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Fifteen strains of extremely halophilic bacteria were isolated from fish sauce (nam-pla) collected in Thailand at various stages of the fish-fermentation process. The isolates were strictly aerobic, spore-forming, Gram-positive rods. They grew optimally in the presence of 20-26 % NaCl. The cell-wall peptidoglycan contained meso-diaminopimelic acid. The predominant menaquinone was MK-7. The major cellular fatty acids were anteiso-C(15 : 0) and anteiso-C(17 : 0). Polar lipid analysis revealed the presence of phosphatidylglycerol, diphosphatidylglycerol and two unidentified glycolipids. The DNA G+C content was 42.1-43.1 mol%. On the basis of the 16S rRNA gene sequence, a representative strain, PS11-2(T), was found to be closely related to Lentibacillus juripiscarius JCM 12147(T) (97.3 % similarity). The 15 strains were included in the same species on the basis that the levels of DNA-DNA relatedness with strain PS11-2(T) were greater than 70 %. They could be distinguished from L. juripiscarius and other Lentibacillus species on the basis of several phenotypic characteristics and low levels of DNA-DNA relatedness (</=19.4 %). Therefore, the strains represent a novel species of the genus Lentibacillus, for which the name Lentibacillus halophilus sp. nov. is proposed. The type strain is PS11-2(T) (=JCM 12149(T)=TISTR 1549(T)=PCU 240(T)).
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Lentibacillus halophilus sp. nov., from fish sauce in
Thailand
Somboon Tanasupawat,
1
Amnat Pakdeeto,
1
Sirilak Namwong,
1
Chitti Thawai,
1
Takuji Kudo
2
and Takashi Itoh
2
Correspondence
Somboon Tanasupawat
Somboon.T@chula.ac.th
1
Department of Microbiology, Faculty of Pharmaceutical Sciences, Chulalongkorn University,
254 Phayathai Road, Wangmai, Pathumwan, Bangkok 10330, Thailand
2
Japan Collection of Microorganisms, RIKEN BioResource Center, 2-1 Hirosawa, Wako-shi,
Saitama 351-0198, Japan
Fifteen strains of extremely halophilic bacteria were isolated from fish sauce (nam-pla) collected
in Thailand at various stages of the fish-fermentation process. The isolates were strictly aerobic,
spore-forming, Gram-positive rods. They grew optimally in the presence of 20–26 % NaCl. The
cell-wall peptidoglycan contained meso-diaminopimelic acid. The predominant menaquinone was
MK-7. The major cellular fatty acids were anteiso-C
15 : 0
and anteiso-C
17 : 0
. Polar lipid analysis
revealed the presence of phosphatidylglycerol, diphosphatidylglycerol and two unidentified
glycolipids. The DNA G+C content was 42?1–43?1 mol%. On the basis of the 16S rRNA gene
sequence, a representative strain, PS11-2
T
, was found to be closely related to Lentibacillus
juripiscarius JCM 12147
T
(97?3 % similarity). The 15 strains were included in the same species
on the basis that the levels of DNA–DNA relatedness with strain PS11-2
T
were greater than
70 %. They could be distinguished from L. juripiscarius and other Lentibacillus species on the basis
of several phenotypic characteristics and low levels of DNA–DNA relatedness (¡19?4 %).
Therefore, the strains represent a novel species of the genus Lentibacillus, for which the name
Lentibacillus halophilus sp. nov. is proposed. The type strain is PS11-2
T
(=JCM 12149
T
=TISTR
1549
T
=PCU 240
T
).
Moderately halophilic, endospore-forming, rod-shaped
bacteria are widely distributed in environments containing
high NaCl concentrations, suc h as saline lakes and fish
sauce. They are a diverse group of bacteria belonging to the
genera Bacillus, Halobacillus, Virgibacillus, Filobacillus,
Oceanobacillus, Lentibacillus and Pontibacillus (Ventosa
et al., 1989; Spring et al., 1996; Heydrickx et al., 1998;
Arahal et al., 2000; Schlesner et al., 2001; Lu et al., 2001;
Heyrman et al., 2003; Yoon et al., 2002, 2004; Lim et al.,
2005a). The genus Lentibacillus, which forms a phylogene-
tically coherent group , currently comprises four species:
Lentibacillus salicampi, L. juripiscarius, L. salarius and L.
lacisalsi (Yoon et al., 2002; Namwong et al., 2005; Jeon et al.,
2005; Lim et al., 2005b). In this paper, we report the isolation
and identification of some novel extremely halophilic strains
representing a novel Lentibacillus species.
Fish-sauce samples were collected from fish-sauce factories
in Thailand during the early, middle and late stages of the
fermentation process. Halophilic bacteria were isolated
from the samples by using the spread plate technique on
agar plates of JCM medium no. 168 (containing, l
21
, 200 g
NaCl, 5 g Casamino acids, 5 g yeast extract, 1 g glutamic
acid, 2 g KCl, 3 g trisodium citrate, 20 g MgSO
4
.7H
2
O,
36 mg FeCl
2
.4H
2
O, 0?36 mg MnCl
2
.4H
2
O and 20 g agar;
pH 7?2] with incubation at 37
u
C for 7 days. Liquid cultures
were cultivated in Erlenmeyer flasks containing the same
medium without agar and were incubated on a rotary
shaker. All media contained 20 % (w/v) NaCl, except those
used to investigate NaCl tolerance. Cell shape, cell size and
cell arrangement were examined on JCM medium no. 168
agar at 37
u
C for 5 days. The Hucker–Conn modification
was used for Gram staining (Hucker & Conn, 1923). Spore
formation was examined on Gram-stained specimens.
Critical-point-dri ed cells were observed under a scanning
electron microscope. Flagella were examined as described by
Forbes (1981) and observed by transmission electron
microscopy. Catalase activity, oxidase activity and the
hydrolysis of aesculin were investigated as described by
Barrow & Feltham (1993), while urease activity and the
hydrolysis of gelatin, casein, starch, Tween 80, tyrosine,
The GenBank/EMBL/DDBJ accession number for the 16S rRNA gene
sequence of strain PS11-2
T
is AB191345.
A scanning electron micrograph of sporulating cells of strain PS11-2
T
,
a thin-layer chromatogram of polar lipids and details of the cellular fatty
acids and levels of DNA–DNA relatedness of the novel strains and
related taxa are available as supplementary material in IJSEM Online.
63997
G
2006 IUMS Printed in Great Britain 1859
International Journal of Systematic and Evolutionary Microbiology (2006), 56, 1859–1863 DOI 10.1099/ijs.0.63997-0
phenylalanine, xanthine and hypoxanthine were tested as
described by Namwo ng et al. (2005). Arginine hydrolysis
was investigated by using the mediu m reported by Thornley
(1960). Acid production from carbohydrate was determined
in the medium described by Leifson (1963), supplemented
with 20 % NaCl. Growth under anaerobic conditions on
agar plates with or without KNO
3
(1 %, w/v) was performed
in a Gaspak (BBL) anaerobic jar. Growth at various tem-
peratures (10–50
u
C), pH values (5, 6, 7, 7 ?5, 8 and 9) and
NaCl concentrations (0–30 %, w/v) was tested by using JCM
medium no. 168 as a basal medium (Namwong et al., 2005).
The diaminopimelic acid in the peptidoglycan and the
menaquinone composition were determined as described
previously (Komagata & Suzuki, 1987). Polar lipids were
investigated according to the methods of Minnikin et al.
(1984) and Albert et al. (2005). A loop of cell mass was used
for the extraction and quantitative analysis of the cellular
fatty acids by means of the Microbial Identification System
(MIDI) (Sasser, 1990; Ka
¨
mpfer & Kro ppenstedt, 1996).
DNA was isolated from cells grown in JCM medium no. 168
broth and purified according to the method of Saito &
Miura (1963). The DNA G+C content was determined
by the method of Tamaoka & Komagata (1984), using
reversed-phase HPLC. DNA–DNA hybridization was con-
ducted in microdilution-well plates, as reported by Ezaki
et al. (1989), and was detected by using the colorimetric
method described by Tanasupawat et al. (2000). The 16S
rRNA gene of the isolate was amplified, purified and
sequenced as described previously (Seearunruangchai et al.,
2004). The sequence determined (1410 bases) was aligned
with selected sequences (obtained from the GenBank/
EMBL/DDBJ database) by using
CLUSTAL W, version 1.81
(Thompson et al., 1994). The alignment was manually
edited to remove gaps and ambiguous nucleotides prior to
the construction of the phylogenetic tree. The phylogenetic
tree was constructed by using the neigh bour-joining method
(Saitou & Nei, 1987) in
MEGA, version 2.1 (Kumar et al.,
2001). The confidence values of branches of the phylo-
genetic tree were determined using bootstrap analyses
(Felsenstein, 1985) based on 1000 resamplings.
Fifteen aerobic, extremely halophilic bacteria were isolated
from the fish-sauce samples, and were characterized accord-
ing to their morphological, cultural, physiological and
Table 1. Differential characteristics of Lentibacillus species
Strains: 1, PS11-2
T
;2,L. juripiscarius JCM 12147
T
(data from Namwong et al., 2005); 3, L. salarius KCTC 3911
T
(Jeon et al., 2005); 4, L.
salicampi JCM 11462
T
(Yoon et al., 2002; Namwong et al., 2005); 5, L. lacisalsi KCTC 3915
T
(Lim et al., 2005b). +, Positive; 2 , negative;
W, weak; NA, no data available.
Characteristic 1 2 3 4 5
Spore shape Spherical Oval Spherical/oval Spherical/oval Spherical
Motility + 2 +++
Colony diameter (mm) 0?2–0?61?3–3?2
NA 0?2–1?3 NA
Maximum temperature for growth (uC) 42 45 50 40 40
NaCl range for growth (%) 12–30 3–30 1–20 3–25 5–25
Oxidase ++ 2 ++
Nitrate reduction 2 ++ + +
Hydrolysis of:
Aesculin 22 + 22
Casein 2 + 2 + 2
Tween 80 2 + 2 + 2
Acid production from:
Cellobiose 22
NA W NA
D
-Glucose 2 ++ + 2
D-Galactose 22 NA W NA
D
-Fructose 2 ++ 2 +
Maltose 22 +
W 2
D-Mannitol 22 W 22
D-Mannose 22 + W 2
D-Ribose 2 ++ 2 +
Salicin 22 2
W 2
Sucrose 2
WNA 2 NA
D
-Trehalose 22 W 22
D-Xylose 2 ++ 2 W
DNA G+C content (mol%) 42 43 43 42 44
1860 International Journal of Systematic and Evolutionary Microbiology 56
S. Tanasupawat and others
biochemical properties. The results are listed in the species
description and Table 1. All 15 strains contained meso-
diaminopimelic acid as the diagnostic diamino acid in the
cell-wall peptidoglycan. The menaquinones, cellular fatty
acids and polar lipids of four of these strains, namely
PS11-2
T
, CB0-1, DS26-3 and DB9-1, were analysed. The
four strains contained the following: MK-7 as a major
menaquinone; anteiso-C
15 : 0
(58?0–62?7 %), anteiso-C
17 : 0
(24?6–33?8 %), C
16 : 0
(1?7–3?0 %), iso-C
15 : 0
(2?7–7?8%)
and iso-C
16 : 0
(1?8–2?3 %) as the cellular fatty acids (see
Supplementary Table S1 available in IJSEM Online); and
phosphatidylglycerol, diphosphatidylglycerol and two un-
identified glycolipids as major polar lipids (see Supple-
mentary Fig. S1 available in IJSEM Online).
The DNA G+ C content of the isolates (13 strains) ranged
from 42 ?1to43?1 mol%. In the neighbour-joining phylo-
genetic tree based on 16S rRNA gene sequences, strain
PS11-2
T
was positioned in a monophyletic cluster consisting
of the members of the genus Lentibacillus (Fig. 1). The
same top ology was obtained by applying the maximum-
parsimony method (result not shown). The 16S rRNA
gene sequence similarity values between PS11-2
T
and L.
juripiscarius JC M 12147
T
, L. salarius KCTC 3911
T
, L.
lacisalsi KCTC 3915
T
and L. salicampi JCM 11462
T
were
97?3, 95?5, 95?4 and 95?3 %, respectively. Furthermore,
strain PS11-2
T
showed 93?3–94?2 % 16S rRNA gene
sequence similarity to members of the genus Virgibacillus.
Hybridization studies revealed high levels of DNA–DNA
relatedness between PS11-2
T
and PB7-3 and to the other
isolates (>70 %), but only low levels with respect to L.
salicampi JCM 11462
T
(4?9–19?4%) and L. juripiscarius
JCM 12147
T
(17?0–17?1 %), as shown in Supplementary
Table S2 (available in IJSEM Online).
The 16S rRNA gene sequence-based phylogenetic analysis
clearly indicated that representative strain PS11-2
T
belongs
to the genus Lentibacillus. The chemotaxonomic properties
(i.e. diamino acid content in the peptidoglycan, menaqui-
none content, the polar lipids and the cellular fatty acid
profiles) of the isolates were in accordance with those of the
genus Lentibacillus (Yoon et al., 2002; Namwong et al., 2005;
Jeon et al., 2005; Lim et al., 2005b). The fatty acid profiles of
the four strains examined were qualitatively similar to those
of other Lentibacillus species, although the levels of iso-C
14 : 0
and iso-C
16 : 0
were significantly lower than those of the
other Lentibacillus species reported (¡0?7 % iso-C
14 : 0
and
1?8–2?3 % iso-C
16 : 0
for the isolates; 5?7–13?9 % iso-C
14 : 0
and 16?3–26?5 % iso-C
16 : 0
for Lentibacillus species). When
the cellular fatty acid profile of L. juripiscarius JCM 12147
T
grown on JCM medium no. 168 (containing 20 % NaCl) was
compared with the profile from cells grown on the Lenti-
bacillus medium (JCM medium no. 377, containing 10 %
NaCl), the levels of iso-C
14 : 0
and iso-C
16 : 0
were signifi-
cantly lower (2?1 and 6?3 %, respectively: see Supplementary
Table S1 in IJSEM Online). Thus, the levels of iso-C
14 : 0
and
iso-C
16 : 0
were affected by NaCl concentration during growth.
The morphological, cultural, physiological and biochemical
characteristics that differentiate the isolates from Lentibacillus
species are shown in Table 1. It is noteworthy that all of the
isolates are extreme halophiles requiring at least 12 % NaCl
for growth. In addition, the isolates do not reduce nitrate and
do not produce acid from sugars, unlike Lentibacillus species.
The 16S rRNA gene sequence of PS11-2
T
shows a relatively
Fig. 1. Phylogenetic tree, based on 16S
rRNA gene sequences, showing the relation-
ships between strain PS11-2
T
and related
bacterial species. The branching pattern was
generated by the neighbour-joining method.
Bootstrap percentages above 75 %, based
on 1000 replications, are shown at the
nodes. Bar, 1 substitution per 100 nucleo-
tide positions.
http://ijs.sgmjournals.org 1861
Lentibacillus halophilus sp. nov.
high level of similarity to that of the type strain of L.
juripiscarius (97?3 %). However, the low levels of DNA–DNA
relatedness (¡19?4 %) demonstrate that the novel strains do
not belong to the species L. juripiscarius. On the other hand,
inclusion of the 15 strains in the same species is supported by
the fact that they share the same phenotypic properties and
demonstrate high levels of DNA–DNA relatedness to each
other (>70 %) (Wayne et al., 1987). Therefore, these 15
isolates represent a novel species in the genus Lentibacillus,
for which we propose the name Lentibacillus halophilus sp.
nov. The type strain is PS11-2
T
(=JCM 12149
T
=TISTR
1549
T
=PCU 240
T
).
Description of Lentibacillus halophilus sp. nov.
Lentibacillus halophilus (ha.lo9phi.lus. Gr. n. hals, halos salt;
Gr. adj. philos loving; N.L. masc. adj. halophilus salt-loving).
Cells are Gram-positive, aerobic, motile rods and are mostly
0?4–0?6
mm wide and 1?0–3? 0 mm long. Longer cells (up to
6 mm) or short filaments are observed. Spherical endospores
are formed terminally in swollen sporangia (see Supple-
mentary Fig. S2 available in IJSEM Online). Colonies are
white to cream, low-convex or raised, smooth and circular
(0?1–0?8 mm in diameter). Catalase- and oxidase-positive.
Urease-negative. Growth occurs between 15
u
C (weakly) and
42
u
C (optimum, 30–37
u
C) but not at 10, 45 or 50
u
C.
Growth is observed between pH 6 and 8 (optimum,
pH 7?0–7?5) but not at pH 5 or 9. Extremely halophilic,
growing in the presence of 12–30 % (w/v) NaCl but not at
or below 10 % (w/v) NaCl (optimum, 20–26 % NaCl, w/v).
Anaerobic growth is not observed in the presence of 1 %
KNO
3
(w/v) or in other media. Aesculin, arginine, casein,
gelatin, Tween 80, tyrosine, starch, phenylalanine, xanthine
and hypoxanthine are not hydrolysed. Acid is not produced
from
D-glucose, glycerol, D-ribose, D-xylose, L-arabinose,
cellobiose, D-fructose, D-galactose, lactose, maltose, D-
mannitol,
D-mannose, D-melibiose, D-melezitose, myo-
inositol, raffinose, L-rhamnose, salicin, sorbitol, sucrose or
D-trehalose. Contains meso-diaminopimelic acid as the
diagnostic diamino acid in the cell-wall peptidoglycan. MK-
7 is the major menaquinone. The fatty acid profile consists
of anteiso-C
15 : 0
(58?0–62?7 %), anteiso-C
17 : 0
(24?6–33?8%),
iso-C
15 : 0
(2?4–7?8 %), C
16 : 0
(1?7–3?0 %), iso-C
16 : 0
(1?8–2?3 %), iso-C
17 : 0
(0?5–1?4 %), C
14 : 0
(0–0?9 %), iso-
C
14 : 0
(0–0?7 %), C
15 : 0
(0–0?6 %) and C
12 : 0
(0–0?5 %).
Phosphatidylglycerol, diphosphatidylglycerol and two uni-
dentified glycolipids are predominant in the polar lipid
profile. The DNA G+C content is 42?1–43?1 mol% (type
strain, 42?4 mol%).
The type strain, PS11-2
T
(=JCM 12149
T
=TISTR 1549
T
=
PCU 240
T
), was isolated from a fish-sauce fermentation in
Thailand.
Acknowledgements
We are very grateful to Sindhu Samuth Fish Sauce Factory (Squid
Brand) Ltd (Samutprakarn), Thai Fish Sauce Factory (Squid Brand)
Co., Ltd (Samutsongkram) and Pichai Fish Sauce Co., Ltd (Chonburi)
for providing the samples. This study was supported, in part, by
Ratchadapiseksomphot Research Grant, Chulalongkorn University
(2002).
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http://ijs.sgmjournals.org 1863
Lentibacillus halophilus sp. nov.

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... At the time of writing, the genus contained 16 validly named species, as reported on the LSPN website (www.bacterio.net/lentibacillus.html), including the recently described Lentibacillus sediminis 0W14 T from a marine saltern [2], Lentibacillus amyloliquefaciens LAM0015 T from a saline sediment [3], Lentibacillus kimchii K9 T from kimchi [4], Lentibacillus populi WD4L-1 T from a poplar tree [5], Lentibacillus garicola MJ1 T from a Korean fermented anchovy sauce [6], Lentibacillus jeotgali Grbi T from traditional Korean fermented seafood [7], Lentibacillus persicus Amb31 T from a saline lake [8], Lentibacillus salinarum AHS-1 T from a marine solar saltern in the Republic of Korea [9], Lentibacillus salis BH113 T [10] and Lentibacillus halodurans 8-1 T from a salt lake in PR China [11], Lentibacillus kapialis PN7-6 T from fermented shrimp paste [12], Lentibacillus halophilus PS11-2 T [13] and Lentibacillus juripiscarius IS40-3 T [14] from fish sauce, Lentibacillus lacisalsi BH260 T from a saline lake [15], and Lentibacillus salarius BH139 T from saline sediment in PR China [16]. In the course of screening for the diversity of moderately halophilic bacteria from Thai fermented food, strain SSKP1-9 T was isolated from Ka-pi collected from Samut Sakhon Province, Thailand. ...
... Major polar lipids were phosphatidylglycerol and diphosphatidylglycerol and there were minor amounts of unidentified lipids (L1-L4), an unidentified phospholipid (PL1) and glycolipid (Fig. S7). The predominant isoprenoid quinone was MK-7, the same as in L. juripiscarius TISTR 1535 T and L. halophilus TISTR 1549 T [13,14]. ...
Article
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A Gram-stain-positive, aerobic, spore-forming, moderately halophilic bacterium, SSKP1-9T, was isolated from traditional salted shrimp paste (Ka-pi) produced in Samut Sakhon Province, Thailand. This strain grew optimally at 37-40 °C, pH 7.0 and in the presence of 8-16 % (w/v) NaCl. The 16S rRNA gene sequence similarity values between strain SSKP1-9T and Lentibacillus juripiscarius TISTR 1535T and Lentibacillus halophilus TISTR 1549T were 98.7 and 97.2 %, respectively. Based on 16S rRNA gene sequence similarity, strain SSKP1-9T represents a distinct novel species, as shown by phenotypic traits, DNA-DNA hybridization and average nucleotide identity values. In addition, the whole-cell protein profile confirmed the novelty of the taxon. The genomic DNA G+C content was 44.6 mol%. The major isoprenoid quinone was MK-7. The cell-wall peptidoglycan contained meso-diaminopimelic acid. Polar lipid analysis revealed the presence of phosphatidylglycerol, diphosphatidylglycerol, four unidentified lipids, an unidentified phospholipid and an unidentified glycolipid. The major cellular fatty acids were anteiso-C15 : 0, anteiso-C17 : 0 and iso-C16 : 0. The results of phenotypic and chemotaxonomic characteristics and whole-genome analysis support that strain SSKP1-9T represents a novel species of Lentibacillus, for which the name Lentibacilluslipolyticus sp. nov. is proposed. The type strain is SSKP1-9T (=JCM 32625T=TISTR 2597T).
... Lentibacillus members are mainly characterized as moderate or extreme halophiles with menaquinone-7 (MK-7) as the predominant menaquinone. Thus far, 19 species belonging to the genus Lentibacillus have been identified in a number of salty enviroments such as salt lakes Lim et al., 2005;Yuan et al., 2007;Lee et al., 2008b;Sanchez-Porro et al., 2010), salt fileds (Yoon et al., 2002;Lee et al., 2008a;Wang et al., 2016;Guo et al., 2017), fermented food (Namwong et al., 2005;Tanasupawat et al., 2006;Pakdeeto et al., 2007;Jung et al., 2010Jung et al., , 2015Oh et al., 2016a;Sundararaman et al., 2018;Booncharoen et al., 2019), poplar trees (Sun et al., 2016), and human stools (Senghor et al., 2017). At the time of writing, a total of 9 genomes related to Lentibacillus species are published in NCBI, genome properties are indicated as following genome size (3,146,926,733 bp), CDSs (3,045-5,053), rRNA genes (10-30), and tRNA genes (50-86). ...
... from Namwong et al. (2005) b Data fromTanasupawat et al. (2006) c Data fromBooncharoen et al. (2019) d PG, phosphatidylglycerol; DPG, diphosphatidylglycerol; APL, unidentified aminophospholipid; GL, unidentified glycolipid; PL, unidentified phospholipid; L, unidentified lipid. e DNA G + C content of strains was calculated from the respective genome. ...
Article
Two bacterial strains designated NKC220-2T and NKC851-2 were isolated from commercial kimchi from different areas in Korea. The strains were Gram-positive, aerobic, oxidase-and catalase-positive, rod-shaped, spore-forming, non-motile, and halophilic bacteria. Both strains grew without NaCl, unlike type species in the genus Lentibacillus. The optimal pH for growth was 8.0, higher than that of the type species in the genus Lentibacillus, although growth was observed at pH 5.5-9.0. 16S rRNA gene sequence-based phylogenetic analysis indicated that the two strains (99.3-99.9% similarity) are grouped within the genus Lentibacillus and most closely related to Lentibacillus juripiscarius IS40-3T (97.4-97.6% similarity) isolated from fish sauce in Thailand. OrthoANI value between two novel strains and Lentibacillus lipolyticus SSKP1-9T (79.5-79.6% similarity) was far lower than the species demarcation threshold. Comparative genomic analysis displayed differences between the two strains as well as among other strains belonging to Lentibacillus. Furthermore, each isolate had strain-specific groups of orthologous genes based on pangenome analysis. Genomic G + C contents of strains NKC-220-2T and NKC851-2 were 41.9 and 42.2 mol%, respectively. The strains contained meso-diaminopimelic acid in their cell walls, and the major menaquinone was menaquinone-7. Phosphatidylglycerol, diphosphatidylglycerol, and an unidentified glycolipid, aminophospholipid, and phospholipid were the major polar lipid components of both strains. The major cellular fatty acids of the strains were anteiso-C15:0 and an-teiso-C17:0. Based on phenotypic, genomic, phylogenetic, and chemotaxonomic features, strains NKC220-2T and NKC851-2 represent novel species of the genus Lentibacillus, for which the name Lentibacillus cibarius sp. nov. is proposed. The type strain is NKC220-2T (= KACC 21232T = JCM 33390T).
... One example is the Thai anchovy fish sauce nam-pla, which is made by adding two parts of fish to one part of marine salts, adding concentrated brine, and fermenting for a year. Various halophiles have been isolated from this mixture, including Lentibacillus halophilus [177], ...
... G56 T ; 2, L. juripiscarius JCM 12147 T[25]; 3, L. halophilus JCM 12149 T[26]; 4, L. salis JCM 32144 T[27]; 5, L. lacisalsi JCM 19838 T[28] All tests were performed under the same conditions. Symbol: + , positive reaction; − , negative reaction; w weakly positive, NA no data available. ...
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A Gram-positive, aerobic, motile and short rod-shaped bacterium, designated as strain G56T, was isolated from shrimp paste produced in Panjin, China. Grows in the presence of 1.0–25.0% (w/v) NaCl (optimum at 10%), pH 5.0–9.5 (optimally at 7.0) and 10–50 °C (optimally at 37 °C). Positive for catalase and oxidase activities, but lack the ability to reduce nitrate. Acids produce from d-ribose, d-xylose, d-galactose, glycerol and d-trehalose, but no acid is produced when salicin, d-mannose, d-cellobiose and l-sorbose are provided as substrates. The polar lipid extract is found to contain diphosphatidylglycerol, phosphatidylglycerol, an unknown glycolipid, and unidentified phospholipids. Fatty acids are mainly defined as anteiso-C15:0 (69.7%) and anteiso-C17:0 (23.3%). The G+C content of its DNA is 44.7 mol%. The draft genome of strain G56T is 3,209,087 bp in length and the average nucleotide identity value (ANI) and the digital DNA-DNA hybridization (DDH) values between strain G56T and L. juripiscarius JCM 12147T is 78.41% and 22.0%, respectively. Polyphasic taxonomic analysis classified strain G56T as a novel species in the genus Lentibacillus, and therefore, we named it as Lentibacillus panjinensis sp. nov..
... One example is the Thai anchovy fish sauce nam-pla, which is made by adding two parts of fish to one part of marine salts, adding concentrated brine, and fermenting for a year. Various halophiles have been isolated from this mixture, including Lentibacillus halophilus [177], ...
Chapter
Extremophiles occupy habitats characterized by extremes of temperature, pH, salinity, hydrostatic pressure, ionizing radiation, dessication and high concentrations of metals. Most are Bacteria or Archaea and possess numerous molecular adaptations that allow them to withstand these conditions. These include production of thermostable or cold-stable proteins, enzymes that are stable at pH extremes, altered membrane composition, production of compatible solutes, and secondary metabolites that form radioprotective screens. These molecules have been applied in various industries, including food production and as medicines or components of medicines. Some extremophiles are pathogenic, can cause food spoilage, or can be used to make fermented foods. In addition to their applications, extremophiles provide valuable information about the physiochemical boundaries of life.
... bacterio.net/lentibacillus.html). Most members of this genus are moderately halophilic, except the species Lentibacillus halophilus and Lentibacillus kimchii, which are extremely halophilic [3,4]. In this study, a novel, moderately halophilic strain, isolated from a marine saltern, was characterized using a polyphasic approach. ...
Article
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A novel, Gram-stain-positive, moderately halophilic, endospore-forming, motile, facultatively anaerobic and rod-shaped strain, designated 0W14T, was isolated from a marine saltern of Wendeng, China. Optimal growth occurred at 37 °C, pH 7.5 and with 6.0 % (w/v) NaCl. MK-7 was the sole respiratory quinone and the peptidoglycan type of 0W14T was A4βl-Orn-d-Glu. The major cellular fatty acid (>10.0 %) in strain 0W14T was anteiso-C15 : 0. The polar lipid profile of strain 0W14T consisted of diphosphatidylglycerol, phosphatidylglycerol, two unknown glycolipids and four unknown phospholipids. The genomic DNA G+C content of the strain was 44.8 mol%. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain 0W14T forms a phylogenetic lineage with members of the genus Lentibacillus within the family Bacillaceae. Based on data from the current polyphasic study, the isolate is proposed to represent a novel species of genus Lentibacillus, for which the name Lentibacillus sediminis sp. nov. is proposed. The type strain is 0W14T (=KCTC 33835T=MCCC 1H00171T).
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The extremely halophilic archaeon Halobacterium salinarum strain ETD5 was previously isolated from the solar saltern of Sfax (Tunisia) and shown to encode and express halocin S8. The Hbt. salinarum ETD5 culture supernatant was shown here to exhibit high antimicrobial activity against several halophilic archaea and bacteria of different genera, showing a cross-domain inhibition. The antimicrobial activity was destroyed by proteases, thus pointing to halocins. A bioguided purification procedure was applied using two chromatography steps and antimicrobial assays directed against Halorubrum chaoviator ETR14. In-gel screening assay showed the presence of two antimicrobial bands of approximately 8 and 14 kDa, for which characterization was investigated by N-terminal sequencing and mass spectrometry. The full-length form of halocin S8 that contains 81 amino acids and differs from the 36 amino acid short-length halocin S8 previously described from an uncharacterized haloarchaeon S8a, was identified in the 8 kDa halocin band. A novel halocin that we termed halocin S14 was found in the 14 kDa band. It exhibits amino acid sequence identities with the N-terminally truncated region of the archaeal Mn-superoxide dismutase. These results show that Hbt. salinarum ETD5 produces multiple halocins, a feature that had not been described until now in the domain Archaea.
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Bacterial communities in fish sauce, at different fermentation phases, from two large manufacturing plants (Factories MC and SQ) in Thailand, were studied. The bacterial microbiota in a fish sauce mash of >12 months were examined by 16S rRNA next-generation sequencing. We revealed the presence of 7 phyla, 14 classes, 21 orders, 31 families, and 45 genera of bacteria in the fish sauce. Bacterial communities in the sauces differed across the two factories. In Factory MC, Halanaerobium sp. was the initial dominant genus, which gradually decreased while Lentibacillus sp., Halomonas sp., and Tetragenococcus sp. began to appear in the middle phase. Diversity was the highest in the late fermentation phase. On the contrary, in Factory SQ, high diversity was observed right from the early phase till the late fermentation phase. The bacterial communities varied at different stages of fermentation as well. The most common genera were Peptostreptococcus sp., Peptoniphilus sp., Gallicola sp., Fusobacterium sp., Halanaerobium sp., and Vagococcus sp.. Alpha and beta diversity within the bacterial communities in fish sauce from the factories was also calculated. This is the first report revealing bacterial communities during industrial fermentation of Thai fish sauce. The source of raw materials and bacterial microbiota in the fermentation tank influenced the type of bacteria present throughout the process, and consequently affected the unique product characteristics.
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Len.ti.ba.cil'lus. L. adj. lentus slow; L. dim. n. bacillus small rod; N.L. masc. n. Lentibacillus slowly growing bacillus/rod. Firmicutes / “Bacilli” / Bacillales / Bacillaceae / Lentibacillus Rod-shaped cells, forming terminal endospores that swell the sporangia. Gram-variable, motile or nonmotile. Colonies are white to cream-colored, smooth and circular to slightly irregular. Catalase-positive, oxidase variable, and urease-negative. Unable to hydrolyze starch, tyrosine, or xanthine. No acid production from d-melibiose, raffinose, or l-rhamnose. Moderately to extremely halophilic. The major fatty acid is C15:0 anteiso and branched saturated fatty acids account for 95% total fatty acids. The cell-wall peptidoglycan contains meso-diaminopimelic acid at position 3 of the peptide subunit. The predominant menaquinone is MK-7. The major polar lipids are diphosphatidylglycerol and phosphatidylglycerol. DNA G + C content (mol%): 42.0–44.0. Type species: Lentibacillus salicampi Yoon, Kang and Park 2002, 2047VP.
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