O A

999
Advances in Environmental Biology, 7(6): 999-1007, 2013
ISSN 1995-0756
This is a refereed journal and all articles are professionally screened and reviewed
ORIGINAL ARTICLE
Diversity And Technological Properties Of Predominant Lactic Acid Bacteria Isolated
From Algerian Raw Goat’s Milk
1
Moulay M., 3Benlahcen K. 2Aggad H. and 3Kihal M.
1
Department of Natural and Life Sciences, Faculty of Natural and Life Sciences.Ibn Khaldoun University Tiaret.
Algéria
2
Faculty of Sciences agro-vétérinairy, University Ibn Khaldoun, BP 78 , Tiaret. Algeria
3
Laboratory of Applied Microbiology, University of Oran. Department of Biology, Faculty of Sciences, Oran
University BP 16. Es-Sénia, 31100, Oran, Algeria.
Moulay M., Benlahcen K. Aggad H. and Kihal M.: Diversity And Technological Properties Of
Predominant Lactic Acid Bacteria Isolated From Algerian Raw Goat’s Milk
ABSTRACT
Lactic acid bacteria (LAB) are a group of gram-positive, lactic acid producing firmicutes. They have been
extensively used in food fermentations, including the production of various dairy products. The proteolytic
system of LAB converts proteins to peptides and then to amino acids, which is essential for bacterial growth and
also contributes significantly to flavor compounds as end-products. The objective of this paper is the
microbiological and technological characterization of lactic acid bacteria isolated from Algerian raw goat’s
milk. Microbiological, physiological and biochemical tests were made for the work performed. Dominant lactic
acid bacteria were isolated on M17 and MRS media from Algerian goat's milks, either raw and/or fermented
under laboratory conditions. Isolates were characterized for three phenotypes essential for Raib manufacture: (a)
high acidifying activity over the normal associated temperature range, (b) presence of protease, and (c) ability to
metabolize citrate. Thirty nine isolates were characterized and 13 strains which have been found to belonging to
lactic acid bacteria including the species of (Lactococcus lactis subsp. lactis and Lactococcus lactis subsp.
cremoris), Leuconostoc (5 isolates) and Lactobacillus (6 isolates). These strains lactic have technological
characteristics acidification and interesting flavor. These characters of industrial interest can be exploited to
provide a specific starter for the local dairy industry.
Key words: Goat, raw milk, lactic acid bacteria, Isolation, Identification, Acidity.
Introduction
Lactic acid bacteria (LAB) have a very long
history of use in the manufacturing processes of
fermented foods and a great deal of effort was made
to investigate and manipulate the role of LAB in
these processes. Lactic acid bacteria (LAB) form a
phylogenetically diverse group of gram-positive
bacteria used worldwide in various traditional and
industrial food fermentations, including probiotic
products. As such, Lactococcus lactis, Streptococcus
thermophilus,
several
Lactobacillus
and
Bifodobacterium species, have a significant history
of safe use, especially in dairy products.
Today, the diverse group of LAB includes
species that are among the best-studied
microorganisms and proteolysis is one of the
particular physiological traits of LAB of which
detailed knowledge was obtained [25]. The
proteolytic system involved in casein utilization
provides cells with essential amino acids during
growth in milk and is also of industrial importance
due to its contribution to the development of the
organoleptic properties of fermented milk products
[34,13,33].
In Algeria the goat population has increased in
recent decades, which implies growth in milk
production in arid and mountainous [2,26]. Goat's
milk undergoes spontaneous fermentation and is
often consumed as traditional dairy products (Lben,
Raib and Jben) [3,5]. Characterization of metabolic
activities of new strains of lactic acid bacteria
isolated from raw goat's milk and dairy processing
operation in the production of organic-functional
ingredients is one of the axes of universal search
[12].
New sources of nutrients should be more
exploited for varying the human diet and also to
benefit from new functional ingredients and natural
food components. African and Arab countries, where
the breeding conditions for cows are severe and
fastidious, can get over this situation by developing a
Corresponding Author
Moulay Meriem, Department of Natural and Life Sciences, Faculty of Natural and Life
Sciences.Ibn Khaldoun University Tiaret. Algéria
E-mail: [email protected]
1000
Adv. Environ. Biol., 7(6): 999-1007, 2013
breeding system for local animals such as gaots. The
beneficial microbiota of gaots milk represented by
LAB is a potential source of biological materials to
be used in dairy technology. The transformation of
gaots milk by fermentation is not easy and more
research for elucidating the process is needed
[43,44]. Goat's milk is traditionally fermented by
lactic microflora whose species composition is not
stable. Transforming traditional goat's milk is made
by spontaneous fermentation by lactic microflora
undefined, which often produces different sensory
quality [24]. The progresses of the dairy industry
have allowed manufacturers to build mixture of lactic
strains with precise proportions which allowed the
production to standardize various types of fermented
milks and cheeses.
If cow milk was widely investigated, up until
now, little studies were undertaken on the Algerian
gaots milk to characterize its micro
flora especially
LAB. The aim of the present paper is to determine
the biochemical, physiological and phenotypic
properties of new strains of LAB isolated from the
raw gaot’s milk produced west Algeria.
Materials And Methods
Samples collection:
Samples of goat’s milk were were collected
from individual households in rural areas of different
places in west Algeria (Es-senia, Oran and Tiaret),
Samples were collected aseptically in sterile bottles
(500 mL), leveled kept in an ice-box container (4°C)
and transported to the laboratory for microbiological
analyses [3].
colonies will perform Gram stain and catalase search.
Isolates gram(+) and catalase(-) are retained. Colonies
of pure isolates were inoculated on liquid culture
medium: The medium M17 and MRS medium (pH =
6.7) were used for the growth of cocci, bacilli for the
MRS medium (pH = 5.4) was chosen. After 24 h of
incubation at 30 ° C the aspect of culture was noted
[34,43].
Conservation strains :
The strains of lactic acid bacteria were stored
without appreciable loss of properties in skimmed
milk with 30% (v/v) glycerol at -20°C [2,8].
Working cultures were also kept on MRS agar or
M17 agar slant at 4° C and streaked every 4 weeks
[22,4,43].
Characterization and identification of strains:
Macroscopic examination to describe the
bacterial colonies on solid medium: It is the color,
edge, elevation, aspect, pigmentation, opacity and
diameter of colonies. The appearance of the culture
liquid medium is also observed. This characterization
is carried out on pure isolates on which test
biochemical
reactions
necessary
for
their
differentiation. Tests were carried out on each
isolate. These were the following: Gram and
morphology, catalase, gas production from glucose,
hydrolysis of arginine, growth in 4 and 6.5% NaCl
broth, dextran production, growth at pH 9.6 and in
40% bile (cocci) and growth at 10 and 45 °C.
Microscopic examination defined cell morphological
appearance such as shape, pairing mode and type
isolates of gram [17,11].
Isolation of lactic microflora:
Test milk Sherman:
Samples (10 g) of each samples of raw milk
were mixed with 90 ml of 0.85% (w/v) sterile
physiological saline and homogenized in a vortex for
1 min. A serial dilution in the same diluents was
made. The last 3 dilutions (10-7, 10-8, 10-9) are seeded
deep culture media MRS and M17, following the
method of Guiraud [18]. Lactic acid bacteria (LAB)
were enumerated on MRS agar and incubated at
30°C for 48–72 h. Representative LAB strains were
isolated from MRS plates of the highest sample
dilutions. Colonies were selected randomly or all
sampled if the plates contained less than 10 colonies.
Purity of the isolates was checked by streaking again
and subculturing on fresh MRS broth as well as MRS
agar, followed by microscopic examinations.
Purification of isolates:
After incubation, colonies with different
morphological aspects (4 to 6 colonies) were purified
by streaking on MRS or M17 agar, then incubating
the tubes at 30 °C for 24 hours. Obtaining pure
Bacterial strains were seeded in sterile skim milk
at 0.1% and 0.3% methylene blue; the whole is
incubated at 30°C for 24h to 48h. A positive result is
defined as the coagulation of milk and reduction of
the dye. This test is particularly useful for
differentiating faecal streptococci and lactococci.
Lactococci are capable of growing in the presence of
0.1% methylene blue, enterococci grow in the
presence of 0.3% [37].
Test citrate:
The test is performed using citrate agar medium
of Kempler and McKay [20], containing ferric citrate
and potassium ferrocyanide. Strains tested, and after
incubation at 30 ° C for 24h to 48h, those using
citrate give blue colonies.
Arginine dihydrolase (ADH):
1001
Adv. Environ. Biol., 7(6): 999-1007, 2013
Inoculating strains on M16BCP medium [38]
and incubated at 30 °C for 24 h to 48 h. Culture
manifests itself by a transfer of the color purple to
yellow due to environmental glucose metabolism,
degradation of arginine and ammonia release
prevents the transfer of yellow. Cultures with
arginine dihydrolase will alkalize the middle.
Dextran test:
Dextran production is performed on MSE agar
medium rich in sucrose. Dextran producing strains
give viscous colonies [32].
Acétoїne production:
Clark and Lubs medium was used, after
inoculating strains in 10 ml of this medium, the
cultures were incubated at 30 °C for 24h. Then the
reaction of Vosges-Proskaur (VP) in 0.5 ml of
reagent adding α-naphthol to 6% in absolute alcohol
(VPI) and 0.5 ml of sodium hydroxide (NaOH) to
16% in the distilled water (VPII). The tubes are then
stirred, after 10 min of rest the presence of a pink
ring on the surface of the culture shows the positive
response [19].
Utilization of carbohydrates:
Fermentation of carbohydrates in traditional
galleries tubes of liquid medium MRS and M17
containing bromocresol purple (0.04 g/l) as a pH
indicator and supplemented with 1% carbohydrate.
Carbohydrates tested were: glucose, galactose,
fructose, mannose, lactose, xylose, raffinose,
arabinose, mannitol and sucrose. The positive result
is expressed as yellowing of the medium which is
due to the degradation of sugar which acidifies the
medium [3].
Kinetics of lactic acid production:
Precultures were prepared in skim milk medium
supplemented with 0.1% yeast extract and incubated
at 30 °C until coagulation. 100 ml sterile skim milk
was inoculated with 0.1% of the preculture. After
gentle agitation culture is divided into tube
(10ml/tube) and incubated at 30 °C. At a regular
interval time, samples were aseptically collected
every 2 h. A volume of 1 ml culture samples were
submitted to decimal dilutions in sterile saline
solution. Then 0.1 ml of the appropriate dilution was
inoculated in MRS agar medium to assess the
number of cells Mathot et al., [30]. The plates were
incubated at 30 °C for 48 h. Only plates containing
between 25 and 250 colonies were retained. The
generation time and growth rate were calculated in
the exponential growth phase [21]. The kinetics of
the changes in pH and acidity were also followed by
measuring pH and dornic acidity [34,21].
Results And Discussion
From 138 isolates obtained from raw goat's milk,
we have selected 13 representative strains that were
lactic acid bacteria. The macroscopic morphological
study on both media M17 and MRS agar colonies
revealed small lenticular, sometimes circular, about
1mm in diameter, whitish or slightly yellowish,
smooth edges and regular (Tab 1). Colonies on MRS
medium are smaller. Microscopic observation
showed that the isolates obtained on M17 medium
are gram-positive cocci and catalase negative. In
contrast to colonies on MRS medium size appeared
different. Growth on M17 medium or MRS liquid
produces a disorder in the bottom of the tube surface
area remains clear because the lactic acid bacteria
grow best at a low pressure of oxygen. Microscopic
observation showed the presence of two types of cell
morphology, the small colonies give a rode cells and
large colonies give cocci cells. The isolates were
separated into two groups, the first group of cocci
cells (SH1, SH2, SH3, SH4, SH4, SH5, SH6 and
SH7) and the second group of rode cells (SH8, SH9,
SH10, SH11, SH12 and SH13).The results presented
in Table (01) illustrate some specific characteristics
of the thirteen isolates. Goat's milk has different
types of lactic acid bacteria. Work of Tserovsk et al.,
[39] and Kostinek [23], confirm that the species of
lactic acid bacteria isolated from raw goat's milk is in
the form cocci and bacilli is diploїdes or associated
in clusters or chains and are Gram-positive, catalase-,
micro-aerophilic, acidtolerant, non-sporulating and
strictly fermentative with lactic acid as the major end
product during sugar fermentation.
The group of cocci (SH1, SH2, SH3, SH4, SH4,
SH5, SH6 and SH7) was selected due to their
inability to grow at 63.5 °C (thermal resistance), pH
9.6 (alkaline-broth), 6.5% NaCl (hypersaline broth)
and 0.3% methylene blue (Tab. 2). Isolates that grow
at this temperature are considered enterococci and
subsequently were excluded. The work of Badis et
al. [4] found enterococci isolating lactic acid bacteria
from raw goat's milk. Reduction of methylene blue in
0.3% skim milk has allowed the confirmation of
membership enterococci. Only isolates gram (+) cocci
and catalase (-) do not grow to 0.3% methylene blue
milk are retained. The second group sticks were
seeded on acidified MRS medium and incubated at
45 °C, 30 °C and 15 °C for 24 hours. The results
obtained show that the microscopic cells are nonspore forming rods Gram positive and catalase
negative. All isolates grown at 30 °C but some have
the ability to grow well at 15 °C and others at 45 °C.
According to studies Badis et al., [4], six isolates
belong to the genus Lactobacillus, which can be
mesophilic (SH10, SH12, SH13) or thermophiles
(SH8, SH9, SH11).
Growth on PCA medium with milk allows to
detecte the proteolytic strains which can produce a
1002
Adv. Environ. Biol., 7(6): 999-1007, 2013
clear halo near the colony. All isolates were
proteolytic. The use of citric acid was tested on
KMK medium, blue colonies were formed when the
citratase was produced. All rods form (SH8, SH9,
SH10, SH11, SH12 and SH13) can use citrate and
produce blue colonies. In contrast, only (SH1, SH3,
SH4 and SH6) (cocci) were able to produce blue
colonies on medium Kempler and McKay. Type
fermentative cocci isolates revealed that 05 isolates
were heterofementatives (SH1, SH3, SH4, SH5 and
SH6) and tow isolates (SH2 and SH7) were
homofermentatives. In heterofermentative groups, 04
isolates (SH1, SH4, SH5 and SH6) produce dextran
and
give
viscous
colonies
(Tab.2).
Table 1: Characterization microscopic of strains isolated
Strains
Catalase
Gram
Aspect of the colony
SH1
lenticular cream
+
SH2
lenticular cream
+
lenticular cream
SH3
+
lenticular cream
SH4
+
lenticular cream
SH5
+
lenticular cream
SH6
+
lenticular cream
SH7
+
small colony whitish
SH8
+
small colony whitish
SH9
+
small colony whitish
SH10
+
small colony whitish
SH11
+
small colony whitish
SH12
+
small colony whitish
SH13
+
All rods form strain did not produce dextran and
only (SH10 and SH13) were homofermentative
(Tab.2). The isolate is the only SH2 (cocci) and two
rods isolates (SH9 and SH13) which can hydrolyze
arginine.The SH11 strain is able to acidify milk
quickly.
The results show that the raw goat's milk
contains a high variety of lactic acid bacteria. Species
identification of lactic acid bacteria requires other
research tests that allow the differentiation between
species of the genus found. The accomplishments of
profile fermentation of differents sugars can
determine the species. Identifying species within the
genera Enterococcus, Lactococcus, Leuconostoc,
Pediococcus, Streptococcus, and Lactobacillus by
classical phenotypic methods presents particular
difficulties for microbiologists [15].
The
morphological,
physiological
and
biochemical characters showed that the five isolates
(SH1, SH3, SH4, SH5, SH6) are heterofementatives
cocci belonging to Leuconostoc genus. They are
unable to hydrolyze arginine, grow at 30 °C and not
at 45 °C. Some strains of Leuconostoc and
Lactococcus can use citrate which was unstable in
both genus [22,31,36],
Our results are consistent with the work of
Nokuthula et al., [35], which states that Leuconostoc
mesenteroides strains are capable of producing
dextran hydrolysis and citrate.
The two subspecies mesenteroides and
dextranicum differ in their sugar fermentation profil
[39,14,43]. Strain SH1 and SH3 unlike strain
produced dextran and ferment xylose and raffinose.
These physiological characteristics were similar with
the classification of Carr et al. [10] and are identified
respectively as, Leuconostoc pseudomesenteroides,
Leuconostoc mesenteroides.
The other two homofermentative cocci (SH2 and
SH7) belongs to the species Lactococcus lactis, the
The form
cocci
cocci
cocci
cocci
cocci
cocci
cocci
bacillus
bacillus
bacillus
bacillus
bacillus
bacillus
The mode of association
diplo in chainette
diplo in chainette
diplo in chainette
diplo in chainette
in chainette
in chainette
diplo
in chainette
diplo, in clusters
in chainette
diplo in chainette
diplo in chainette
diplo, in clusters
profile of sugar fermentation allowed to identify two
sub-species Lactococcus lactis subsp. lactis (SH2)
and Lactococcus lactis subsp. cremoris (SH7). These
subspecies differ in their fermentation profile by their
ability to hydrolyze arginine [30,37,16].
The analysis of morphological, physiological
and biochemical characters of second group of rod
cellular form which is compared to the characteristics
of Lactobacillus species described by Atlan et al.,
[1], Carr et al., [10] Dworkin et al., [14], Liu et al.,
[26] and Mami et al., [27] confirmed the
identification of the six strains: SH8, SH9, SH10,
SH11, SH12, SH13, respectively belonging to the
following species: Lactobacillus lactis, Lactobacillus
rhamnosus, Lactobacillus plantarum, Lactobacillus
bulgaricus, Lactobacillus brevis, Lactobacillus sp.
Some authors Atlan et al. [1], Varmanen et al., [40]
and Varmanen et al., [41] have isolated proteolytic
species of Lactobacillus such as Lactobacillus
bulgaricus and Lactobacillus rhamnosus with
aminopeptidases and prolinase activity. This
diversity of species found in this study is relative and
dependent primarily on the nature of the material
isolated and the different criteria used for each study,
as reported by Bissonnette et al., [7].
Kinetics of acidification:
Power acidifying strains is made of milk
medium. Acidification of milk is mainly due to the
production of lactic acid [34]. The results show that
all strains acidified milk. It's probably the screening
of proteolytic strains. Growth strongly proteolytic
strains medium such as Lactococcus lactis subsp.
lactis SH2 and Lactococcus lactis subsp. cremoris
SH7 gave a generation time of 48.38 min and 90
min, respectively. The stationary phase was reached
after 14 hours of incubation and the maximum
observed cell reached 10 log. This difference in
1003
Adv. Environ. Biol., 7(6): 999-1007, 2013
growth rate is lower in Lactococcus lactis subsp.
cremoris SH7 (μ = 0.66 h-1) because this strain have
reduced fermentation profile of carbohydrate
compared to Lactococcus lactis subsp. lactis SH2 (μ
= 1.24 h-1) (Fig. 3).
Table 2: Phenotype characteristics of lactic acid bacteria isolated from Algerian raw goat’s milk and the profile fermentation of sugars.
Strains
SH
SH
SH
SH
SH
SH1
SH4
SH7
SH9
SH10
SH11
SH12
SH13
Characters
2
3
5
6
8
cell shape
Cocc Cocci Coc Cocci
Cocci Coc Cocci
Ro
Rod
Rod
Rod
Rod
Rod
i
ci
ci
d
Growth at
15 °C
Nt
Nt
Nt
Nt
Nt
Nt
Nt
+
+
+
+
30 °C
+
+
+
+
+
+
+
+
+
+
+
+
+
45 °C
+
+
+
63.5°C
Nt
Nt
Nt
Nt
Nt
Nt
NaCl 6.5 %
Nt
Nt
Nt
Nt
Nt
Nt
pH = 9.6
Nt
Nt
Nt
Nt
Nt
Nt
+
+
+
hydrolysis of arginine
+
+
+
+
+
+
+
+
+
+
Citrate
+
+
+
+
Dextrane
Het
Hom Het Het
Het Het Hom
Het Het
Hom
Het
Het
Hom
Type fermentation*
+
+
+
+
Acétoîne
+
+
+
+
+
+
+
+
+
+
+
+
+
Glucose
+
+
+
+
+
+
+
+
+
+
+
Galactose
+
+
+
+
+
+
Nt
Nt
Nt
Nt
Nt
Nt
Fructose
+
±
+
+
+
+
+
+
+
+
+
Mannose
+
+
+
+
+
+
+
+
+
+
+
+
+
Lactose
+
+
+
+
+
+
+
+
Xylose
+
+
+
+
+
+
+
Raffinose
+
+
+
+
Nt
Nt
Nt
Nt
Nt
Nt
Arabinose
+
+
+
+
+
Mannitol
+
±
+
+
+
+
Nt
Nt
Nt
Nt
Nt
Nt
Saccharose
Nt
Sorbitol
+
+
+
+
+
+
Nt
+
+
+
+
+
Maltose
Ribose
+
+
+
+
+
Nt
+
+
+
+
+
+
Milk Sherman
0,1% MB
+
+
+
+
+
+
+
+
Nt
Nt
Nt
Nt
Nt
0,3% MB
Nt
Nt
Nt
Nt
Nt
Hét = Hétérofermentation, Hom = Homofermentation, MB=Methylene Blue
SH1 : Leuconostoc pseudomesenteroïdes, SH2 : Lactococcus lactis subsp. lactis, SH3 : Leuconostoc paramesenteroïdes, SH4 :
Leuconostoc mesenteroïdes subsp. mesenteroïdes, SH5 : Leuconostoc mesenteroïdes subsp. mensenteroïdes. SH6 : Leuconostoc
mesenteroïdes subsp. dextranicum, SH7: Lactococcus lactis subsp. cremoris, SH8: Lactobacillus lactis, SH9 : Lactobacillus rhamnosus, SH10
: Lactobacillus plantarum, SH11 : Lactobacillus bulgaricus, SH12 : Lactobacillus brevis, SH13 : Lactobacillus sp.
The kinetics of lactic acid production is shown
in Figure 2. The amount of lactic acid produced after
26 h of incubation varied from 27 °D to 95 °D
among strains Leuconostoc mesenteroides subsp.
dextranicum (SH6) and Lactobacillus rhamnosus
(SH9) respectively. The amount of lactic acid
produced in function of time is slightly variable from
one strain to another. The species of Leuconostoc are
very important for fermented dairy products, as they
contribute to the organoleptic characteristics of butter
and cream, and also contribute to the formation of
openings in some soft, semi-hard (Edam and Gouda
cheeses), many artisanal or in blue-veined cheeses,
such as Roquefort [9]. Lactic acid bacteria have an
inefficient proteolytic system. Therefore, cultivation
media which may have high protein content are
usually supplemented with yeast extract or protein
lysates (peptones). These additives might be
conveniently replaced by in situ treatment of the
cultivation medium with proteolytic enzymes or
proteolytic microbes [42].
Species Lactococcus lactis subsp. lactis (SH2),
and Lactococcus lactis subsp. cremoris (SH7) have
the capacity to produce up to 100 °D and, 98 °D
lactic acid respectively (Fig.2). Work of [29,4], had
found the same results. There is a close relationship
between the pH and the Dornic acidity and the pH
decreases with increasing amount of lactic acid
produced. These results corroborate that our strains
have a performance acidification interesting for
integration in manufacturing milk. The technological
characterization of the isolates was performed in
order to evaluate the suitability of individuals to be
used as adjunct starter cultures in the manufacture of
fermented foods. The antimicrobial activity of
Leuconostoc mesenteroides isolated from Algerian
raw milk was accomplished by the work of
Benmechernene et al. [6].
1004
Adv. Environ. Biol., 7(6): 999-1007, 2013
7
6
5
4
pH
3
2
1
0
0
2
4
6
8
10
24
SH1
SH2
SH3
SH4
SH5
SH6
SH8
SH9
SH10
SH11
SH12
SH13
26
T (h)
SH7
Fig. 1 : Kinetics of changes in pH of the different isolates in milk medium.
120
°D
100
80
60
40
20
T (h)
0
0
2
4
6
8
10
24
SH1
SH2
SH3
SH4
SH5
SH6
SH8
SH9
SH10
SH11
SH12
SH13
26
SH7
Fig. 2 : Kinetics of acidification degree Dornic different isolates in milk medium.
Fig. 3: Kinetics of growth of Lactococcus lactis subsp. lactis SH2 (▲) and Lactococcus lactis subsp. cremoris
SH7 (■) in milk medium at 30 ° C.
Conclusion:
This study provides evidence that crude milk
was rich in biodiversity and can be considered as
adequate sources of new strains and species of lactic
acid bacteria. There are several factors which
influence the development of the organoleptic
characteristics of cheese, including the type of milk,
its microbiological quality, technology used in
making the cheese, the conditions of ripening, and
1005
Adv. Environ. Biol., 7(6): 999-1007, 2013
others. However, the lactic acid bacteria (LAB) play
a principal role in releasing speci
fic compounds
responsible
forflavour cheese
development.Technological characterization of acid
production and exo polysaccharide, slow autolysis
and
proteolytic
activity,
suggested
that,
fundamentally, the dominant lactic acid bacteria in
raw milk are not all interesting for industrial
application. This study identified isolates of lactic
acid bacteria from Algerian raw goat’s milk. The
study led to isolate 13 strains belonging to the genus
of Leuconostoc, Lactococcus and Lactobacillus.
These species are proteolytic and some of them are
fast acidifying milk. It seems that, to achieve this
goal, the use of these isolates as starter cultures and
assistants in local cheeses desirable. This study has
shown that adventitious lactic acid bacteria can be a
source of new strains with interesting technological
properties that could be used for fermented dairy
foods.
8.
9.
10.
11.
Références
1.
2.
3.
4.
5.
6.
7.
Atlan, D., C. Gilbert, B. Blanc and R. Portalier,
1994. Cloning, sequencing and characterization
of the pepIP gene encoding a proline
iminopeptidase from Lactobacillus delbrueckii
subsp. bulgaricus CNRZ 397. Microbiology,
140 (3): 527-535.
Badis, A., 2004. Identification et caractérisation
technologique de bactéries lactiques isolée à
partir de lait crue de chèvre de quatre
populations caprines locales. Thèse de doctorat
d’état, université d’Oran.
Badis, A., N. Laouabdia-Sellami, D. Guetarni,
M. Kihal and R. Ouzrout, 2005. Caractérisation
phenotypique des bactéries lactiques isolées à
partir de lait cru de chèvre de Deux populations
caprines locales Arrabia et Kabyle. Sci Technol.,
23: 30-37.
Badis, A., D. Guetarni, B. Moussa Boudjemaa,
D.J. Henni, M.E. Tornadijo and M. Kihal, 2004.
identification of cultivable lactic acid bacteria
isolated from Algerian raw goat’s milk and
evaluation of their technological properties.
Food Microbiol., 21: 343-349.
Bendimerad, N., M. Kihal, and F. Berthier,
2012. Isolation, identification, and technological
characterization of wild leuconostocs and
lactococci for traditional Raib type milk
fermentation. Dairy Sci. Technol., 92(3): 249264.
Benmechernene, Z., I. Fernandez, M. Kihal, K.
Böhme, P. Calo-Mata and J.B. Velazquez, 2013.
Recent Patents on Bacteriocins: Food and
Biomedical Applications. Recent Patents on
DNA & Gene Sequences, 7: 66-73
Bissonnette, F., S. Labrie, H. Deveau, M.
Lamoureux
and
S.
Moineau,
2000.
Characterization of mesophilic mixed starter
12.
13.
14.
15.
16.
17.
18.
19.
cultures used for the manufacture of aged
Cheddar cheese. J. Dairy Sci., 83: 620-627.
Boumehira, A.Z., A. Mami, J.E. Henni and M.
Kihal, 2011. Identification and Characterization
of Functional and Technological Lactobacillus
plantarum Strains Isolated from Raw Goat and
Camel Milk Collected in Algeria. J. Pure Appl.
Microbiol., 5(2): 553-566.
Cardamone, L., A. Quiberoni, D.J. Mercanti,
M.E. Fornasari, J.A. Reinheimer and D.M.
Guglielmotti,
2011.
Adventitious
dairy
Leuconostoc
strains
with
interesting
technological and biological properties useful
for adjunct starters. Dairy Sci. & Technol., 91:
457-470.
Carr, F.J. D. Chill and N.R. Maida, 2002. The
lactic acid bacteria: With literature Survey. Curr.
Rev. Microbiol., 28(4): 281-370.
Djadouni, F. and M. Kihal, 2012. Antimicrobial
Activity of Lactic Acid Bacteria and the
Spectrum of their Biopeptides Against Spoiling
Germs in Foods. Braz. Arch. Biol. Technol.,
55(3): 435-443.
Djadouni,
F.
and
M.
Kihal,
2013.
Characterization and determination of the factors
affecting
anti-listerial bacteriocins from
Lactobacillus plantarum and Pediococcus
pentosaceus isolated from dairy milk products .
Afr. J. Food Sci., 7(2): 35-44.
Drici, H., C. Gilbert, M. Kihal and D. Atlan,
2009. Atypical citrate-fermenting Lactococcus
lactis strains isolated from dromedary’s milk. J.
Appl. Microbiol., 108(2): 647-657.
Dworkin, M., S. Falkow, E. Rosenberg, K.H.
Schleifer and E. Stackebrandt, 2006. The
prokaryotes, A Handbook on the Biology of
Bacteria Volume 4: Bacteria: Firmicutes,
Cyanobacteria 6.Edition, Spinger. Singapore.,
pp: 205-311.
Facklam, R., D. Hollis and M.D. Collins, 1989.
Identification of Gram-Positive Coccal and
Cocobacillary Vancomicin-Resistant Bacteria. J.
Clin. Microbiol., 27(4): 724-730.
Gonzalez, E.T., H. Sandoval, N. Sacristan, J.M.
Castron, J.M. Fresno and M.E. Tornadja, 2007.
Identification of lactic acid bactéreria isolated
from genestoso cheese throughout riphing and
study of their antimicrobial activity. Food
Control, pp: 716-722.
Guessas, B., F. Adjoudj, M. Hadadji et M. Kihal,
2012. Isolation and Identification of Lactic Acid
Bacteria from Dhan, a Traditional Butter and
Their Major Technological Traits. World Appl.
Sci. J. 17(4): 480-488.
Guiraud, J.P., 2003. Bactérie lactique in :
microbiologie alimentaire, ed dunod, paris, pp:
89-297.
Joffin, J.N. and G. Leyral, 1996. Microbiologie
techniques, doc, bordeaux, pp: 75-239.
1006
Adv. Environ. Biol., 7(6): 999-1007, 2013
20. Kempler, G.M. and L.L. Mac Kay, 1980.
improved medium for detection of citrate
fermenting
Streptococcus
lactis
subsp.
diacetylactis .J. Appl. Environ. Microbiol., (39):
927-956.
21. Kihal, M., H. Prevost, D. E. Henni, Z.
Benmechernene and C. Diviès, 2009. Carbon
dioxide
production
by
Leuconostoc
mesenteroîdes grown in single and mixed culture
with Lactococcus lactis in skim milk. Scientific
Research and Essay, 4(11): 1348-1353.
22. Kihal, M., H. Prevost, M.E. Lhotte, D.Q. Huang
and C. Divies, 1996. Instability of plasmidencoded citrate permease in Leuconostoc. Lett.
Appl. Microbiol., 22 : 219-223.
23. Kostinek, M., I. Specht, V.A. Edward, C. Pinto,
M. Engonlety, C. Sossa, C. Dortu and P.
Thonart, 2007. Caracterisation and biochemical
properties of predominant lactic acid bacteria
from fermenting cassava for selection as starter
cultures. International Journal of Food
Microbiology, 114: 342-351.
24. Laouabdia, N.S., A. Badis, J. Guetarni, R.
Ouzroute and M. Kihal, 2007. Caractéruisation
phenotypique of lactic acid bacteria isolées from
believed milk of goat of two caprine populations
Local Arabia and Kabile.. Journal of Animal and
Veterinary Advance., 6(12): 1474-1481.
25. Leroy, F. and L. Devuyst, 2004. Lactic acid
bacteria as functional starter cultures for the
food fermentation industry. Trends Food Sci
Technol., 15: 67-78.
26. Liu, M., J.R. Bayjanov, B. Renckens, A. Nauta,
R.J Siezen, 2010. The proteolytic system of
lactic acid bacteria revisited: a genomic
comparison. BMC Genomics, 11(36): 2-15.
27. Mami, A., J.E. Henni and M. Kihal, 2012.
Screening of autochthonous Lactobacillus
species from Algerian raw gaot's milk for the
production of bacteriocin-like compounds
against Staphylococcus aureus. Afr. J.
Microbiol. Res., 6(12): 2888-2898.
28. Malek, F., B. Moussa Boudjemaa, A. AouarMétri and M. Kihal, 2013. Identification and
genetic diversity of Bacillus cereus strains
isolated from a pasteurized milk processing line
in Algeria. Dairy Sci. Technol., 93(1): 73-82.
29. Masle, I. and F. Morgan, 2001. Aptitude du lait
de chevre à l'acidification par les ferments
lactiques, facteurs de variation liés à la
composition du lait, Ed INRA, EDP Science
Lait., 81: 561-569.
30. Mathot, A.G., M. Kihal, H. Prevost and C. Divies,
1994. Selective enumeration of Leuconostoc on
vancomycine agar media. Inter. Dairy. J. 4(5):
459-469.
31. Matos, J., M. Nardi, H. Kumura and V. Monnet,
1998. Genetic characterization of pepP, which
encodes an aminopeptidase P whose deficiency
does not affect Lactococcus lactis growth in
32.
33.
34.
35.
36.
37.
38.
39.
40.
41.
42.
milk, unlike deficiency of the X-prolyl
dipeptidyl aminopeptidase. Appl. Environ.
Microbiol., 64: 4591-4595.
Mayeux, J.V., W.W.E. Sandine and P.R. Elliker,
1962. A selective medium for detecting
Leuconostoc organisms in mixed strain starter,
cultures. J. Dairy. Sci., (45): 655-656.
Merzouk, Y., W. Chahrour, K. Zarour, A.
Zergui, N. Saidi, J.E. Henni and M. Kihal, 2013.
Physico-chemical and Microbiological Analysis
of Algerian Raw Camel’s Milk and
Identification of Predominating Thermophilic
Lactic Acid Bacteria. Journal of Food Science
and Engineering., 3: 55-63.
Moulay, M., H. Aggad, Z. Benmechernene, B.
Guessas, D.E. Henni and M. Kihal, 2006.
Cultivable Lactic Acid Bacteria Isolated from
Algerian Raw Goat’s Milk and Their prteolytic
Activity. World J. Dairy Food Sci., 1(1): 12-18.
Nokuthula, F.K., I. Geornaras, A. Honly and
W.J. Hastings, 2004. Characterization and
determination of origin of lactic acid bacteria
from a soghum based fermented wearing food
by analysis of soluble proteins and amplified
fragment leught polymorphism finger printing.
Appl. Environ. Microbiol., 66(3): 1084-1092.
Sajur, S.A., F.M. Saguir and M.C.M. Nadra,
2007. Effect of dominant specie of lactic acid
bacteria from tomato on natural microflora
development in tomato purée. Food Control.,
18: 549- 600.
Siezen, R.J., B. Renckens, I. van Swam, S.
Peters, R. van Kranenburg, M., Kleerebezem
and W.M. de Vos, 2005. Complete sequences of
four plasmids of Lactococcus lactis subsp.
cremoris SK11 reveal extensive adaptation to
the dairy environment. Appl. Environ.
Microbiol., 71: 8371-8382.
Thomas, T.D., 1973. Agar medium for
differentiation of Streptococcus cremoris from
the other bacteria. N. Z. J. Dairy. Sci. Technol,
(8): 70-71.
Tserovsk, L., S.Stanka and Y. Tanye, 2004.
Identification of lactic acid bacteria isolated
form katyk, goat’s milk and cheese. J. Cult.
Collect. Nation. Bank Indust. Micro. Cell Cult.
l3: 48-52.
Varmanen, P., T. Rantanen and A. Palva, 1996.
An operon from Lactobacillus helveticus
composed of a proline iminopeptidase gene
(pepI) and two genes coding for putative
members of the ABC transporter family of
proteins. Microbiology., 142(12): 3459-3468.
Varmanen, P., T. Rantanen, A. Palva and S.
Tynkkynen, 1998. Cloning and characterization
of a prolinase gene (pepR) from Lactobacillus
rhamnosus. Appl. Environ. Microbiol., 64: 18311836.
Vasala, A., J. Panula and P. Neubauer, 2005.
Efficient lactic acid production from high salt
1007
Adv. Environ. Biol., 7(6): 999-1007, 2013
containing dairy by-products by Lactobacillus
salivarius ssp. salicinius with pre-treatment by
proteolytic microorganisms. J. Biotechnol., 117:
421-431.
43. Zarour, K., Z. Benmechernene, M. Hadadji, B.
Moussa-Boudjemaa, D. J. Henni and M. Kihal,
2012.
Bioprospecting
of
Leuconostoc
mesenteroides strains isolated from Algerian raw
camel and goat milk for technological properties
useful as adjunct starters. Afr. J. Microbiol. Res.,
6(13): 3192-3201.
44. Zarour, K.., Z. Benmechernene, M. Hadadji, B.
Moussa-Boudjemaa, J.E. Henni and M. Kihal,
2013. Caractérisation microbiologique et
technologique des espèces de Leuconostoc
mesenteroïdes isolées du lait cru de chèvre et de
chamelle d’Algérie. Revue « Nature &
Technologie ». B- Sciences Agronomiques et
Biologiques, 08: 39-47.