International Journal of Systematic and Evolutionary Microbiology (2015), 65, 189–194
DOI 10.1099/ijs.0.064477-0
Burkholderia insulsa sp. nov., a facultatively
chemolithotrophic bacterium isolated from an
arsenic-rich shallow marine hydrothermal system
Antje Rusch,1,2 Shaer Islam,1 Pratixa Savalia3 and Jan P. Amend3,4
Correspondence
1
Antje Rusch
[email protected]
2
Department of Microbiology, Southern Illinois University Carbondale, 1125 Lincoln Drive,
Carbondale, IL 62901, USA
Center for Ecology, Southern Illinois University Carbondale, 1125 Lincoln Drive, Carbondale,
IL 62901, USA
3
Department of Earth Sciences, University of Southern California, 3616 Trousdale Parkway,
Los Angeles, CA 90089, USA
4
Department of Biological Sciences, University of Southern California, 3616 Trousdale Parkway,
Los Angeles, CA 90089, USA
Enrichment cultures inoculated with hydrothermally influenced nearshore sediment from Papua
New Guinea led to the isolation of an arsenic-tolerant, acidophilic, facultatively aerobic bacterial
strain designated PNG-AprilT. Cells of this strain were Gram-stain-negative, rod-shaped, motile
and did not form spores. Strain PNG-AprilT grew at temperatures between 4 6C and 40 6C
(optimum 30–37 6C), at pH 3.5 to 8.3 (optimum pH 5–6) and in the presence of up to 2.7 %
NaCl (optimum 0–1.0 %). Both arsenate and arsenite were tolerated up to concentrations of at
least 0.5 mM. Metabolism in strain PNG-AprilT was strictly respiratory. Heterotrophic growth
occurred with O2 or nitrate as electron acceptors, and aerobic lithoautotrophic growth was
observed with thiosulfate or nitrite as electron donors. The novel isolate was capable of N2fixation. The respiratory quinones were Q-8 and Q-7. Phylogenetically, strain PNG-AprilT belongs
to the genus Burkholderia and shares the highest 16S rRNA gene sequence similarity with
the type strains of Burkholderia fungorum (99.8 %), Burkholderia phytofirmans (98.8 %),
Burkholderia caledonica (98.4 %) and Burkholderia sediminicola (98.4 %). Differences from
these related species in several physiological characteristics (lipid composition, carbohydrate
utilization, enzyme profiles) and DNA–DNA hybridization suggested the isolate represents a novel
species of the genus Burkholderia, for which we propose the name Burkholderia insulsa sp. nov.
The type strain is PNG-AprilT (5DSM 28142T5LMG 28183T).
The betaproteobacterial genus Burkholderia comprises 85
species with validly published names at the time of writing
[LPSN (www.bacterio.net), Sept. 2014] populating two
major lineages and at least three smaller clades (SuárezMoreno et al., 2012; Estrada-de los Santos et al., 2013;
Martı́nez-Aguilar et al., 2013). Across lineages, the genus
includes plant, animal and human pathogens as well as
non-pathogenic species isolated mostly from plants and
soils, but also from freshwater sediment and an industrial
wastewater treatment system. Clinical and environmental
isolates, free-living and symbiotic lifestyles, plant-beneficial
and plant-pathogenic functions can even occur within the
same species (Vial et al., 2011; Peeters et al., 2013;
Vandamme et al., 2013; Verstraete et al., 2014). Many
members of the genus show diazotrophy and degrade a
variety of xenobiotic compounds (Coenye & Vandamme,
2003; Suárez-Moreno et al., 2012; Estrada-de los Santos
et al., 2013). Several species of the genus Burkholderia are
remarkably tolerant of metals and metalloids (Estrada-de
los Santos et al., 2011), and genes encoding arsenite oxidase
or arsenate reductase occur in 25 Burkholderia genomes
[JGI-IMG (http://img.jgi.doe.gov/), NCBI-RefSeq (http://
www.ncbi.nlm.nih.gov/refseq/); Oct. 2013]. Non-pathogenic
members of the genus are potentially relevant to agriculture
and bioremediation.
The GenBank/EMBL/DDBJ accession number for the 16S rRNA gene
sequence of strain PNG-AprilT is KF733462.
In May 2005, short (~30 cm) sediment cores were collected
by scuba divers at 30 m distance from a hydrothermal vent
in Tutum Bay (04u 059 S 153u 339 E), Ambitle Island,
Papua New Guinea. This arsenic-rich hydrothermal area at
Three supplementary tables and a supplementary figure are available
with the online Supplementary Material.
064477 G 2015 IUMS
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189
A. Rusch and others
5–10 m water depth has been described extensively as to its
geochemistry (Pichler et al., 1999; Price et al., 2007), energy
sources (Akerman et al., 2011) and microbial community
composition (Meyer-Dombard et al., 2012, 2013). The
liquid growth medium PNG-AR2 was custom-designed to
resemble resources and conditions at the sampling site,
while adding a complex carbon source and leaving out all
electron acceptors except for arsenate and small amounts
of oxygen. The composition of this medium is given in
Table S1 (available in the online Supplementary Material).
Batches of PNG-AR2 were inoculated with homogenized
sediment slurries from the top 10 cm of the core and
incubated at 30 uC. Cultures were transferred at irregular
intervals, reducing diversity by dilution and by repeated
streaking on PNG-AR2 agar, until an isolate was obtained.
The novel strain was designated PNG-AprilT and is the
only strain of its species.
DNA of strain PNG-AprilT was extracted from a pure
culture in liquid medium PNG-AR2, and three overlapping
segments of the 16S rRNA gene were amplified by PCR;
information on the PCR primers is compiled in Table S2.
Both strands were sequenced and assembled to a nearly
complete sequence of 1414 bp. Using the EzTaxon-e server
(Kim et al., 2012), strain PNG-AprilT was identified as a
member of the genus Burkholderia, sharing highest 16S
rRNA gene sequence similarity with the type strains of
Burkholderia fungorum (99.8 %), Burkholderia phytofirmans
(98.8 %), Burkholderia caledonica (98.4 %) and Burkholderia
sediminicola (98.4 %). Nearly complete 16S rRNA gene
sequences of the novel isolate and related type strains within
species of the genus Burkholderia were aligned using CLUSTAL
W 2.0 (Larkin et al., 2007), before applying the maximumlikelihood algorithm of PhyML 3.0 (Guindon et al., 2010)
with bootstrap resampling (500 replicates). The resulting
phylogenetic tree is shown in Fig. 1.
70
Growth of strain PNG-AprilT was supported by the complex media PNG-AR2 (Table S1), R-2A (Reasoner &
Geldreich, 1985) and tryptic soy (TS) broth or agar,
nutrient agar, MacConkey agar and marine broth (Difco;
diluted to 50 % or 75 %), as well as the defined media MSB
(Stanier et al., 1966) and B-litho (custom-designed, Table
S3). On R-2A agar, the isolate formed smooth, off-white to
cream colonies. Cells of strain PNG-AprilT were straight
rods of 2–3 mm length and 1 mm width, were motile by
polar flagella, stained Gram-negative and showed no spore
formation even at 40 uC.
Chemotaxonomic characterization of the novel isolate
included analyses of membrane lipids, respiratory quinones
and polar lipids. After growth on TS agar, the fatty acid
composition of cellular lipids was quantified by GC analysis
of their methyl esters, using the Sherlock Microbial ID
system (MIDI). Results obtained with strain PNG-AprilT, in
comparison with published data from closely related species
of the genus Burkholderia grown on TS medium, are shown
in Table 1. The similarity of these fatty acid profiles supports
placement of the novel isolate within the genus Burkholderia,
while specific differences in several fatty acids dissuade from
membership in any of the existing species.
From aerobic cultures in R-2A broth, cells were harvested
and freeze-dried, before quinones were extracted and
separated by TLC and subsequent HPLC (Tindall, 1990a, b).
Measurements of UV absorption detected ubiquinones Q-8
(92 %) and Q-7 (8 %). Polar lipids were extracted from
freeze-dried cells (Bligh & Dyer, 1959), separated by twodimensional TLC and stained to visualize specific functional
groups (Tindall et al., 2007). In strain PNG-AprilT, four
phospholipids, two aminophospholipids, one glycolipid,
phosphatidylglycerol and phosphatidylethanolamine were
detected (Fig. S1).
Burkholderia insulsa PNG-AprilT (KF733462)
Burkholderia fungorum LMG 16225T (AF215705)
Burkholderia phytofirmans PsJNT (AY497470)
Burkholderia bryophila LMG 23644T (AM489501)
52
Burkholderia sediminicola HU2-65WT (EU035613)
0.02
Burkholderia xenovorans LB400T (U86373)
74
95
72
Burkholderia caledonica LMG 19076T (AF215704)
72
Burkholderia dilworthii WSM3556T (HQ698908)
Burkholderia rhynchosiae WSM3937T (EU219865)
Burkholderia phenoliruptrix AC1100T (AY435213)
Burkholderia megapolitana LMG 23650T (AM489502)
Burkholderia phenazinium LMG 2247T (U96936)
Burkholderia glathei LMG 14190T (U96935)
58
82
Burkholderia rhizoxinica HKI 454T (AJ938142)
Burkholderia mallei ATCC 23344T (AF110188)
Burkholderia caryophylli ATCC 25418T (AB021423)
Pandoraea apista LMG 16407T (AF139173)
190
Fig. 1. Maximum-likelihood tree reconstructed
from nearly complete 16S rRNA gene
sequences of strain PNG-AprilT and type
strains of related species within the genus
Burkholderia, using Pandoraea apista LMG
16407T as an outgroup. Branch support (500
bootstraps) is indicated where larger than
50 %. Bar, 0.02 substitutions per site.
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Arsenic-tolerant facultatively lithotrophic Burkholderia
Table 1. Fatty acid composition of strain PNG-AprilT in
comparison to published profiles of closely related species of
the genus Burkholderia
Taxa: 1, PNG-AprilT (data from this study); 2, B. fungorum (n59,
where n is number of strains; Coenye et al., 2001); 3, B. phytofirmans
PsJNT (Sessitsch et al., 2005); 4, B. caledonica (n57; Coenye et al.,
2001); 5, B. sediminicola HU2-65WT (Lim et al., 2008); 6, B.
xenovorans LB400T (Goris et al., 2004). Values are percentages of total
fatty acids.
Fatty acid
1
2
3
4
5
6
Summed feature*
8
41.1 35.6±2.1 44.3 34.2±1.7 31.3 27.3
3
18.5 13.6±1.9 17.6 14.5±1.8 17.0 19.1
2
5.8 8.1±1.1 5.6 7.4±0.9 8.0 8.5
16 : 0
14.9 14.7±0.9 13.8 13.6±1.6 20.8 18.2
16 : 0 3-OH
4.4 5.6±0.5 4.1 6.0±0.4 6.2 7.1
14 : 0
4.0 4.6±0.1 3.6 4.7±0.2 1.4 4.7
16 : 1 2-OH
2.6 3.5±0.7 2.3 2.7±0.4 1.3 2.2
17 : 0 cyclo
2.0 5.1±1.6 2.1 8.4±1.5 7.0 5.1
16 : 0 2-OH
1.7 3.6±0.5 2.1 2.4±0.4 2.2 2.2
2.0 2.5±0.7 1.8 3.7±0.7 2.9 3.6
19 : 0 cyclo v8c
18 : 1 2-OH
1.0 1.7±0.2 1.4 1.1±0.3 0.8 0.9
*Summed features represent groups of two or more fatty acids that
could not be separated under the conditions used. Summed feature 2
comprises 14 : 0 3-OH, 16 : 1 iso-I, an unidentified fatty acid of
equivalent chain-length 10.95 and 12 : 0 aldehyde; summed feature 3
comprises 16 : 1v6c and 16 : 1v7c; summed feature 8 comprises
18 : 1v6c and 18 : 1v7c.
Triplicate growth curves of strain PNG-AprilT in oxic R-2A
broth were used to determine the optima and tolerance
limits towards temperature (4–48 uC), pH (2–9), salt (0–
3.6 %), arsenate (AsV) and arsenite (AsIII). The nonmanipulated variables in each experiment were T532 uC,
pH 7, no NaCl, no AsV and no AsIII. Strain PNG-AprilT
showed growth between 4 uC and 40 uC, with a broad
optimum of 30–37 uC. In citrate-, MES-, Trizma- and PBSbuffered media, the isolate grew between pH 3.5 and
pH 8.3, showing fastest growth at pH 5–6, and stayed
viable at pH 3 for up to 5 weeks. Growth of its closest
relative, B. fungorum DSM 17061T, occurred between
pH 3.5 and pH 8.5 and was fastest at pH 6–7. Strain PNGAprilT showed salt tolerance of optimally 0–1.0 %, with a
maximum of 2.7 % NaCl. The novel strain was able to grow
in the presence of 0.5 mM AsV or AsIII, with the growth
rate slightly decreasing with increasing As concentration;
concentrations above 0.5 mM were not tested. When
grown in microoxic medium PNG-AR2, the isolate
reduced ~1.6 % of the AsV provided to AsIII, indicating
detoxifying rather than respiratory arsenate reduction. At
the isolation site in Tutum Bay, typical conditions in the
top 1 m of sediment were 32–41 uC, pH 6.1–6.2, 0.3–
4.7 mM AsV and ,0.1 mM AsIII (Price et al., 2007). Pore
waters represent a mixture between As-rich (13 mM)
http://ijs.sgmjournals.org
hydrothermal fluids of low salinity (~0.3 %) and local
seawater containing 3.0 % (w/v) NaCl (Pichler et al., 1999;
Price et al., 2007). The tolerance limits of strain PNGAprilT suggest its potential distribution across major parts
of Tutum Bay, except where T.40 uC.
Biochemically the novel strain was characterized in regards
to its C sources, electron donors, electron acceptors and
enzyme activities. Cultures of strain PNG-AprilT, B.
fungorum DSM 17061T, B. phytofirmans DSM 17436T, B.
caledonica DSM 17062T and B. sediminicola DSM 21400T
were tested for their utilization of carbohydrates. Colonies
grown on R-2A agar were suspended in API CHB medium
(bioMérieux), dispensed into API 50CH microtest galleries
(bioMérieux) and incubated at 32 uC, in order to identify
carbohydrates used in aerobic metabolism. Differences in
substrate utilization are summarized in Table 2. Carbohydrates used by all five species were D-glucose, D-fructose,
D-galactose, D-ribose, D-xylose, DL-arabinose, D-mannose,
L-rhamnose, DL-fucose, cellobiose, glycerol, D-adonitol,
inositol, D-mannitol, D-sorbitol and DL-arabitol.
Carbohydrates not used by any of the five species were
sucrose, melibiose, turanose, D-tagatose, L-sorbose, melezitose, raffinose, starch, glycogen, dulcitol, erythritol, Nacetylglucosamine and inulin. Strain PNG-AprilT was also
tested for growth in MSB (Stanier et al., 1966) and B-litho
media with individual addition of acetate, succinate,
citrate, malate and benzoic acid (20 mM each). The isolate
showed aerobic growth on all of these substrates, whereas
B-litho media amended with methanol or methylamine did
not support growth. Oxidation/fermentation tests (Hugh &
Leifson, 1953) showed oxidative metabolism of carbohydrates as specified in Table 2.
In anoxic R-2A broth under N2 headspace, strain PNGAprilT showed no growth. Purple broth tests for fermentation of glucose, lactose and sucrose were negative. As in all
known species of the genus Burkholderia, metabolism of
the novel isolate was obligately respiratory. Anoxic R-2A or
TS broth amended with nitrate (10 mM) supported
growth, albeit slower than aerobic growth. No growth
occurred with sulfate, thiosulfate, ferric iron or arsenate
offered as electron acceptors. Nitrate reduction by strain
PNG-AprilT was confirmed by a nitrate broth test with B.
fungorum DSM 17061T as a positive control and B.
sediminicola DSM 21400T as a negative control.
Lithoautotrophic metabolism was tested in oxic medium
B-litho with addition of nitrite, sulfide, thiosulfate, sulfite
or arsenite as electron donors (0.5 mM each). Weak
growth of strain PNG-AprilT was observed with thiosulfate
and nitrite, as confirmed by the formation of protein
(0.4–0.6 mg l21 and 0.1–0.3 mg l21, respectively). Lithoautotrophic growth on thiosulfate also occurred in the type
strains of B. fungorum, B. phytofirmans, B. caledonica and B.
sediminicola. Repeating the experiment in S-6 medium for
thiobacilli (ATCC medium 290, containing 40 mM
thiosulfate) confirmed these results. All thiosulfate-oxidizing cultures produced minor amounts of sulfate
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191
A. Rusch and others
Table 2. Phenotypic comparison of strain PNG-AprilT to type strains of closely related species of the genus Burkholderia
Strains: 1, PNG-AprilT; 2, B. fungorum DSM 17061T; 3, B. phytofirmans DSM 17436T; 4, B. caledonica DSM 17062T; 5, B. sediminicola DSM 21400T;
6, B. megapolitana LMG 23650T (data from Vandamme et al., 2007); 7, B. phenazinium LMG 2247T (Coenye et al., 2001); 8, B. xenovorans LB400T
(Goris et al., 2004); 9, B. rhynchosiae WSM3937T (De Meyer et al., 2013). Data for taxa 1–5 from this study except where indicated otherwise. +,
Positive; 2, negative; W, weak; S(+), substrate-dependent, ONPG positive; ND, not determined. O/F, oxidation/fermentation test.
Characteristic
Growth at/with:
37 uC
1.5 % (w/v) NaCl
3.0 % (w/v) NaCl
10 % (w/v) lactose
0.03 % (w/v) cetrimide
pH for growth
Range
Optimum
Nitrate reduction
Catalase
Oxidase
b-Galactosidase
O/F oxidation of:
D-Glucose
D-Fructose
D-Xylose
Maltose
Adonitol
Utilization of:
L-Xylose
Xylitol
Sucrose
Trehalose
Citrate
1
2
3
4
5
+
+
2
+
2
+a*
+a
2a
2a
+a
2b
+b
+b
+b
+b
2a
2a
2a
+a
2a
2c
+c
–c
3.5–8.3
5–6
+
+
+
+
3.5–8.5
6–7
+
+a
+a
2a
ND
ND
6–8c
ND
ND
ND
4–9
ND
ND
ND
ND
ND
ND
ND
2b
+b
2b
2a
+a
2a
2a
2
+c
+c
c
S(+)
2
2
2
2
2
2
+
2
2
+
+
2
+
+
+
+
+b
+b
+b
2b
2b
+a
+a
+a
+a
+a
2c
2c
2c
2c
2
2
2
+a
+a
+a
2a
2a
ND
ND
2
+
2
2
2
+
+
+
2
2
+
+
2
2
+
+
+
2
2
+
+
+
2
2
2b
2
+
2
+
2
2
2
2
2
+
ND
2
ND
ND
ND
ND
ND
ND
+
+
2
+
2
2
2
2
+
ND
W
W
ND
ND
ND
6
ND
2
2
2
2
ND
2
2
7
8
9
+
2
2
2
2
2
2
2
2
2
+
+
+
ND
ND
W
2
ND
ND
ND
ND
ND
+
*Data from: a, Coenye et al. (2001); b, Sessitsch et al. (2005); c, Lim et al. (2008).
(,35 mM in B-litho, 0.9–1.8 mM in S-6 medium). In
nitrite-oxidizing cultures of strain PNG-AprilT, mean
nitrate production was 41 mM, while 132 mM nitrite were
consumed. Growth with thiosulfate as an electron donor
has been described previously for mixotrophic cultures of
‘Burkholderia kururiensis subsp. thiooxydans’ (Anandham
et al., 2009) and a few Burkholderia strains isolated from
soil (Jung et al., 2005; Anandham et al., 2008) and acid
mine drainage (Bhowal & Chakraborty, 2011). Nitrite
oxidation has been demonstrated in one strain of
Burkholderia cepacia; an indirect pathway via nitric oxide
was suggested (Matsuzaka et al., 2003). Genes for the
catalysing enzymes occur in 15 species of the genus
Burkholderia [NCBI-RefSeq (http://www.ncbi.nlm.nih.gov/
refseq/), Sept. 2014]. Nitrite oxidation may also happen as
alternate reaction of certain catalases (Sakai et al., 2000).
Strain PNG-AprilT and the type strains of its four closest
relatives grew in N-free mineral salts medium amended with
mannitol (2 %), demonstrating their ability of N2-fixation.
Diazotrophy is widespread in the genus Burkholderia
(Coenye & Vandamme, 2003; Suárez-Moreno et al., 2012).
192
Applying well-established testing procedures (MacFaddin,
1980; Weiner & Penha, 1990; Leboffe & Pierce, 2011),
strain PNG-AprilT was characterized as possessing catalase,
oxidase, lipase/esterase and b-galactosidase. The enzymes
b-glucosidase, urease, lysine decarboxylase, tryptophanase,
DNase, gelatinase and amylase were not detected. A
comparison with closely related species of the genus
Burkholderia is provided in Table 2. Strain PNG-AprilT
differed from most of its nearest phylogenetic neighbours
in four or more traits, and as one of the few described
members of the genus Burkholderia that were not isolated
from soil or phytosphere, it is also distinct by habitat.
Pure cultures of strain PNG-AprilT, B. fungorum DSM
17061T and B. phytofirmans DSM 17436T were grown in
R-2A broth, before DNA was extracted and purified. DNA–
DNA hybridization at 68 uC was determined spectrophotometrically in duplicate (De Ley et al., 1970; Huss
et al., 1983) by the Identification Service of the LeibnizInstitut Deutsche Sammlung von Mikroorganismen und
Zellkulturen (DSMZ; Braunschweig, Germany). Strain
PNG-AprilT showed DNA–DNA relatedness duplicate
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Arsenic-tolerant facultatively lithotrophic Burkholderia
values of 35.0 % and 36.7 % with B. fungorum DSM 17061T,
and 10.3 % and 20.5 % with B. phytofirmans DSM 17436T.
Applying the recommended threshold value of 70 % DNA–
DNA hybridization (Wayne et al., 1987), strain PNG-AprilT
clearly belongs to a species different from its two closest
relatives within the genus Burkholderia. The DNA base
composition of the novel isolate was quantified by HPLC
analysis of enzymically hydrolysed DNA (Cashion et al.,
1977; Mesbah et al., 1989; Tamaoka & Komagata, 1984) and
was carried out by the Identification Service of the DSMZ.
The DNA G+C content of strain PNG-AprilT was
62.0 mol%, which is within the range of 61–69 mol%
typically observed for members of the genus Burkholderia.
L. Burcea, J. C. Minol and J. A. Armstrong assisted with growth media
preparation and general maintenance of our cultures.
On the basis of data obtained in this study, strain PNGAprilT represents a novel species of the genus Burkholderia,
for which the name Burkholderia insulsa sp. nov. is proposed.
subsp. thiooxydans subsp. nov., a facultative chemolithoautotrophic
thiosulfate oxidizing bacterium isolated from rhizosphere soil and
proposal for classification of the type strain of Burkholderia
kururiensis as Burkholderia kururiensis subsp. kururiensis subsp. nov.
Arch Microbiol 191, 885–894.
Description of Burkholderia insulsa sp. nov.
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Jee, H. J. (2009). Mixotrophic metabolism in Burkholderia kururiensis
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Burkholderia insulsa (in.sul9sa. L. fem. adj. insulsa unsalted,
bland, boring; in reference to the unsurprising characteristics of the type strain).
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Cells are motile, Gram-stain-negative, non-sporulating,
straight rods (2–3 mm long, 1 mm wide). Colonies on R-2A
agar are white to cream with a smooth edge. Grows at 4–
40 uC (optimally at 30–37 uC), at pH 3.5–8.3 (optimum
pH 5–6) and with salt concentrations of up to 2.7 % NaCl
(optimum 0–1.0 %). Tolerates at least 0.5 mM AsV or AsIII
and 10 % (w/v) lactose, but not cetrimide (0.03 %, w/v).
The strictly respiratory metabolism uses O2 and nitrate, but
not sulfate, thiosulfate, FeIII or AsV for electron acceptors.
Carbon sources include D-glucose, D-fructose, D-galactose,
D-ribose, DL-xylose, DL-arabinose, D-mannose, L-rhamnose,
DL-fucose, cellobiose, lactose, glycerol, xylitol, inositol, Dmannitol, D-sorbitol, DL-arabitol, succinate, malate, citrate,
acetate and benzoic acid, but not melibiose, sucrose,
trehalose, turanose, D-tagatose, methanol or methylamine.
Slow growth occurs lithoautotrophically with CO2 as sole C
source, O2 as electron acceptor and thiosulfate or nitrite as
electron donors; sulfite, sulfide or AsIII do not support
lithotrophic growth. N2 can be used as sole N source.
Bligh, E. G. & Dyer, W. J. (1959). A rapid method of total lipid
extraction and purification. Can J Biochem Physiol 37, 911–917.
Produces catalase, oxidase, lipase and b-galactosidase, but
not b-glucosidase, urease, amylase, gelatinase, DNase,
tryptophanase or lysine decarboxylase. The main respiratory quinones are ubiquinones Q-8 and Q-7.
Predominant cellular fatty acids are summed feature 8
(cis-vaccenic acid, cis-12-oleic acid), summed feature 3 (cis9- and cis-10-palmitoleic acid) and palmitic acid.
The type strain, PNG-AprilT (5DSM 28142T5LMG 28183T),
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