum possesses an arginine–deiminase
(ADI) pathway. Through this pathway, citrulline,
ornithine, and ammonia are formed from arginine, driving
an energy supplement for the cell and contributing to pH
homeostasis because of the NH3, which combines with
protons. Moreover, ornithine is a precursor for the
formation of volatile compounds such as 2-acetyl-1-
pyrroline, which represents a key flavor compound in
bread crust. Benzaldehyde is formed from phenylpyruvic
acid, derived by the aminotransferase activity on phenyl-
alanine, in the presence of high levels of Mn2þ, and
contributes to the generation of flavor compound during
cheese ripening. Mn2þ also presents a pseudocatalase
activity, which replaces SOD in a defense mechanism of
the cell against oxygen toxicity.
When compared with other mesophilic lactobacilli,
Lb. plantarum shows a lower proteolytic activity, higher
-naphthyl butyrate, but -naphthyl esters of C16, C18,
and C18:1 fatty acids are also hydrolyzed. In general, lipase
activity of the strains decreases progressively as the length
of the fatty acid chain increases from tributyrin to tripal-
mitin, even if a strain isolated from dry fermented
sausages was found to produce an extracellular lipase
active on triolein. On the other hand, recent studies con-
ducted on strains isolated from salted meat reported that
of 17 strains of Lb. plantarum tested, none was found to
possess lipolytic activity.
Proteins (12) and enzymes (30), mainly hydrolases and
transglycosylases, have been analyzed and are predicted
to be involved in adherence to host components such as
mucin and collagen; this finding suggests that they could
play a role in substrate degradation and can maintain the
growth of Lb. plantarum in different environmental niches.
Antimicrobial Substances and Other
Compounds with Potential Impact on Food
Quality
Besides organic acids, lactic acid bacteria can produce
many inhibitory compounds with activity against other
microorganisms. Bacteriocins, phenyllactic acid, peptides,
and fatty acids are the most active ones and could be of
interest in relation to food biopreservation.
Bacteriocins are ribosomally synthesized proteins or
peptides, often cationic, amphiphilic, and membrane
permeabilizing, that inhibit growth of other bacteria.
They are usually divided into three classes on the basis
of common, mainly structural, characteristics. Class I is
on -casein as on s1-casein, than reported for some
strains. Nevertheless, a high proteolytic activity has
been reported for the strain Lb. plantarum DBPZ1015
during sourdough fermentation, with the formation of
consistent amount of free amino acids. Proteinase and
peptidase activities have been detected in the cell lysates
of Lb. plantarum isolated from cheese, and studies con-
ducted on cell-free extracts of Lb. plantarum
demonstrated higher specific activity toward hydropho-
bic as compared with hydrophilic dipeptides. Peptides are
hydrolyzed by intracellular peptidases of Lb. plantarum,
several of which were characterized at the biochemical
of small, heat-stable peptides containing thioether amino
acids, such as lanthionine, and is hence named lantibio-
tics. Class II is of small, hydrophobic, heat-stable,
nonmodified bacteriocins consisting of either a single
peptide with antilisterial activity (class IIa) or two poly-
peptide chains (class IIb), and also includes other
peptide bacteriocins (class IIc). Class III is of large,
hydrophilic, heat-labile proteins. Bacteriocins produced
from Lb. plantarum, generally indicated as plantaricins,
have been included in both classes I and II. Some exam-
ples are plantaricin C and W (class I); plantaricin C19
and 423 and pediocin AcH (class IIa); plantaricin EF, JK,
and genetic level. S, and NC8 (class IIb); and plantaricin 1.25 (class IIc).
Compared with other cheese-related mesophilic and Production of plantaricins is dependent on pH and tem-
thermophilic Lactobacillus spp., Lb. plantarum shows high perature, and maximum yield is generally obtained at
levels of esterolytic and lipolytic activity related to strain- neutral pH and on incubating the producer strains at
 Lactic Acid Bacteria | Lactobacillus spp.: Lactobacillus plantarum
30  C. Strains of Lb. plantarum that produce bacteriocins
have been isolated from various vegetal and animal
matrices such as cereals, sourdough, wine, meat, and
dairy products. Examples of bacteriocins produced by
Lb. plantarum isolated from dairy products are plantaricin
C, plantaricin TF711, and pediocin AcH.
In recent years, PCR-based methods have been devel-
oped for the identification of bacteriocin-encoding genes;
several genes, including plnA, plnEF, plnJ, plnK, plnJKLR,
plnMNOP, plnABCD, plnEFI, and plnGHSTUV, have
been associated with plantaricin production. Genes
plnEF and plnJ have been reported to encode two-peptide
bacteriocins; the pln locus of bacteriocin produced from
the strains Lb. plantarum C11, NC8, J23, and J51 showed
different patterns of operons involved in transport, secre-
tion of induction factor, and response regulation of the
bacteriocin biosynthesis. It is a general opinion that pro-
duction of several bacteriocins from Lb. plantarum is
quorum-sensing regulated in the presence of a threshold
cell density in the environment, which determines the
secretion of the induction factor.
115
Carbon source and temperature greatly influence EPS
synthesis by Lb. plantarum; compared with glucose, galac-
tose, fructose, and sucrose, lactose is the most efficient
carbon source for EPS biosynthesis by strain EP56. To
optimize the fermentation parameters for EPS production
by Lb. plantarum, statistical and computational methods
such as artificial neuronal networks have been applied.
Functional Activities and Health Benefits
A large number of lactic acid bacteria have been classified
as probiotics. According to the definition adopted by the
World Health Organization, probiotics are ‘‘live micro-
organisms that when administered in adequate amounts
confer a health benefit to the host’’. Probiotics are either
incorporated into functional foods or marketed as lyophi-
lized cells; criteria for selection of a probiotic include its
ability to reach the final destination in a high live number,
adhere to mucus and epithelial cells, and survive at low pH
values and high bile salt concentrations. Susceptibility to
Other nonbacteriocin antimicrobial proteins or peptides
are produced by lactic acid bacteria; Lb. plantarum MiLAB14
produces two cyclic peptides (cyclo(L-Phe-trans-4-OH-L-
Pro) and cyclo(L-Phe-L-Pro)) and one protein, all showing
antifungal activity at a concentration on the order of milli-
gram per milliliter; the same strain produces antifungal
hydroxylated fatty acids active at 10 mg ml–1.
The major antifungal compounds of some Lactobacillus
spp., including Lb. plantarum, are represented by phenyllac-
tic acid (PLA) and 4-hydroxyphenyllactic acid (OH-PLA),
which are derived from the catabolism of phenylalanine;
these compounds showed inhibitory activity against
Fusarium spp., Penicillium spp., and mycelial growth of
Aspergillus niger.
Lactic acid bacteria also produce high amounts and a
great variety of homo- and hetero-exopolysaccharides
(EPSs). These compounds play an important role in the
manufacture of fermented dairy products as safe additives
to improve texture, viscosity, and stability.
The biosynthesis of bacterial EPSs is complex and
unstable, and a large number of genes are involved.
Molecular approaches have been used to characterize
the genes encoding EPSs. The presence of a glycosyl-
transferase gene has been reported for Lb. plantarum.
EPSs are generally produced from Lb. plantarum during
the exponential phase of growth and reach the maximum
amount at the beginning of the stationary phase. Some
strains of Lb. plantarum can produce more than one type of
EPS, differing in terms of molecular mass and sugar compo-
sition. A study carried out on the strain Lb. plantarum EP56
revealed the production of two polymers of 8.5  105 and
4  104 Da, differing in the content of N-acetyl galactos-
amine and rhamnose, respectively.
antibiotics, -galactosidase activity, and ability to utilize
fructooligosaccharides (FOSs) are other important charac-
teristics to evaluate. With regard to this point, it has been
recently reported that different types of FOSs, recognized
as prebiotics, can be fermented by Lb. plantarum because of
the presence of specific transporters and hydrolases;
genetic studies related this ability to the presence of oper-
ons possessing different architecture, which suggests that
these genes were acquired after evolutionary divergence of
the species.
Even though it has been demonstrated that coloniza-
tion of gastrointestinal (GI) tract can be dependent on
the individual, the complete sequence of the genome of
Lb. plantarum WCFS1 can be useful to elucidate many
molecular mechanisms related to the intestinal proper-
ties of this species. The large number of genes encoding
surface proteins (217 predicted proteins) could be
involved in the environmental adaptation of some
strains, because several of those genes show homology
to mucus-binding, aggregation-promoting, and intracel-
lular adhesion proteins. Lactobacillus plantarum strain
299v (DSM 9843) is marketed as a probiotic since 1999;
this strain showed an ability to adhere to human cells in a
mannose-inhibited manner (that is indicative of binding
to a mannosylated cell-bound receptor). Studies per-
formed using DNA microarray analysis on the strain
Lb. plantarum WCFS1 revealed the presence of two
genes involved in mannose adhesion. This finding
could be of interest considering that mannose residues
are commonly found on the surface of eukaryotic cells. A
daily consumption for 3 weeks of Lb. plantarum 299v
gives rise to a significant increase in the fecal concentra-
tion of short-chain fatty acids (from 83 to 113 mmol g–1
wet feces), as well as of acetic and propionic acids.
116 Lactic Acid Bacteria | Lactobacillus spp.: Lactobacillus plantarum
Moreover, a significant decrease in Gram-negative
anaerobic bacterial count has been observed in jejunum
and rectum of healthy volunteers after daily intake of
Lb. plantarum 299v and Lb. plantarum E98. Studies on the
effectiveness of Lb. plantarum against intestinal infections
and pathogenic bacteria also indicated a protective effect
against Escherichia coli and Clostridium difficile, and more
than one possible mechanism of action has been sup-
posed to explain this effect. Lactobacillus plantarum 299v
also showed an ability to reduce LDL-cholesterol and
fibrinogen levels in the blood. Administration of another
strain, Lb. plantarum PH04, reduced serum cholesterol by
7% and triglycerides by 10%, decreasing considerably
coronary heart diseases in rats. Moreover, a possible
mechanism of stimulation of the immune response in
mice has been reported after oral administration of
Lb. plantarum CRL 936.
Resistance to antibiotic action has been studied for
many Lb. plantarum strains isolated from dairy products.
Several strains of Lb. plantarum isolated from raw milk
showed a broad spectrum of resistance to many antibio-
tics such as tetracycline, erythromycin, ampicillin,
penicillin G, ofloxacin, and vancomycin. A certain sus-
ceptibility has been shown by some strains to
norfloxacin, gentamicin, amikacin, cefuroxime, and
streptomycin. Lactobacillus plantarum 423, tested as pro-
biotic in infant milk formulations, showed resistance to
norfloxacin and ciprofloxacin. Moreover, a high suscept-
ibility of Lb. plantarum to inhibitors of protein synthesis
has been demonstrated.
Many authors suggested that the use of Lb. plantarum as a
probiotic could be preferred above other lactic acid bac-
teria because of convenience in production, high-level
genetic accessibility, and high performance in the GI tract.
Other Characteristics of Lactobacillus
plantarum Possibly Related to Health Benefits
In the recent years, some work has been done to evaluate
the possibility of reducing the toxicity of some proteins
such as gluten by selected lactic acid bacteria: indeed,
the presence of residual proline and glycine polypep-
tides is considered to be responsible for celiac disease. A
probiotic named VSL#3, obtained with mixed cultures
of Lb. plantarum, other Lactobacillus spp., as well as
Streptococcus thermophilus and bifidobacteria, was success-
fully applied for the hydrolysis of gliadin during dough
leavening, and the resulting bread was tolerated by a
patient suffering from celiac disease. With relation to
the same food, as phytic acid and myo-inositol hexaphos-
phate (IP6) represent antinutritional factors in baked
products because of their property to chelate divalent
dietary minerals, a mixed starter culture including
Lb. plantarum, Lactobacillus brevis, and Lactobacillus curvatus
was successfully used to reduce IP6 content by about
80–90% in sourdough after 12 h of fermentation.
Recently, conjugated linoleic acids (CLAs), especially
the isomers cis-9, trans-11-octadecadienoic acid and trans-
10, cis-12-octadecadienoic acid, have attracted much
attention because of their beneficial effects, such as reduc-
tion of carcinogenesis, arteriosclerosis, and body fat. In
that context, it has been demonstrated that Lb. plantarum is
able to produce considerable amounts of cis-9, trans-11-
octadecadienoic acid and trans-9, trans-11-octadecadienoic
acid, the two main bioactive CLA isomers, by the
bioconversion of linoleic and ricinoleic acids.
Another challenge is represented by the use of
Lb. plantarum as a live vaccine to deliver heterologous
antigens to the mucosal immune system. For this pur-
pose, Lb. plantarum NCIMB 8826 has been studied as a
model; some studies have also exploited the ability of
Lb. plantarum to secrete bioactive molecules that evoke
an immune response. Advantages of the LAB over
traditional vaccines are represented by the noninvasive-
ness and the possibility to induce both a systemic and
mucosal immune response.
Milk and Dairy Products
Lactobacilli are microorganisms traditionally regarded as
useful for dairy technology, and interest in their applica-
tions is reinforced by documented probiotic and functional
properties. Lactobacillus plantarum has been frequently iso-
lated from dairy products, and it is a member of a group of
mesophilic lactobacilli referred to as ‘nonstarter lactic acid
bacteria’ (NSLAB), which represent an adventitious sec-
ondary microflora that can survive pasteurization and
enter milk or curd as a contaminant and become dominant,
reaching levels of 106–108 cfu g–1, during cheese ripening.
It has been reported that the count of Lb. plantarum is
higher during the first stage of cheese ripening (from
1 week to 3–4 months), reaching often more than
107 cfu g–1; during prolonged aging (6–7 months), the
count of the species generally decreases to 104 cfu g–1,
and often other Lactobacillus spp. become the dominant
microbial population. The growth rate and final popula-
tion density are affected by pH, salt, and moisture levels of
the curd and by the temperature of ripening. The presence
of Lb. plantarum has been documented in many cheese
varieties made throughout the world, using either pasteur-
ized or raw cows’, ewes’, or goats’ milk, as well as in
traditional fermented milks and milk-based beverages.
Some examples of European cheeses include Italian vari-
eties of Pecorino cheese and Gorgonzola, the Spanish
Manchego and Roncal, the Portuguese Picant, and Irish
Cheddar cheese. Among cheeses produced worldwide and
fermented milks, one can mention Tibetan Qula cheese,
Kenian fermented milk, Himalayan ethnic fermented
 Lactic Acid Bacteria | Lactobacillus spp.: Lactobacillus plantarum
milk, Moroccan camel milk, and New Zealand Cheddar
cheese.
In Feta and Teleme cheeses (Greece), Lb. plantarum
represents 47.8 and 65.8% of the isolated lactobacilli,
respectively; it constitutes 56.9% of the main species
isolated from Tenerife goats’ milk cheese, and it is pre-
dominant in cave and pit (Fossa) ripened cheeses. A study
on Irish Cheddar cheese indicated that Lb. plantarum
represented 28% of the NSLAB, while a similar study
on mature cheese revealed Lb. plantarum as 2.1% of the
population. Another study on the population dynamics of
Cheddar cheese referred to a mixture of Lb. plantarum,
Lb. paracasei, and Lb. rhamnosus till 6 weeks of maturation;
thereafter, no strains of Lb. plantarum were found.
Many laboratory-scale studies have been carried out in
model cheese systems to elucidate the dynamics and
impact of an ‘NSLAB starter’ formulation. Model studies
applied to Cheddar cheese revealed that NSLAB produce
desirable flavor and reduce harshness and bitterness asso-
ciated with some starter cultures; moreover, an extension
of shelf life of the cheeses was observed as well as a major
agreement at sensory analysis with respect to cheeses
made with traditional starter cultures.
Because the starter microorganisms rapidly deplete all
the lactose, the ability to use residual substrates such as
lactate, citrate, pyruvate, proteins, and lipids is necessary
for NSLAB. Various studies reported the ability of
Lb. plantarum to metabolize citrate in the presence or absence
of other fermentable sugar; citrate represents the precursor
of important flavor components of cheeses such as diacetyl,
acetate, acetoin, and 2,3-butanediol. Recent studies evaluat-
ing the interactions between starter bacteria and adjunct Lb.
plantarum showed that citrate degradation in cheese varied
according to the type of starter used; Lb. plantarum degraded
citrate to acetoin and diacetyl, especially in the presence of a
Cit– mesophilic homofermentative starter. Moreover, the
same adjunct strain degraded citrate to succinic acid in the
presence of a thermophilic starter.
In general, NSLAB show intense secondary proteolysis,
as evidenced by high oligoendopeptidase and aminopepti-
dase activities, leading to high levels of free amino
acids contributing directly or as precursors to cheese flavor.
Lactobacillus plantarum can catabolize aromatic and branched-
chain amino acids via transamination, which represents a
key step in the amino acid conversion to aroma compounds
in cheeses. The mechanism is initiated by a transamination
reaction and is catalyzed by aminotransferases, because the
degradation occurs only in the presence of an -ketoacid,
which is used as an amino group acceptor; flavor compounds
such as hydroxyacids, aldehydes, alcohols, and carboxylic
acids are further produced from -ketoacids.
Some studies have been focused on the purification
and characterization of lipases and esterases from
Lb. plantarum, indicating the potential significance of
some strains to lipolysis during cheese ripening and
117
highlighting the need for selecting appropriate starters
to produce enzyme-modified cheese as well as acceler-
ated ripened cheese because of wide variations in activity
between strains. It has been reported that strains of
Lb. plantarum isolated from Argentinean goats’ milk and
cheeses also showed high specific activity on -naphthol
butyrate, caproate, and acetate, and it was found that
some strains presented more than one esterase; for exam-
ple, Lb. plantarum O236 showed four enzymes that
hydrolyze carboxyl ester linkages with different specifi-
cities. Moreover, the intracellular and extracellular
fractions of the above strain were also able to hydrolyze
tributyrin. On the other hand, another strain (Lb. plantarum
O155) did not hydrolyze triglycerides.
Mesophilic lactobacilli also possess some glycoside
hydrolase activity, and it has been shown in model sys-
tems that they can utilize sugars from glycoproteins of the
milk fat globule membrane.
Even if the role of NSLAB in cheese ripening con-
tinues to be under investigation, the above-mentioned
finding corroborates the hypothesis of a useful impact of
selected Lb. plantarum strains on the overall quality of
many dairy products.
See also: Bacteria, Beneficial: Bifidobacterium spp.:
Applications in Fermented Milks. Fermented Milks:
Asian Fermented Milks; Buttermilk; Health Effects of
Fermented Milks; Kefir; Koumiss; Middle Eastern
Fermented Milks; Nordic Fermented Milks; Starter
Cultures; Types and Standards of Identity; Yogurt: Role
of Starter Culture; Yogurt: Types and Manufacture.
Nutrients, Digestion and Absorption: Fermentation in
the Rumen.
Further Reading
Axelsson LT (1993) Lactic acid bacteria: Classification and physiology.
In: Salminen S and von Wright A (eds.) Lactic Acid Bacteria,
pp. 1–63. New York: Marcel Dekker.
Beresford TP, Fitzsimons NA, Brennan NL, and Cogan TM (2001)
Recent advances in cheese microbiology. International Dairy Journal
11: 259–274.
Boekhorst J, Wels M, Kleerebezem M, and Siezen RJ (2006) The
predicted secretome of Lactobacillus plantarum WCFS1 sheds
light on interactions with its environment. Microbiology
152: 3175–3183.
De Vries MC, Vaughan EE, Kleerebezen M, and de Vos WM (2006)
Lactobacillus plantarum: Survival, functional and potential probiotic
properties in the human intestinal tract. International Dairy Journal
16: 1018–1028.
Ferain T, Schanck AN, and Delcour J (1996) 13C nuclear magnetic
resonance analysis of glucose and citrate end products in a ldhL-
ldhD double-knockout strain of Lactobacillus plantarum. Journal of
Bacteriology 178: 7311–7315.
Fimland N, Rogne P, Fimland G, Nissen-Meyer J, and Kristiansen PE
(2008) Three-dimensional structure of the two peptides that
constitute the two-peptide bacteriocin plantaricin EF. Biochimica et
Biophysica Acta 1784: 1711–1719.
118 Lactic Acid Bacteria | Lactobacillus spp.: Lactobacillus plantarum

Fox PF, McSweeney PLH, Cogan TM, and Guinee TP (eds.) (2004)
Cheese: Chemistry, Physics and Microbiology, Vol. 1: General Aspects,
3rd edn. Amsterdam, The, Netherlands: Elsevier Academic Press.
Hammes WP and Vogel RF (1995) The genus Lactobacillus.
In: Wood BJB and Holzapfel WH (eds.) The Genera of Lactic Acid
Bacteria, pp. 19–54. London: Blackie Academic and Professional.
Kleerebezem M, Boekhorst J, van Kranenburg R, et al. (2003) Complete
genome sequence of Lactobacillus plantarum WCFS1. Proceedings
of the National Academy of Sciences of the United States of America
100(4): 1990–1995.
Molenaar D, Bringel F, Schuren FH, de Vos WM, Siezen RJ, and
Kleerebezem M (2005) Exploring Lactobacillus plantarum genome
diversity by using microarrays. Journal of Bacteriology
187(17): 6119–6127.
Pfeiler EA and Klaenhammer TR (2007) The genomics of lactic acid
bacteria. Trends in Microbiology 15: 546–553.
Skeie S, Kieronczyk A, Eidet S, Reitan M, Olsen K, and Østlie H (2008)
Interaction between starter bacteria and adjunct Lactobacillus
plantarum INF15D on the degradation of citrate, asparagine and
aspartate in a washed-curd cheese. International Dairy Journal
18: 169–177.
Yvon M and Rijnen L (2001) Cheese flavour formation by amino acid
catabolism. International Dairy Journal 11: 185–201.
Relevant Websites
http://www.lacplantcyc.nl – LacplantCyc: A Lactobacillus
plantarum Pathway/Genome Database as a Reference for
Lactic Acid Bacteria.