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Agricultural Research Council, Research Centre for Forage and Dairy Productions (CRA-FLC), Via Lombardo 11, 26900 Lodi, Italy
b
The G. Eliava Institute of Bacteriophage, Microbiology and Virology, 0160 Tbilisi, Georgia
c
Research Center for Biotechnology, Indonesian Institute of Sciences, 16911 Cibinong, Indonesia
Received 14 October 2009; accepted 2 March 2010
Available online 17 March 2010
Abstract
The genus Lactobacillus is a heterogeneous group of lactic acid bacteria (LAB) with important implications in food fermentation. The ability
to colonize a variety of habitats is a direct consequence of the wide metabolic versatility of this group of LAB. Consequently, lactobacilli have
been used for decades in food preservation, as starters for dairy products, fermented vegetables, fish and sausages as well as silage inoculants.
Lactobacilli have also been proposed as probiotics and microbial cell factories for the production of nutraceuticals. However, a wide range of
applications of lactobacilli in food biotechnology remains potential, whereas a number of important strains still need to be discovered and
characterized. This article provides an overview of the taxonomy of lactobacilli and describes four of the most significant case studies on the
application of this group of LAB in food and feed biotechnology, including their use as probiotics, dairy starters, silage inoculants, and microbial
cell factories. The importance of access to and exchange of biological material within and between different strain collections as a crucial step in
expanding the range of different biotechnological applications of lactobacilli is also emphasized.
2010 Elsevier Masson SAS. All rights reserved.
Keywords: Lactobacillus spp.; Probiotic lactobacilli; Dairy starter cultures; Silage inoculants; Microbial cell factories
1. Introduction
The genus Lactobacillus belongs to the large group of lactic
acid bacteria (LAB) which are all Gram-positive organisms
which produce lactic acid by fermentation. Genera of LAB
include, among others, Lactococcus, Enterococcus, Oenococcus, Pediococcus, Streptococcus, Leuconostoc and Lactobacillus (Kandler and Weiss, 1986). With over 100 species and
subspecies, the genus Lactobacillus represents the largest group
within the family Lactobacillaceae. Members of the genus are
rod-shaped, often organized in chains. They are strictly
fermentative and aerotolerant, but grow well under anaerobic
conditions. There are two groups of species depending on the
ability to ferment sugars: homofermentative species, converting
sugars mostly into lactic acid, and heterofermentative species,
converting sugars into lactic acid, acetic acid, ethanol and CO2.
Because the main catabolite is lactic acid, lactobacilli prefer
relatively acidic conditions (pH 5.5e6.5).
Bacteria belonging to the genus Lactobacillus (lactobacilli)
can be found in a variety of ecological niches such as plants,
animals and raw milk (Hammes and Vogel, 1995). In addition,
lactobacilli can be found in insects. The ability to colonize
such a variety of habitats is a direct consequence of the wide
metabolic versatility of this group of LAB. Hence, it is not
unexpected that lactobacilli have been used for decades in
food preservation, as starters for dairy products, fermented
vegetables, fish and sausages as well as silage inoculants.
Because of their potential therapeutic and prophylactic attributes, lactobacilli have also been proposed as probiotics.
1.1. Taxonomy and phylogeny
* Corresponding author.
E-mail addresses: giorgio.giraffa@entecra.it (G. Giraffa), n_chanish.
ibmv@caucasus.net (N. Chanishvili), yantyatiwidyastuti@yahoo.com (Y.
Widyastuti).
0923-2508/$ - see front matter 2010 Elsevier Masson SAS. All rights reserved.
doi:10.1016/j.resmic.2010.03.001
16S/23S rRNA gene sequences revealed phylogenetic relationships among lactobacilli. Collins et al. (1991) originally
distinguished three phylogenetic groups, the Lactobacillus
caseiePediococcus, Lactobacillus delbrueckii, and Leuconostoc group. Later, the L. delbrueckii group was renamed as
the Lactobacillus acidophilus group (Schleifer and Ludwig,
1995) and the L. caseiePediococcus group was split into the
L. casei group, the Lactobacillus plantarum group, the
Lactobacillus reuteri group, the Lactobacillus buchneri group
and the Lactobacillus salivarius group. At present, the
phylogenetic structure of the genus Lactobacillus also
includes the Lactobacillus perolens group, the Pediococcus
group, and the Lactobacillus vitulinusecatenaformis group
(Pot and Tsakalidou, 2009).
The L. acidophilus group contains almost exclusively
obligately homofermentative lactobacilli. Besides L. acidophilus, many other species of industrial interest such as
Lactobacillus amylovorus, Lactobacillus crispatus, L. delbrueckii with the subspecies delbrueckii, lactis, bulgaricus,
and indicus, Lactobacillus johnsonii, Lactobacillus helveticus,
and Lactobacillus gasseri are included. L. helveticus and L.
delbrueckii play an acknowledged role as starters in dairy and
vegetable fermentation, respectively, whereas some strains of
the species L. acidophilus have long been known to play a key
role in human health and nutrition by their positive influence
on the intestinal flora (Pot, 2008).
The L. casei group contains the most well-known species
Lactobacillus rhamnosus, Lactobacillus paracasei, L. casei, and
Lactobacillus zeae. This subgroup has been subjected to extensive taxonomic revision in the past, leading to a provisional
rejection of the species name L. paracasei. At the moment, these
species are still considered separate species within the genus
(Dellaglio et al., 2002; Desai et al., 2006; De Vos et al., 2005;
Pot, 2008). L. paracasei and L. rhamnosus are the most
common foodborne isolates within the L. casei group of lactobacilli, especially from cheese. L. paracasei and L. casei are also
found in silage and are common inhabitants of the animal/human
gastrointestinal tract (Pot and Tsakalidou, 2009).
The L. plantarum group encompasses the taxa L. plantarum
subsp. plantarum, L. plantarum subsp. argentoratensis, Lactobacillus paraplantarum, and Lactobacillus pentosus. L. plantarum is widely found in a range of food such as dairy, meat, and
vegetable products. It is commonly found in the human
gastrointestinal tract as a consequence of its proven ability to
survive gastric transit and colonize the gut. L. plantarum is
considered a food-grade microorganism because of its long and
documented history of safe use in fermented foods (de Vries
et al., 2006).
The L. reuteri group contains at least six species exclusively
isolated from sourdough. It should be noted that L. reuteri
strains may also be dominant members of sourdough populations (Del Bello et al., 2005). Lactobacillus fermentum, one
of the best-known species of this group, has been isolated from
vegetable and dairy fermentation (Pot, 2008; Pot and
Tsakalidou, 2009).
The L. buchneri group is a large and heterogeneous group
of lactobacilli mostly linked to food fermentations. To this
481
482
Table 1
The most commonly used species of lactobacilli in probiotic preparations
(especially in dairy products) (Keohane et al., 2009; Ouwehand et al., 2002;
Parvez et al., 2006; Rastall et al., 2005).
Species
Main commercially
used strain(s) (when available)
with documented health benefits
Lactobacillus
Lactobacillus
Lactobacillus
Lactobacillus
acidophilus
johnsonii
(para)casei
rhamnosus
Lactobacillus
Lactobacillus
bulgaricus
Lactobacillus
lactis
Lactobacillus
Lactobacillus
Lactobacillus
Lactobacillus
Lactobacillus
Lactobacillus
Lactobacillus
Lactobacillus
Lactobacillus
Lactobacillus
Lactobacillus
plantarum
delbrueckii subsp.
LA1; LA5
La1; NCFM; DDS-1; SBT-2062
F19; CRL 431; Immunitass; Shirota
GG; LB21; 271; GR-1;
VTT E-97800
299v; Lp01
Lb12
delbrueckii subsp.
Lla
cellobiosus
curvatus
fermentum
reuteri
brevis
salivarius
helveticus
amylovorus
crispatus
gallinarum
gasseri
RC-14
MM2
UCC118
B02
LG21
483
484
Table 2
Lactic acid bacteria in silage.
Species
Forage
Reference
Lactobacillus plantarum
Lactococcus lactis
Leuconostoc
pseudomesenteroides
Pediococcus acidilactici
Lactobacillus brevis
Enterococcus faecalis
Weissella kimchii
Pediococcus pentosaceus
Lactobacillus plantarum
Pediococcus pentosaceus
Lactobacillus homohiochii
Lactobacillus brevis
Lactobacillus gasseri
Lactobacillus plantarum
Lactobacillus coryniformis
Leuconostoc spp.
Enterococcus faecium
Enterococcus faecalis
Pediococcus acidilactici
Pediococcus pentosaceus
Pediococcus spp.
Lactobacillus brevis
Lactobacillus fermentum
Leuconostoc mesenteroides
Paddy rice
(Oryza sativa)
Ennahar
et al., 2003
Alfalfa
(Medicago sativa L.)
Lin
et al., 1992
Pangola (Digitaria
decumbens), Setaria
(Setaria sphacelata) and
Hamil (Panicum maximum)
Tjandraatmadja
et al., 1994a,b
Fig. 1. Silage preparation using lactobacilli as inoculants. A: chopping grass; B: adding rice bran and inoculants as additives; C: silage inoculants; D: silage
fermentation in plastic bags.
485
486
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