System. Appl. Microbiol.

27, 653–660 (2004)

http://www.elsevier.de/syapm

Mycobacterium fluoranthenivorans sp. nov.,

a Fluoranthene and Aflatoxin B1 Degrading Bacterium
from Contaminated Soil of a Former Coal Gas Plant
D. Hormisch1,4, I. Brost2, G.-W. Kohring1, F. Giffhorn1, R. M. Kroppenstedt3, E. Stackebrandt3, P. Färber2, and
W. H. Holzapfel2
1
Saarland University, Applied Microbiology, Saarbruecken, Germany
2
Federal Research Center for Nutrition and Food, Institute of Hygiene and Toxicology, Karlsruhe, Germany
3
DSMZ – Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Braunschweig, Germany
4
LUFA – Landwirtschaftliche Untersuchungs- und Forschungsanstalt, Speyer, Germany

Received: May 21, 2004

Summary
Mycobacterium strain FA4T was isolated with fluoranthene as the single carbon source from soil of a for-
mer coal gas plant, polluted with polycyclic aromatic hydrocarbons. The physiological properties, fatty
acid pattern, and the 16S ribosomal RNA gene sequence indicated membership to the genus Mycobac-
terium, but were different from all type strains of Mycobacterium species. Based on comparative 16S
rRNA gene sequence analyses strain FA4T could be assigned to the Mycobacterium neoaurum taxon
showing 98% sequence similarity to M. diernhoferi as its closest neighbour. The occurrence of epoxymy-
colate in the cell wall differentiates FA4 from all members of this taxon which synthesize wax-ester my-
colates in addition to alpha-mycolates. Strain FA4T is able to degrade aflatoxin B1. This biological at-
tribute might be useful in biological detoxification processes of foods and feeds. From the investigated
characteristics it is concluded that strain FA4T represents a new species, for which we propose the name
Mycobacterium fluoranthenivorans sp. nov. The type strain of Mycobacterium fluoranthenivorans is
FA4T (DSM 44556T = CIP 108203T).

Key words: Mycobacterium fluoranthenivorans sp. nov. – polycyclic aromatic hydrocarbons – fluoran-
thene – aflatoxin B1 – degradation

Introduction
Among toxic waste product degrading bacteria, nu-
merous strains from the genera Rhodococcus and My-
cobacterium have been isolated from the environment
and were described in detail [6, 7, 12, 13, 16, 27]. Some
of these organisms are able to degrade chlorinated com-
pounds like vinyl chloride [11] and pentachlorophenol
[9], or compounds from the chemically diverse group of
polycyclic aromatic hydrocarbons (PAH’s) [2, 5, 12, 28,
34, 36]. These bacterial strains can either utilise a wide
variety of xenobiotics as growth substrates or they are re-
stricted to only a few or a single hazardous compound.
Growth of Mycobacterium vanbaalenii [20], for example,
is described with naphthalene, phenanthrene, anthracene,
fluoranthene, pyrene, 1-nitropyrene, 3-methylcholan-
thene, 6-nitrocrysene and benzo[a]pyrene. Mycobacteri-
um frederiksbergense [41] mineralises phenanthrene, flu-
oranthene or pyrene and Mycobacterium hodleri [21]
grows with fluoranthene as the only carbon source, but
co-oxidizes other PAH’s during growth with fluoran-
thene.
Aflatoxins with the naturally occurring subgroups B1,
B2, G1, G2, M1 and M2 are bisfurano-coumarin deriva-
tives. They are structurally closely related to the above
mentioned chemical compounds. Aflatoxins are pro-
duced during the secondary metabolism of filamentous
fungi. Especially aflatoxin B1 (AFB1) belongs to the most
dangerous naturally occurring mycotoxins because of its
hepatotoxic, hepatocarcinogenic, mutagenic and terato-
genic effects, both on humans and on animals. AFB1 is
produced both under moderate and under subtropic and
tropic climatic conditions. Because of its hazardous nature,
Genbank accession number: AJ617741

0723-2020/04/27/06-653 $ 30.00/0
654 D. Hormisch et al.
different physical and physico-chemical techniques have
been developed to detoxify and to degrade AFB1 [40].
AFB1 is a physically and physico-chemically stable
molecule, e.g. temperatures > 250 °C are necessary for ef-
fective destruction. Foods treated for physical degrada-
tion of AFB1 are not acceptable for human consumption.
Therefore, biological degradation of AFB1 without sig-
nificant alteration of the sensory characteristics of a
food, would be most advantageous, particularly in situa-
tions of food scarcity. During the search for bacterial de-
graders of AFB1, strains from different habitats, includ-
ing soils from a former coal gas plant, were screened for
their ability to degrade AFB1. Among the few positive
strains, one was found to belong to the genus Mycobac-
terium but could not be allocated to any known species
of this genus, thus representing a putative new species.
This paper describes Mycobacterium strain FA4T which
is able to degrade AFB1 most effectively.
Material and Methods
Isolation
PAH polluted soils from a former coal gas plant in Saar-
bruecken-Burbach, Germany, were screened for potential PAH
degraders; in this process, strain FA4T was isolated by the fol-
lowing procedure: The sampling comprised 3 g of soil which
were added to 25 ml of liquid medium containing in 1000 ml
H2O: 0.9 g KH2PO4, 2.5 g Na2HPO4· H2O, 0.2 g MgCl2· H2O,
0.5 g KNO3, 0.1 g (NH4)2SO4, 0.2 g NH4Cl, 3.59 mg
CaCl2· H2O, 1.66 mg FeCl3· H2O, 10 ml SL4 [30], and 10 ml of
vitamin solution [31], adjusted to a final pH of 7.2. As a carbon
source 2.5 ml of fluoranthene (10 mg/ml) dissolved in hep-
tamethylnonane (ICN Biochemicals) were loaded on top of the
medium, which provided the organisms with a constant concen-
tration of fluoranthene in the water phase, determined by the
partition coefficient. The cultures were incubated at 28 °C on a
rotary shaker. After visible growth, the enrichment cultures were
transferred to fresh media for at least three times and then sepa-
rated on R2A agar plates (Difco Lab.). Single colonies were
grown again in the described liquid medium and finally tested
for purity on R2A agar plates.
Characterisation and biochemical tests
All tests followed methods indicated [41] and were per-
formed in duplicate. As a control, all tests were also carried out
with M. hodleri which exhibited the same results as reported in
the literature [21]. Morphological analysis, Gram staining and
acid-alcohol fastness were investigated by phase contrast and
bright field microscopy. The ability to grow at different temper-
atures, pigment production and photoreactivity were deter-
mined after 2 weeks of growth in Middelbrook medium [23].
Tests for urease [10], catalase [22] and nitrate reductase activity
or Tween 80 hydrolysis [1] were performed as described in the
citations. Growth investigations with sugars and sugar alcohols
[40], and other physiological tests were performed as described
[19, 41].
Growth with different PAH’s as single carbon sources was in-
vestigated in the same medium described under “isolation”, sup-
plemented with 10% (w/v) of each PAH dissolved in hep-
tamethylnonane (10 mg/ml). Degradation of the PAH’s was de-
termined by reverse phase HPLC (Nucleosil 120-5-C-18 col-
umn, 48.5% methanol and 3% acetic acid in water as the mo-
bile phase, UV detection, Beckman System Gold) after diluting
samples of the heptamethylnonane phase 1:350 with methanol
prior to injection.
Analytical methods
As described [37], 40 mg wet weight of cells were saponified,
methylated and extracted. The derived fatty acid methyl esters
(FAMEs) were analysed by GC (model 5898A, Hewlett Packard,
controlled by MIS software, Microbial ID) and the “Microbial
Identification” standard software package [35]. Thin layer chro-
matography investigation of mycolic acids followed described
methods [24, 26]. Mycolic acids were extracted from saponified
cells and transferred to their bromophenacyl esters [3, 25]. After
addition of low and high molecular mass standards (Ribi Im-
munoChem Research) the mixture was separated and analysed
by HPLC (C18 Ultrasphere XL cartridge column, 35 °C,
Hewlett Packard Series 1050 operated with the Sherlock System
software, MIDI Inc.). For the analysis of the mycolic acid cleav-
age products (MACP) the cells were treated as described above
for fatty acids. To enhance the pyrolysis of the mycolic acids 10
µl of 0.2 M methanolic TMSH was added to the fatty acid/my-
colic acid methyl esters extract and analysed by GC [29].
16S rRNA gene sequence determination
Extraction of the DNA, PCR-mediated amplification of the
16S rDNA and purification of the PCR products were done as
described [33]. The purified amplificates were analysed with an
Applied Biosystems 373A DNA Sequencer. The 16S rRNA gene
sequence was aligned manually with published sequences from
representatives of the actinomycete sublines of descent included
in the DSMZ database of sequences. Neighbour joining analysis
and calculation of bootstrap values were done according to the
PHYLIP program [8].
AFB1 degradation experiments
The method for the quantitative determination of AFB1 is
based on the CEN/DIN method prEN 14123 (“Foodstuffs – De-
termination of aflatoxin B1 and the sum of aflatoxin B1, B2, G1
and G2 in peanuts, pistachios, figs and paprika powder”). Sim-
plification was possible by using a miniaturised in vitro test sys-
tem, whereby there was no need for the use of an immuno-affin-
ity column clean-up of the test samples. In contrast to complex
foodstuff like peanuts, pistachios or maize, which are rich in
protein and fat/oil, the complete medium used in these tests did
not require a clean-up of samples. The test cultures of strain FA4
have been regularly subcultured on an AFB1-containing medium
at least three times for 12 hours at 30 °C for a possible induc-
tion of enzymes involved in AFB1 degradation. Media used for
the precultivation of the cells and used in the degradation exper-
iments itself, were supplemented with AFB1 to an initial concen-
tration of 2.5 ppm. The degradation experiments were per-
formed on a rotary shaker at 30 °C in the dark. Samples for
analysis of the AFB1 concentration were withdrawn at 12 h, 36
h and 72 h after inoculation. The remaining AFB1 concentration
was quantitatively determined by HPLC (stationary phase:
LiChroCart 250-4 Hypersil ODS [5 µm] [MERCK, Germany];
mobile phase: CH3CN:CH3O:H2O [25: 25 :50/vol:vol:vol];
detection: DAD at 365 nm).
Results and Discussion
Morphological and physiological properties
The organism FA4T was rod shaped (2 µm length,
1 µm diameter), Gram-positive, acid-alcohol-fast and not
motile. The strain was nonchromogenic and grew at
 Mycobacterium fluoranthenivorans sp. nov. 655

Table 1. Physiological properties of M. fluoranthenivorans DSM 44556T, M. diernhoferi, M. hodleri and M. vanbaalenii.

Mycobacterium Mycobacterium Mycobacterium Mycobacterium

diernhoferi d fluoranthenivorans hodleri d vanbaalenii e

Formation of pigment a N N S S
Gram stain + + + +
Acid fast stain + + + +
Growth at 20 °C/28 °C/37 °C + + + +
Growth at 42 °C – – – –
Tween 80 hydrolysis – – – (+) c +
Nitrate reduction + – – +
Urease activity n.a. – +c +
Catalase activity – + + +
Growth on:
Xylose + + + –
Trehalose – + + (–) c –
Sorbitol – + + –
Utilisation b of:
N-acetyl-d-glucoseamine – + + n.a.
D-Glucosaminic acid – + – n.a.
Gluconate + + + n.a.
D-Sucrose – + – –
D-Turanose – + – n.a.
L-Rhamnose – + + –
D-Ribose – + + n.a.
D-Arabitol – + + n.a.
I-Inositol + + + n.a.
Mannitol + + + n.a.
Citrate – + – –
2-Oxoglutarate + + + n.a.
Glutarate n.a. + – n.a.
Succinate – + + n.a.
L-Alanine + + + n.a.
L-Aspartate – + + n.a.
L-Leucine + + + n.a.
L-Proline + + + n.a.
L-Valine – + + n.a.
Putrescine + + + n.a.
Acetamide – + – n.a.
Phenyl acetic acid – + – n.a.
Benzoate – + – n.a.
4-Amino-benzoate – + – n.a.
4-Hydroxy-benzoate – + – n.a.
Quinate – + + n.a.
P-nitrophenylphosphoryl-choline + + – n.a.
a
S, scotochromogenic, N, nonchromogenic.
b
Assimilation of auxanographic substrates was detected photometrically by means of reduction of the redox dye MTT (Kirchhof
et al. 1992). + = E540(test) – E540 (control) > 0.129; – = E540 (test) – E540 (control) <0.129.
c
Data from Kleespies et al. [21], d Data from Willumsen et al. [41], e Data from Khan et al. [20] n.a. data not available

20 °C, 28 °C and 37 °C but not at 42 °C. In contrast to a
well expressed catalase activity, no urease or nitrate re-
duction activity was observed, and Tween 80 was not hy-
drolysed (Table 1).
From the group of polycyclic aromatic hydrocarbons,
only the isolation substrate fluoranthene was metabolised
by strain FA4T. No growth was detected with naphtha-
lene, anthracene, phenanthrene, acenaphthene, acenaph-
thylene, fluorene, chrysene, or pyrene as the single carbon
source (Table 2), and the strain could not utilise biphenyl.
However, phenylacetic acid, benzoate, benzoate deriva-
tives and p-nitrophenylphosphoryl-choline were utilised
as aromatic substrates (Table 1).
Table 1 summarises the physiological characteristics of
strain FA4T, including its utilisation of a broad spectrum
of carbon sources such as sugars, sugar alcohols, amino
acids, organic acids and mono aromatic compounds.
Degradation of AFB1
AFB1 degradation by Mycobacterium strain FA4 was
relatively rapid and effective, leaving no detectable AFB1
656 D. Hormisch et al.

Table 2. Growth of M. fluoranthenivorans DSM 44556 T, M. hodleri, M. vanbaalenii and M. frederiksbergense with biphenyl or sev-
eral polycyclic aromatic hydrocarbons as single carbon sources.

Mycobacterium Mycobacterium Mycobacterium Mycobacterium

fluoranthenivorans hodleri a vanbaalenii b frederiksbergense c

Biphenyl – n.a. + n.a.

Naphthalene – n.a. + n.a.
Anthracene – – + –
Phenanthrene – – + +
Acenaphthene – n.a. n.a. n.a.
Acenaphthylene – n.a. n.a. n.a.
Fluorene – – n.a. –
Fluoranthene + + + +
Chrysene – n.a. n.a. n.a.
Pyrene – – + +
a
Data from Kleespies et al. [21], b Data from Khan et al. [20], c Data from Willumsen et al. [41] n.a. data not available

Table 3. Composition of FAMEs derived from whole-cell hydrolysates of strain FA4T and members of the Mycobacterium neoaurum
cluster.

Fatty acid (%) FA4T DSM 43524T DSM 44346T DSM 44077T DSM 44183T DSM 44074T

10:0 0.66 – – – – –
12:0 0.60 – – – – –
14:0 10.02 6.23 5.45 4.73 5.74 5.72
15:0 – – 0.66 0.35 1.05 –
16:1 cis-7 – 2.17 6.30 0.54 1.49 3.48
16:1cis-9 13.46 0.77 – 0.83 2.42 5.23
16:1cis-10 4.60 5.65 3.41 3.56 6.07 –
16:0 42.11 25.27 31.44 26.87 25.17 28.31
16:0 10-methyl 2.17 – – – 0.76 –
17:0 – – – 0.42 – –
18:2 – – – 0.45 0.63 –
18:1cis-9 16.80 20.57 34.83 14.55 20.27 30.46
18:0 3.37 3.98 2.27 3.00 1.24 1.96
18:0 10-methyl 5.09 11.15 1.36 9.33 18.46 5.57
20:0 1.11 – 1.36 0.46 – –
18:0 alcohol – 18.00 10.78 16.77 13.93 15.07
20:0 alcohol – 4.57 1.71 16.32 2.77 1.74

Values are percentages of total fatty acids. Secondary alcohols that are released from wax esters are included. Examples of abbrevia-
tions: 16:0, hexadecanoic acid (palmitic acid);
18:1 cis-9, cis-9-octadecenoic acid (oleic acid); 18:0 10 methyl, 10-methyloctadecanoic acid (tuberculostearic acid); 20: 0 alcohol, 2-
eicosanol.
M. chitae DSM 43633, M. farcinogenes DSM 43637, M. fortuitum DSM 46621, M. peregrinum DSM 43271, M. porcinum DSM
44242, M. senegalensis DSM 43656, M. smegmatis DSM 43756.

after 72 h. The AFB1 concentration was reduced to
amounts of 70% to 80% of the initial concentration
within 36 hours. In further experiments, cell-free extracts
of Mycobacterium FA4 have been used in the same way,
both to confirm these observations and the enzymatic na-
ture of the degradation activity (data not shown).
Chemotaxonomic properties
The analysis of the fatty acids revealed a pattern com-
posed of straight chain saturated and unsaturated fatty
acids with an even number of carbon atoms. The diag-
nostic 10-methyl branched fatty acids were found in ad-
dition (Table 3). The pyrolysis esters of the mycolic acids
showed a chain length of 22, 24 and 26 carbon atoms
(Table 4). The mycolic acid pattern separated by TLC
was composed of alpha-mycolates and epoxy mycolates
(Table 4). Analyses of mycolic acids by HPLC revealed a
characteristic UV-HPLC chromatogram with a two-peak
cluster that emerged late and close together after 6–8
minutes (Fig. 1). It seems that there is a correlation be-
tween this HPLC-chromatogram and the occurrence of
alpha-mycolate and epoxy-mycolates because this two-
peak cluster is always present in mycobacteria species
which synthesise alpha-mycolates and epoxy-mycolates
i.e. M. agri, M. chelonae, M. chitae, M. farcinogenes,
 Mycobacterium fluoranthenivorans sp. nov. 657

Table 4. Mycolic acids and pyrolysis esters of strain FA4T and members of the M. neoaurum group a.

Mycolic acidsa FA4T DSM 43524T DSM 44346T DSM 44077T DSM 44183T DSM 44074T

alpha-mycolates + + + + + +
alpha′-mycolates (+) – – – – –
Methoxy-mycolates – – – – – –
Keto-mycolates – + + + + +
Epoxy-mycolates + – – – – –
Waxester-mycolates – + + + + +
w-1-Methoxy-mycolates – – – – – –
Pyrolysis esters
C22: 0 + + + n.d. + +
C24: 0 + + – n.d. + –
C26: 0 + – – n.d. – –
a
Data from Willumsen et al., 2001; b +, present; –, absent; (+), small amounts present; n.d., no data.
M. diernhoferi DSM 43524T, M. frederiksbergense DSM 44346 T, M. gadium DSM 44077 T, M. hodleri DSM 44183T, M. neoaurum
DSM 44074T.

Fig. 1. Mycolic acid HPLC

elution profile of FA4T
(= DSM 44556T). LIS, low-
molecular-mass internal stan-
dard; HIS, high-molecular-
mass internal standard.

members of the M. fortuitum., M. goodie, M. pere-
grinum, M. smegmatis, M. wolinskyi, M. porcinum and
M. senegalense [4, 15].
Phylogenetic analysis
Determination of the 16S rRNA gene sequence and the
comparison with other sequences of members of the
genus Mycobacterium revealed that strain FA4T belongs
to M. neoaurum subgroup of fast-growing mycobacteria.
The closest relationship obtained with the type of a de-
scribed species was M. diernhoferi ATCC 25795T (98%
sequence similarity). Strain ATCC BAA-823, proposed as
“Mycobacterium hackensackense” [17] (AY266138)
shows identity with the homologous sequence of strain
FA4T.
Differentiation of strain FA4T from other mycobacteria
Strain FA4T could clearly be separated from the other
members of the M. neoaurum complex by its fatty acid
pattern. FA4T lacks the two secondary alcohols C18:0
658 D. Hormisch et al.

Table 5. Mycolic acids of Mycobacterium sp. FA4T and other species showing alpha- and epoxy-mycolates in their mycolic acid pat-
terns.

DSM 43633T

DSM 43637T

DSM 46621T

DSM 43271T

DSM 44242T

DSM 43656T

DSM 43756T
Mycolic acids a

FA4T
alpha-mycolates + + + + + + + +
alpha′-mycolates (+) + (+) + (+) + (+) +
methoxy-mycolates – – – – – – – –
keto-mycolates – – – – – – – –
epoxy-mycolates + + + + + + + +
waxester-mycolates – – – – – – – –
omega-1-methoxy-mycolates – – – (+) (+) (+) (+) –
a
Data from Vincent Levy-Frebault & Portales, 1992; Hinrikson & Pfyffer, 1994.
M. chitae DSM 43633T, M. farcinogenes DSM 43637 T, M. fortuitum DSM 46621T, M. peregrinum DSM 43271T,
M. porcinum DSM 44242 T, M. senegalensis DSM 43656 T, M. smegmatis DSM 43756 T.

Fig. 3. AFB1 degradation by viable cells of Nocardia corynebac-

terioides DSM 44601 (former Flavobacterium aurantiacum,
DSM 12676) and Nocardia corynebacterioides DSM 20151 in
comparison to Mycobacterium fluoranthenivorans DSM 44556T
(FA4T). All strains were cultivated in Standard I medium con-
taining 2.5 ppm AFB1, at 30 °C under shaking conditions for
12 h, 36 h and 72 h.

Fig. 2. Dendrogram of 16S rRNA gene sequence relatedness

showing the phylogenetic position of strain DSM 44556 T within
the radiation of some fast growing species of the genus My-
cobacterium. Numbers at branching points refer to bootstrap
values (500 re-samplings). Scale bar, 5 inferred nucleotide sub-
stitutions per 100 nucleotides.
and C20:0 which are always present in members of the
M. neoaurum complex. This taxon includes M. diern-
hoferi, M. frederiksbergense, M. gadium, M. hodleri and
M. neoaurum [32, 41]. Instead of the secondary alcohols
a C26:0 mycolic acid pyrolysis ester is found in the ex-
tract of FA4T. This ester is missing in the other species of
this taxon (Table 3). FA4T synthesized alpha-mycolates
and epoxymycolates. This mycolic acid pattern is shared
by M. chitae, M. farcinogenes, M. fortuitum, M. pere-
grinum, M. porcinum, M. senegalense and M. smegmatis
[15] (Table 5) which are not related to FA4T by 16S rRNA
gene sequences. In members of the M. neoaurum complex
waxester mycolates instead of epoxy-mycolates are found
in addition to alpha-mycolates which are present in all
mycobacteria [15, 41].
In combination, the results of this investigation
demonstrate that the described strain FA4T represents a
new species of the genus Mycobacterium. Compared to
other PAH metabolising mycobacteria, there are pro-
found differences to bacteria with a broad PAH substrate
spectrum such as M. frederiksbergense or M. vanbaalenii
(Table 2). M. hodleri, which could also only use fluoran-
thene, exhibited many differences in the physiological
data as shown in Table 1. In contrast to M. fluoran-
thenivorans sp. nov., M. hodleri was scotochromogenic,
expressed urease activity and could not utilise most of the
mono aromatic substrates. The strain with the highest
similarity with respect to 16S rRNA gene sequence analy-
sis, M. diernhoferi, expressed also significant differences
in the utilisation of sugars, some amino acids and the
mono aromatic compounds (Table 1).

Phylogenetic and chemotaxonomic data strongly sup-
ported the description of a new species, because the simi-
larity to M. diernhoferi is only 98%. This value is signifi-
cantly below 99.0%, the borderline of species differentia-
tion in the genus Mycobacterium which could be shown
in the recent description of new Mycobacterium species.
The publication of Hong et al. [17] gives no indication
that the proposed species “Mycobacterium hackensack-
ense” will be validly described. Though similarities in the
mycolic acid patterns are obvious, identical 16S rRNA
gene sequence similarities do not exclude affiliation to
different species. Also, while strain FA4T was isolated
from a contaminated soil sample, strain ATCC BAA-823
originated from a 6-year old female patient with relapsed
pre-B-cell acute lymphocytic leukaemia.
Description of Mycobacterium fluoranthenivorans
sp. nov.
Mycobacterium fluoranthenivorans (flu.or.an.the.ni-
vo´rans, N.L. n. fluoranthene; connecting vowel, -i-; L.v.
vorare; to devour; L. part. adj vorans, devouring, digest-
ing; N.L. part. adj. fluoranthene, digesting fluoranthene).
Gram-positive, acid-alcohol-fast, non motile rod shaped
(length 2 µm, diameter 1 µm) bacterium. It is nonchro-
mogenic, catalase positive and grows between 20 °C and
37 °C. The strain utilises xylose, trehalose, sorbitol, N-
acetyl-D-glucoseamine, D-glucosaminic acid, gluconate,
D-sucrose, D-turanose, L-rhamnose, D-ribose, D-ara-
bitol, I-inositol, mannitol, citrate, 2-oxoglutarate, glu-
tarate, succinate, L-alanine, L-aspartate, L-leucine, L-pro-
line, L-valine, putrescine, acetamide, phenyl acetic acid,
benzoate, 4-amino-benzoate, 4-hydroxy-benzoate,
quinate, p-nitrophenyl-phosphoryl-choline. Mineralises
fluoranthene. The fatty acid pattern from whole-cell
methanolysates is composed of tetredecanoic acid (10%),
cis-9-hexadecenoic acid (14%), cis-10-hexadecenoic acid
(5%), hexadecanoic acid (42%), 10-methyl-hexadecanoic
acid, cis-9-octadecenoic acid (17%), octadecanoic acid
(3%), 10-methyl-octadecanoic acid (5%) and eicosanoic
acid (1%). TLC of mycolic acid methanolysates reveals
alpha-mycolates and epoxy-mycolates. The mycolic acid
UV-HPLC elution profile shows two-peak clusters that
emerge late and close together.
Isolated from PAH contaminated soils from the site of
a former coal gas plant in Saarbruecken-Burbach, Ger-
many.
The type strain is FA4T (= DSM 44556T = CIP 108203).
Acknowledgements
The support by (1) the EU Commission within the INCO-DC
project “Biological degradation of aflatoxins in fermented maize
and sorghum products” (contract no. ERBIC18 CT980315),
and (2) a grant of the Saarberg AG, Saarbruecken Germany, is
gratefully acknowledged. Views expressed in this paper do not
necessarily reflect those of the European Commission. The skill-
ful technical assistance by Birgit Hasper, Gabriele Pötter, Jolan-
tha Swiderski and Ina Kramer is gratefully acknowledged.
Mycobacterium fluoranthenivorans sp. nov. 659
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Corresponding author:
W. H. Holzapfel, Federal Research Center for Nutrition and Food,
Institute of Hygiene and Toxicology, Haid-und-Neu-Str. 9,
76131 Karlsruhe, Germany
Tel.: ++(0)721-6625-450; Fax: ++(0)721-6625-453;
e-mail: wilhelm.holzapfel@bfe.uni-karlsruhe.de