International Journal of Systematic and Evolutionary Microbiology (2004), 54, 1469–1476
DOI 10.1099/ijs.0.02873-0
Akkermansia muciniphila gen. nov., sp. nov.,
a human intestinal mucin-degrading bacterium
Muriel Derrien, Elaine E. Vaughan, Caroline M. Plugge
and Willem M. de Vos
Correspondence
Muriel Derrien
Laboratory of Microbiology, Wageningen University, Hesselink van Suchtelenweg 4, 6703 CT
Wageningen, The Netherlands
muriel.derrien@wur.nl
The diversity of mucin-degrading bacteria in the human intestine was investigated by combining
culture and 16S rRNA-dependent approaches. A dominant bacterium, strain MucT, was isolated
by dilution to extinction of faeces in anaerobic medium containing gastric mucin as the sole carbon
and nitrogen source. A pure culture was obtained using the anaerobic soft agar technique.
Strain MucT was a Gram-negative, strictly anaerobic, non-motile, non-spore-forming, oval-shaped
bacterium that could grow singly and in pairs. When grown on mucin medium, cells produced a
capsule and were found to aggregate. Strain MucT could grow on a limited number of sugars,
including N-acetylglucosamine, N-acetylgalactosamine and glucose, but only when a protein
source was provided and with a lower growth rate and final density than on mucin. The G+C
content of DNA from strain MucT was 47?6 mol%. 16S rRNA gene sequence analysis revealed
that the isolate was part of the division Verrucomicrobia. The closest described relative of strain
MucT was Verrucomicrobium spinosum (92 % sequence similarity). Remarkably, the 16S rRNA
gene sequence of strain MucT showed 99 % similarity to three uncultured colonic bacteria.
According to the data obtained in this work, strain MucT represents a novel bacterium belonging
to a new genus in subdivision 1 of the Verrucomicrobia; the name Akkermansia muciniphila
gen. nov., sp. nov. is proposed; the type strain is MucT (=ATCC BAA-835T=CIP 107961T).
The human gastrointestinal (GI) tract harbours diverse and
abundant microbiota, which have effects on the health and
disease of the host due to their close association (Hooper
& Gordon, 2001). A mucus layer covers the GI tract, providing a protective barrier for the underlying epithelium
against pathogenic micro-organisms, as well as chemical,
physical or enzymic damage. Mucus is a viscous gel mainly
composed of high-molecular-mass glycoproteins, termed
mucins. Mucins are composed of a peptide core rich in
serine and threonine residues that is decorated by oligosaccharides linked via O- or N-glycosidic bonds. The
oligosaccharides are composed of one or more of four
primary sugars (N-acetylglucosamine, N-acetylgalactosamine,
galactose and fucose) and are terminated by sialic acids
or sulfate groups. This mucus layer is considered to be an
ecological niche for intestinal microbiota. However, the
association of microbiota with the mucus is not well
understood. Mucus can serve as a barrier to protect the
Published online ahead of print on 14 May 2004 as DOI 10.1099/
ijs.0.02873-0.
Abbreviations: DGGE, denaturing gradient gel electrophoresis; GI,
gastrointestinal; MPN, most probable number.
The GenBank/EMBL/DDBJ accession number for the 16S rRNA gene
sequence of Akkermansia muciniphila MucT is AY271254.
02873 G 2004 IUMS
Printed in Great Britain
underlying epithelium from the attachment of pathogens
and also as a source of nutrients for commensal bacteria.
Degradation of mucin is regarded as a pathogenicity factor
since loss of the protective mucus layer may expose GI
tract cells to pathogens (Ruseler-van Embden et al., 1995;
Zhou et al., 2001). However, mucin also constitutes a carbon
and energy source for intestinal microbiota. It has been
estimated that 1 % of colonic microbiota is able to degrade
host mucin using enzymes (e.g. glycosidases and sulfatases)
that can degrade the oligosaccharide chains (Hoskins &
Boulding, 1981). Despite the apparent low level of mucindegrading bacteria, these species provide nutrients for other
resident bacteria, which can use the monosaccharides or
amino acids released from mucin degradation. Based on
their capacity to grow on mucin-containing media, isolates
belonging to the genera Ruminococcus, Bacteroides, Bifidobacterium and Clostridium have been shown to degrade
mucin (Salyers et al., 1977). By measuring the release of
reducing sugar monomers from the mucin polymer, it was
observed that only mixed cultures of faecal bacteria were
able to degrade mucin by more than 90 %, whereas pure
cultures of Bacteroides fragilis, Bifidobacterium longum and
Clostridium perfringens showed only partial degradation
(Willis et al., 1996). It is therefore likely that, in vivo, a cooperative process is required to achieve efficient degradation
of the complex structure of mucin.
1469
M. Derrien and others
The introduction of high-resolution molecular techniques
has improved analyses of complex microbial ecosystems.
The most important advance has been the use of the 16S
rRNA gene as a molecular fingerprint to analyse microbial
diversity. Molecular approaches have indicated that a
lack of knowledge regarding cultivation conditions has
hampered our view of the intestinal microbiota (Vaughan
et al., 2000). As a consequence, a substantial proportion of
the microbiota has not yet been cultured or described
(Zoetendal et al., 1998; Suau et al., 1999); this may be due
mainly to the lack of appropriate cultivation techniques.
However, new, alternative and improved cultivation
approaches are continuously being developed and recently
a number of novel species and genera have been cultured
from the GI tract: Roseburia intestinalis (Duncan et al.,
2002), Campylobacter hominis (Lawson et al., 2001),
Ruminococcus luti (Simmering et al., 2002), Anaerostipes
caccae (Schwiertz et al., 2002), Dorea longicatena (Taras et al.,
2002) and Victivallis vadensis (Zoetendal et al., 2003).
In the present study, mucin-degrading bacteria from
human faeces were enriched using a most probable
number (MPN) approach in which the medium contained
mucin as the sole carbon and energy source. The enrichments were analysed by denaturing gradient gel electrophoresis (DGGE) of PCR-amplified 16S rRNA gene
sequences. A single DGGE type dominated all the positive
MPN enrichments. The organism corresponding to the
dominant DGGE type was isolated and characterized; it
represents a novel intestinal bacterium, strain MucT, that
is able to use gastric mucin in pure culture.
A faecal sample from a healthy adult volunteer was freshly
collected in a polyethylene bag and 0?5 g was diluted into
9 ml sterile anaerobic Ringer’s solution containing 0?5 g
cysteine l21. This suspension was thoroughly mixed and
serially diluted (10-fold) in Ringer’s. Each dilution (1 ml)
was inoculated in triplicate into 9 ml bicarbonate-buffered
medium. This basal medium contained (l21): 0?4 g KH2PO4;
0?53 g Na2HPO4; 0?3 g NH4Cl; 0?3 g NaCl; 0?1 g
MgCl2.6H2O; 0?11 g CaCl2; 1 ml alkaline trace element
solution; 1 ml acid trace element solution; 1 ml vitamin
solution; 0?5 mg resazurin; 4 g NaHCO3; 0?25 g
Na2S.7–9H2O. The trace element and vitamin solutions
were as described previously (Stams et al., 1993). All compounds were autoclaved, except the vitamins, which were
filter-sterilized. This basal medium was supplemented with
0?7 % (v/v) clarified, sterile rumen fluid and 0?25 % (v/v)
commercial hog gastric mucin (Type III; Sigma), purified
by ethanol precipitation as described previously (Miller &
Hoskins, 1981). This medium is further referred to as
mucin medium. Unless indicated, incubations were done
in serum bottles sealed with butyl-rubber stoppers at 37 uC
under anaerobic conditions provided by a gas phase of
182 kPa (1?8 atm) N2/CO2 (80 : 20, v/v). Enrichments were
done in 30 ml serum bottles with 10 ml liquid volume.
Negative controls comprised one series of mucin media
that was not inoculated and another series that was
1470
inoculated, but not supplemented with mucin. Mucindegrading bacteria were quantified using the MPN
technique (n=3). The soft agar technique was used to
isolate a pure culture as follows: the highest dilution where
growth was observed was serially diluted in phosphate
buffer (pH 7) until 1029 dilution and the 1026 to 1029
dilutions were re-inoculated into the same medium
containing 0?75 % agar (agar noble; Difco). Single colonies
were picked, grown in mucin medium and re-inoculated
in soft agar mucin medium. This step was repeated until
purity.
Generation times were determined in mucin medium and
growth was analysed in triplicate by measuring absorbance
at 600 nm. The optimum pH and temperature were measured in triplicate on brain–heart infusion (BHI; Difco)
supplemented with 1 mM Na2S. Temperatures tested were
4–45 uC, at intervals of 5 uC; growth was determined at
pH 5–9, at intervals of 0?5 pH units (adjusted with HCl or
NaOH) at 37 uC. Cultures were incubated for at least
1 month.
Potential substrates for growth were tested at a concentration of 10 mM in the same liquid basal medium or in basal
medium supplemented with peptone, tryptone, casitone
and yeast extract at a concentration of 0?5 or 2 g l21.
Cultures were incubated for up to 4 weeks. Human gastric
mucin was isolated from HT-29 MTX human intestinal
cell lines and this mucin was added to the basal liquid
medium at a concentration of 0?05 %. Rich media BHI
and Columbia broth (Difco) and 16 g Wilkens–Chalgren
broth (WC broth; Oxoid) l21 were also tested as growth
substrates. To test the origin of the nitrogen source, the
solution containing NH4Cl was not added to the mucin
medium.
Cell morphology, motility and spore formation were
investigated using phase-contrast microscopy. The Gram
reaction was assessed using Gram staining as described
previously (Plugge et al., 2000). To test for the presence
of a capsule, an Indian ink suspension was used.
For TEM of strain MucT, cells were fixed with 0?25 %
glutaraldehyde. Negative staining was performed on 400
copper mesh grids with glow-discharged parladion carbonsupport film. Micrographs were recorded at a magnification of 40 0006 on a JEOL 1010 electron microscope
operating at 80 kV. For SEM, droplets of strain MucT were
put onto poly-L-lysine-coated Nucleopore polycarbonate
membranes (Costar). These membranes were fixed for 1 h
in 4 % glutaraldehyde in growing medium. Specimens were
dehydrated in a graded series of ethanol and critical-point
dried with carbon dioxide. The samples were glued onto a
sample holder using carbon adhesive tabs. Samples were
sputter-coated with 10 nm platinum in a dedicated preparation chamber (CT 1500 HF) and analysed with a field
emission SEM (JEOL 6300 F) at 5 kV.
The G+C content of DNA of strain MucT was determined
International Journal of Systematic and Evolutionary Microbiology 54
Akkermansia muciniphila gen. nov., sp. nov.
at the DSMZ (Deutsche Sammlung von Mikroorganismen
und Zellkulturen, Braunschweig, Germany) by HPLC
(Mesbah et al., 1989).
1
2
3
4
5
6
7
8
9
10
To monitor the dynamics of the human faecal mucindegrading population, DGGE analysis of 16S rRNA gene
amplicons was performed. DNA was extracted from the
faecal sample, enrichment cultures and a pure culture
isolated from the highest dilution. DNA isolation and the
amplification of the V6 to V8 regions of the 16S rRNA
gene from these samples were performed as described
previously (Zoetendal et al., 1998). PCR fragments were
separated by DGGE consisting of 8 % (v/v) polyacrylamide
(ratio of acrylamide to bisacrylamide, 37?5 : 1) and 0?56
Tris/acetate/EDTA (pH 8?0) (TAE) buffer; 100 % denaturing acrylamide was defined as 7 M urea and 40 %
formamide. Gradients of 38–48 % were used to separate
products amplified with universal primers. After migration of the PCR products at 85 V for 16 h, the gels were
stained with AgNO3 as described previously (Sanguinetti
et al., 1994).
PCR on the 16S rRNA gene of strain MucT was performed
with universal primers 11f and 1510r (Lane, 1991). The
following PCR programme was used: 94 uC for 5 min; 40
cycles consisting of 94 uC for 1 min 30 s, 48 uC for 30 s,
and 68 uC for 1 min 30 s; and finally 68 uC for 7 min. PCR
products were purified and concentrated with the Qiaquick
PCR purification kit (Qiagen) according to the manufacturer’s instructions. The purified 16S rRNA gene product
was sequenced on both strands using infrared Dye 41labelled primers 7f, 342r, 805f, 1100r and 1510r (Lane,
1991), and 968f (Nübel et al., 1996). One extra primer,
Muc1 (59-GGA AAC CCT GAT GGT GCG-39), which
targets a 339 bp specific region of the 16S rRNA gene
sequence of strain MucT, was designed to obtain unambiguous results. Sequences were automatically analysed on a
LI-COR DNA sequencer 4000L and corrected manually.
Pairwise sequence alignment was performed with the
program DNASTAR. The 16S rRNA gene sequence was
compared to sequences from GenBank using the program
BLASTN 2.0, available through the National Centre for
Biotechnology Information (NCBI) website (http://www.
ncbi.nlm.nih.gov/blast/). The ARB software package was
used to align cloned sequences and 16S rRNA gene
sequences of nearest relatives (Strunk & Ludwig, 1995). A
phylogenetic tree was constructed with ARB using the
neighbour-joining method. The distance matrix used in
the neighbour-joining method included stretches of sequence corresponding to Escherichia coli positions 63–1491.
Substrates and fermentation product concentrations were
determined in the culture before and after growth using
HPLC and GC methods as described previously (Stams
et al., 1993).
The use of serial dilution in an anaerobic medium containing mucin as energy source led to the isolation of a
predominant mucin-degrading bacterium from the human
http://ijs.sgmjournals.org
Fig. 1. DGGE profiles of the V6 to V8 regions of 16S rRNA
gene from strain MucT (lane 1), faecal sample (lane 2) and dilution 10”2 to 10”9 of the enrichment of faecal bacteria in mucin
medium (lanes 3–10). The arrowheads indicate the migration
position of the 16S rRNA gene amplicon of strain MucT.
faecal sample. As in all MPN studies, the low-dilution
cultures presumably gave rise to the fastest-growing organisms under the given culture conditions, whereas the highdilution cultures supported growth of the numerically
dominant organisms. In our study, a single band generated
by PCR-DGGE (Fig. 1) dominated all dilutions, indicating
that there was one predominant mucin-degrading bacterium and that this was also the fastest-growing mucindegrading organism. The MPN of mucin-degrading
organisms present in this faecal sample was estimated at
8?3±0?36109 (g faeces)21. No growth was observed in the
uninoculated mucin medium, indicating the sterility of
the mucin, nor in the medium inoculated with faecal
dilutions but not supplemented with mucin. This indicates
that growth occurred solely due to utilization of mucin by
the faecal bacteria.
Microscopic analysis of the enrichment dilutions revealed
that an oval-shaped organism was predominant in the first
dilutions of the enrichment. The 16S rRNA gene amplicons
from all the dilutions of the enrichment where growth was
observed (1022 to 1029) were analysed by DGGE (Fig. 1,
lanes 3–10). In the first dilutions, many bands were detected,
but one was predominant. This band became more intense
with increasing dilution and, in the highest dilution where
growth occurred (1029), this band was almost unique in
the profile. The faecal sample profile showed a band at the
same position, suggesting that the enriched micro-organism
1471
M. Derrien and others
containing this 16S rRNA gene is the same as the one
present in the faecal sample and represents at least 1 % of
the total intestinal bacterial community (Muyzer et al.,
1993). The mucin-degrading bacterium, whose 16S rRNA
gene corresponded to the major DGGE band present in
the enrichment (lane 10), was cultured from the highest
dilution with growth (1029) using the soft agar technique.
After 6 days, the most predominant colony type, white,
was grown in the mucin medium. White colonies were
picked, diluted in the mucin medium and transferred
into soft agar mucin medium. This purification step was
repeated twice. Finally, a single type of white colony
appeared. Phase-contrast microscopy revealed only one
morphotype, and the DGGE profile of the 16S rRNA gene
amplicon of the strain isolated showed the presence of a
unique band corresponding to the major band present in
the enrichment (Fig. 1, lane 1). The pure culture was
designated strain MucT.
An almost-complete 16S rRNA gene sequence of strain
MucT was determined (1433 bp). The most similar 16S
rRNA gene sequences, which were derived from studies of
uncultured colonic bacteria [HuCA18 and HuCC13 (Hold
et al., 2002) and L10-6 (Salzman et al., 2002)] were 99 %
identical to strain MucT. The cultured bacterium most
closely related to strain MucT was Verrucomicrobium
spinosum, and this was only distantly related (92 %). A
phylogenetic dendrogram based on 16S rRNA gene
sequences was constructed; it revealed that strain MucT is
related to the genera Prosthecobacter and Verrucomicrobium,
which are members of the order Verrucomicrobiales. Thus,
MucT belongs to the division Verrucomicrobia and the
class Verrucomicrobiae (Fig. 2). The majority of the
members of this new division are clones and only a few
are cultivated bacteria: a single genus, Verrucomicrobium
(Schlesner, 1987), after which the division was named;
four species of the genus Prosthecobacter (Prosthecobacter
debontii, Prosthecobacter dejongeii, Prosthecobacter fusiformis
and Prosthecobacter vanneervenii) (Staley et al., 1976;
Hedlund et al., 1997); Opitutus terrae (Chin et al., 2001)
and other ultramicrobacteria (Janssen et al., 1997); and the
recently described species Victivallis vadensis, the first
member of the division Verrucomicrobia to be isolated
from the GI tract (Zoetendal et al., 2003). Members of the
‘Verrucomicrobium’ group of bacteria have also been
identified in low numbers in human faeces-derived 16S
rRNA gene libraries (Wilson & Blitchington, 1996; Suau
et al., 1999; Hold et al., 2002). The division Verrucomicrobia
is composed of five subdivisions (Hugenholtz et al., 1998)
and the genera Prosthecobacter and Verrucomicrobium
are part of subdivision 1. Bacteria from these two genera
were isolated from freshwater habitats and are both
Gram-negative, aerobic and heavily fimbriated. Cells of
Verrucomicrobium have many prosthecae, whereas cells of
Prosthecobacter have only a single prosthecae. Strain MucT
shares some common characteristics; for example, it is
Gram-negative and can grow without vitamins. However,
strain MucT is distinct among the members of subdivision
1 in that it is strictly anaerobic and cells are oval-shaped in
contrast to the other members (see Table 1). On the basis
of a phylogenetic analysis, strain MucT does not belong
to the Verrucomicrobium or Prosthecobacter clusters and
Fig. 2. Phylogenetic tree showing the position of strain MucT among selected clones or strains belonging to the division
Verrucomicrobia. The tree, which was rooted using Escherichia coli as the outgroup, was generated by the neighbour-joining
method. The numbers before the interior branch points indicate the five major lineages within the division Verrucomicrobia as
proposed by Hugenholtz et al. (1998). Bar, 10 % sequence divergence.
1472
International Journal of Systematic and Evolutionary Microbiology 54
Akkermansia muciniphila gen. nov., sp. nov.
Table 1. Characteristics that differentiate the genus Akkermansia from other genera of the subdivision 1 of the division
Verrucomicrobia
Data taken from Schlesner (1987), Hedlund et al. (1997), Staley et al. (1976). +, Positive; 2, negative; W, weakly positive; ND, not determined. 1, strain MucT; 2, Verrucomicrobium (based on Verrucomicrobium spinosum); 3, Prosthecobacter (based on P. debontii, P. dejongeii,
P. fusiformis and P. vanneervenii). All genera are negative for motility, a requirement for vitamins, and growth on amino acids and other
organic acids. All genera are sensitive to ampicillin.
Characteristic
Cell morphology
Cell size (mm)
Tolerance to oxygen
Temperature range for growth (uC)
Prosthecate
Fimbriae
Capsule
Growth on:*
Glucose
Galactose
Fructose
Cellobiose
N-Acetylglucosamine
N-Acetylgalactosamine
Mucin
DNA G+C content (mol%)
1
2
3
Oval-shaped
0?6–1?0
2
20–40
2
2
+
Fusiform rod-shaped
0?8–1?061?0–3?8
+
26–34
+
+
Fusiform rod-shaped
0?562?0–8?0
+
1–40
+
+
ND
ND
WD
+
+
+
+
+
+
+
+/2d
+
+/2d
2
2
2
WD
WD
+
47?6
ND
ND
ND
ND
57?9–59?3
54?6–60?1
*Growth was determined by measuring OD600 in basal medium supplemented with the appropriate substrate (10mM final concentration;
see text).
DWhen a protein source is provided (peptone, yeast extract, tryptone and casitone at final concentration of 2 g l21 each).
dDepends on species.
should be considered as a separate phylogenetic branch. It
is therefore proposed that strain MucT represents a novel
species in a new genus belonging to subdivision 1 of the
Verrucomicrobia.
Strain MucT is an obligate chemo-organotroph. No growth
was detected on basal medium supplemented with vitamins
and purged with H2/CO2 (80 : 20). Rumen fluid and
vitamins were not required for growth on mucin and, for
further characterization of the strain, they were not added
to the mucin medium. Growth was not observed in mucin
medium in the absence of a reducing agent, as indicated by
the pink colour of the medium, demonstrating the strict
anaerobic nature of strain MucT. The isolate could grow
between 20 and 40 uC, with optimum growth at 37 uC. The
optimum pH for growth was 6?5. No growth was observed
below pH 5?5 or above pH 8. The doubling time of the
strain was approximately 1?5 h in mucin medium.
No growth was observed on glucose, cellobiose, lactose,
galactose, xylose, fucose, rhamnose, maltose, succinate,
acetate, fumarate, butyrate, lactate, casitone (0?5 %), Casamino acids (0?5 %), tryptone (0?5 %), peptone (0?5 %),
yeast extract (0?5 %), proline, glycine, aspartate, serine,
threonine, glutamate, alanine, N-acetylglucosamine or
N-acetylgalactosamine after 4 weeks incubation. Gastric
mucin isolated from human intestinal cell lines and adapted
http://ijs.sgmjournals.org
on 1025 M methotrexate (HT-29 MTX) to produce a
high amount of mucin (Lesuffleur et al., 1990) resulted in
growth of strain MucT to the same density as with hog
gastric mucin. Strain MucT could also grow on rich media,
Columbia and BHI, but with a final optical density of half
that of the mucin medium. No growth was observed on
rich WC anaerobe broth. When peptone, yeast extract,
tryptone and casitone (each at 2 g l21) were added to the
basal medium, growth was observed only when the sugars
N-acetylglucosamine, N-acetylgalactosamine and glucose
were added, although bacterial growth was less than a
quarter of that on mucin medium. When the solution
containing the nitrogen source was not added to the basal
medium supplemented with mucin, strain MucT could
grow to the same density, indicating that the isolate utilized
mucin as both carbon and nitrogen source. Strain MucT did
not produce H2, but acetate, propionate and ethanol were
formed from mucin fermentation. No sulfides were produced. Sulfates were released during fermentation of mucin
(0?71 mM), demonstrating sulfatase activity. It is presumed
that the limited ability of strain MucT to grow on the many
substrates tested may be due to the complex structure of
the mucin, which is composed of both oligosaccharides
and amino acids, and that strain MucT requires a combination of all these components to reach a high density.
It is likely that strain MucT produces one or more
1473
M. Derrien and others
(a)
(b)
Fig. 3. EM images of strain MucT. (a) SEM image. Bar, 1 mm. (b) TEM image of a negatively stained preparation. Note the
thickened but extensive capsule fibres of the cells. Bar, 0?5 mm.
appropriate glycosidases to degrade the N-acetylgalactosamine
and N-acetylglucosamine components from mucin, which
might be exposed in the terminal part, and to use them as
growth substrates.
Cells of strain MucT were oval-shaped (Fig. 3a), showing a
different size depending on the medium. In mucin medium,
strain MucT was 640 nm in diameter and 690 nm in length
and in BHI, strain MucT was 830 nm in diameter and 1 mm
in length. Cells stained Gram-negative. Flagella were not
seen on negatively stained EM preparations. Spore formation was never observed. In mucin medium, the organism
could grow as single cells or in pairs, but rarely in chains;
it often formed aggregates in which a translucent layer of
material was observed between organisms. In BHI and
Columbia media, this material was rarely, if ever, observed
and cells occurred singly or in pairs, but rarely in groups.
In basal medium, supplemented with N-acetylglucosamine
or N-acetylgalactosamine, together with some sources of
proteins (a combination of yeast extract, peptone, tryptone
and casitone), cells occurred singly and sometimes in pairs.
Cells of strain MucT grown in mucin medium could exclude
Indian ink, which is characteristic of capsule-possessing
bacteria. EM revealed the existence of filamentous structures
on cells grown in mucin medium (Fig. 3b). It is assumed
that these filaments are capsular polymers that are used
to connect cells together. Since this aggregation is mainly
observed in mucin medium, this capsule may aid in adhesion and colonization of mucin-secreting epithelia in the
GI tract. On soft agar medium, colonies of strain MucT
appeared white and were 0?7 mm in diameter.
Based on morphological, physiological and phylogenetic
features of strain MucT, a new genus, Akkermansia, with
the type species Akkermansia muciniphila gen. nov., sp. nov.
is proposed.
Description of Akkermansia gen. nov.
Akkermansia (Ak.ker.man9si.a. N.L. fem. n. Akkermansia
derived from Antoon Akkermans, a Dutch microbiologist
recognized for his contribution to microbial ecology).
1474
Cells are oval-shaped, non-motile and stain Gram-negative.
Strictly anaerobic. Chemo-organotrophic. Mucolytic in
pure culture.
The type species is Akkermansia muciniphila.
Description of Akkermansia muciniphila sp. nov.
Akkermansia muciniphila (mu.ci.ni9phi.la. N.L. neut. n.
mucinum mucin; Gr. adj. philos loving; N.L. fem. adj.
muciniphila mucin-loving).
Cells are oval-shaped, non-motile and stain Gram-negative.
The long axis of single cells is 0?6–1?0 mm, depending on
the substrate used. Cells occur singly, in pairs, in short
chains and in aggregates. Growth occurs at 20–40 uC
and pH 5?5–8?0, with optimum growth at 37 uC and
pH 6?5. Strictly anaerobic. Able to grow on gastric mucin,
brain–heart infusion and Columbia media, and on Nacetylglucosamine, N-acetylgalactosamine and glucose
when these three sugars are in the presence of (each at
2 g l21) peptone, yeast extract, casitone and tryptone.
Cellobiose, lactose, galactose, xylose, fucose, rhamnose,
maltose, succinate, acetate, fumarate, butyrate, lactate,
casitone, Casamino acids, tryptone, peptone, yeast extract,
proline, glycine, aspartate, serine, threonine and glutamate
do not support growth. Capable of using mucin as carbon,
energy and nitrogen source. Able to release sulfate in a free
form from mucin fermentation. In mucin medium, cells are
covered with filaments. Growth occurs without vitamins.
Colonies appear white with a diameter of 0?7 mm in soft
agar mucin medium.
The type strain is MucT (=ATCC BAA-835T=CIP
107961T), isolated from the human intestinal tract. Its
DNA G+C content is 47?6 mol%.
Acknowledgements
The authors are grateful to E. Tosi-Couture (Institut Pasteur, Unité
Toxines et Pathogénie bactériennes, Paris, France) for performing
the TEM. Dr G. Huet (INSERM, U560, Lille, France) is gratefully
acknowledged for the gift of the HT-29 MTX mucin. We thank E. G.
International Journal of Systematic and Evolutionary Microbiology 54
Akkermansia muciniphila gen. nov., sp. nov.
Zoetendal for discussion and for critically reading the manuscript and
H. Smidt for help in the phylogenetic analysis. We thank Professor
Dr H. G. Trüper for his help regarding the Latin nomenclature. This
work has been carried out with the financial support of the European
Community specific RTD programme ‘Quality of Life and Management of Living Resources’ research project EU & Microfunction
(QKL1-2001-00135). C. M. P. was supported by the Research Council
for Earth and Life Sciences (ALW) with financial aid from the
Netherlands Organization for Scientific Research (NWO).
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