REVIEWS

Magnetosome biogenesis in
magnetotactic bacteria
René Uebe and Dirk Schüler
Abstract | Magnetotactic bacteria derive their magnetic orientation from magnetosomes, which
are unique organelles that contain nanometre-sized crystals of magnetic iron minerals. Although
these organelles have evident potential for exciting biotechnological applications, a lack of
genetically tractable magnetotactic bacteria had hampered the development of such tools;
however, in the past decade, genetic studies using two model Magnetospirillum species have
revealed much about the mechanisms of magnetosome biogenesis. In this Review, we highlight
these new insights and place the molecular mechanisms of magnetosome biogenesis in the
context of the complex cell biology of Magnetospirillum spp. Furthermore, we discuss the diverse
properties of magnetosome biogenesis in other species of magnetotactic bacteria and consider
the value of genetically ‘magnetizing’ non-magnetotactic bacteria. Finally, we discuss future
prospects for this highly interdisciplinary and rapidly advancing field.

Microoxic
Magnetotactic bacteria are ubiquitously present in
A concentration of oxygen that aquatic sediments and can be enriched from these sed-
is less than 21% and therefore iments by taking advantage of their directed motility
lower than that found in the in the presence of magnetic fields. The basis for this
atmosphere.
intriguing behaviour is the presence of magnetosomes,
Biomineralization which are unique organelles that are composed of
The process by which membrane-enveloped crystals of a magnetic iron min-
organisms form inorganic eral, and the alignment of these organelles along the
minerals. cellular motility axis by dedicated cytoskeletal struc-
tures. Species of magnetotactic bacteria are found in
diverse Gram-negative phylogenetic groups, includ-
ing the Alphaproteobacteria, Gammaproteobacteria,
Deltaproteobacteria and Nitrospira classes and the candi-
date phyla Latescibacteria (also known as candidate divi-
sion WS3) and Omnitrophica (also known as candidate
division OP3) of the Planctomycetes–Verrucomicrobia–
Chlamydiae (PVC) bacterial superphylum1–7. All species
of magnetotactic bacteria are highly motile and share a
microoxic or anoxic lifestyle that is specifically adapted to
the oxygen and redox gradients in the stratified freshwater
or marine sediments that they commonly inhabit. It is
now established that magnetosomes function as naviga-
tional devices for orientation in the Earth’s magnetic field
through close integration with other environmental cues
University of Bayreuth,
(BOX 1), although they have also been suggested to function
Department of Microbiology,
Universitätsstraße 30, as detoxifiers of metal ions or reactive oxygen species, as
95447 Bayreuth, Germany. gravity sensors, or as ‘geobatteries’ that gain energy from
Correspondence to D.S.  redox cycling 8,9. Despite a common function in mag-
dirk.schueler@uni-bayreuth.de netic orientation, the mineral composition (magnetite or
doi:10.1038/nrmicro.2016.99 greigite), shape and intracellular arrangement of magne-
Published online 13 Sep 2016 tosomes can vary substantially between different species.
The study of magnetosome biogenesis is a growing
interdisciplinary area of research that encompasses fields
as diverse as microbiology, geobiology, biophysics, bio-
technology and nanotechnology. Since the first report of
magnetotactic bacteria10 and their serendipitous redis-
covery more than four decades ago11, many studies have
examined the physiology, ecology and phylogeny of these
organisms, as well as the magnetic and mineral properties
of magnetosomes, and these studies have been described
in numerous excellent reviews8,12–16. However, because
of a historical lack of genetically tractable laboratory
models, fewer advances had been made in elucidating
the genetic determinants and molecular mechanisms
of magnetosome biogenesis. Fortunately, during the
past 10 years or so, tremendous progress has been made
in the molecular analysis of magnetosome biogenesis,
and the study of magnetotactic bacteria has consequently
emerged as a powerful model for the more general study
of the magnetic orientation of organisms, biomineralization
processes and prokaryotic organelle formation.
Much of this progress has relied on the development
of methods for the cultivation, at various scales, and
the genetic manipulation of Magnetospirillum magne­
ticum AMB‑1 (AMB) and Magnetospirillum gryphis­
waldense MSR‑1 (MSR), which has enabled them to be
used as model strains of magnetotactic bacteria in the
laboratory. These strains, both of which are from
the Alphaproteobacteria class and produce single chains
of cuboctahedral magnetite (Fe3O4) crystals, have the
advantage of a relative ease of cultivation in the labo-
ratory. Studies that have used AMB and MSR as model

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Box 1 | Magneto-aerotaxis
What is the function of magnetosome chains? As first suggested at the
time of their discovery in 1975 (REF. 11), magnetosome chains are cellular
sensors for magnetotaxis, which is widely assumed to occur when
magnetotactic cells that are passively aligned along magnetic field
lines actively swim11, although a recent study suggests that some
magnetotactic bacteria might sense not only the direction but also the
gradients of magnetic fields138. The benefit of magnetotaxis is probably
that alignment to geomagnetic fields reduces three-dimensional
swimming, which is the form of swimming that is used by most motile
bacteria, to a linear movement along vertical inclines that are chemically
stratified. Indeed, magnetotaxis seems to be deeply integrated
with chemotactic responses, notably aerotaxis, and integration with
phototaxis has also been observed139,140. In ‘magneto-aerotaxis’,
magnetotactic bacteria in stratified environments are efficiently guided
to their preferred oxygen concentration, which is close to, or below, the
oxic–anoxic transition zone66,141.
Polarity
Under natural conditions, the magneto-aerotaxis of most isolates of
magnetotactic bacteria is ‘polar’; that is, the predominant swimming
direction of these bacteria in the presence of oxygen
is north‑seeking in the Northern Hemisphere but Northern Hemisphere
south‑seeking in the Southern Hemisphere13. However,
magnetotaxis in laboratory strains of Magnetospirillum spp.
lacks north–south polarity: although motility in these
Water column
conditions, which are the main environmental cues for swimming in this
bacterium, such that oxygen upshifts are able to cause a collective and
instantaneous reversal of the north‑seeking swimming polarity of
anoxic-adapted cells142. However, as bacterial cells are usually
morphologically symmetrical (for example, with cell shapes that are
spirals or rods), how magnetotactic bacteria adopt a cellular polarity is
not known.
Integration with chemotaxis
The genomes of most magnetotactic bacteria encode exceptionally high
numbers of proteins that are predicted to have functions in chemotaxis or
signal transduction, which might reflect a variable and fine-tuned range
of responses that have not yet been explored. For example, MSR has at
least 56 genes that encode putative chemoreceptors, compared with only
five such genes in Escherichia coli, and four copies of the canonical
chemotaxis operon, compared with a single copy in E. coli145. Although
the functions of three of these operons are unknown, deletion of the
fourth operon abolished both aerotaxis and the polarity of magnetotaxis
in MSR, thereby providing the first evidence that magnetotaxis can be
integrated with chemotaxis through a common sensory pathway142.
Southern Hemisphere
Magnetic O2
N S
torque

bacteria is aligned to magnetic field lines, swimming is not CCW CCW

preferentially directed towards either pole . The lack of
139 + + Oxic
polarity probably results from the loss of selection during
NS SS
laboratory cultivation by shaking, which eliminates oxygen – –
gradients, as laboratory evolution experiments with CW CW
Magnetospirillum gryphiswaldense MSR‑1 (MSR) show that
predominant north-seeking or south‑seeking swimming
polarity is restored when strains are cultured without + + + + + + Micro
shaking and in the presence of vertical oxygen gradients oxic
Sediment

and parallel or antiparallel magnetic fields 142,143

(these
strains can then be used to study the genetics of polarity; – – – – – – Oxic–
anoxic
see below). Similarly, an ovoid species of magnetotactic transition
bacteria, the marine bacterium MO‑1, can lose, acquire or zone
reverse the polarity of magneto-aerotaxis, depending on
Anoxic
the direction of the redox gradient and magnetic field144, S N
and in only a few generations, which may argue for an
epigenetic mechanism of inheritance142. Finally, the polarity Geomagnetic field line Inclination
of magnetotaxis in MSR populations that have reacquired
CCW, counterclockwise; CW, clockwise; N, north; S, south
polarity can be reversed by changes in oxygen and redox

Nature Reviews | Microbiology

Aerotaxis
Movement of an organism
towards (positive) or away
(negative) from oxygen.
Chemoreceptors
Proteins that detect certain
chemical stimuli in the
environment.
magnetotactic bacteria have shown that magneto-
some biogenesis comprises four steps (FIG. 1a–f). First,
the magnetosome membrane is invaginated from the
cytoplasmic membrane, forming either vesicle-like
permanent invaginations or detached vesicles. Second,
either before, coincidently or following invagination,
magneto­s ome proteins are sorted to the magneto-
some membrane. Third, iron is transported into the
magneto­some membrane invagination or vesicle and
mineralized as magnetite crystals. Fourth, a magneto-
some chain is assembled and positioned for segregation
during cell division.
In this Review, we describe the factors that mediate
each of the four stages of magnetosome formation in
magnetotactic bacteria. Although our primary focus
is magnetosome biogenesis in AMB and MSR, we also
highlight the diversity of magnetosomes that have been
observed in other species of magnetotactic bacteria.
Furthermore, we discuss the use of genetic engineering
to transfer the capacity for magnetosome biogenesis to
other organisms to generate synthetic magnetic micro-
organisms, and we consider the potential biotechno-
logical applications of magnetic microorganisms and
of magnetosomes. Finally, we address open questions of
magnetosome biogenesis and discuss new perspectives
and directions for future research in the field.
Genetics of magnetosome biogenesis
All four steps of magnetosome biogenesis are gov-
erned by complex machinery that has been shown
to comprise a large number of genetic determinants.
As yet, more than 40 different genes have been iden-
tified that encode magnetosome-associated proteins
and/or that yield ­magnetosome-related phenotypes in
Magnetospirillum spp. when mutated (Supplementary
information S1 (table)). Although several auxiliary,

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a d

b e

c f

g
mms6 operon mamGFDC mamAB operon mamXY operon feoAB1
operon operon

mms48 mms36 mamD mamH mamI mamE mamJ mamK mamM mamN mamO mamP mamQ mamB mamU ftsZ-like mamZ mamX mamY feoA1 feoB1
mmsF mamF mamC mamL mamA mamR mamS
mms6 mamG mamT
1 kb
Hypothetical protein ArsB/NhaD family permease PDZ domain-containing protein
MTB/Magnetospirillum specific Actin-like protein Tetratricopeptide repeat domain
Sphingosine kinase related Tubulin-like protein Serine family protease
Major facilitator family transporter LemA family protein FeoA family protein
Cation diffusion facilitator family transporter Magnetochrome domain-containing protein FeoB family protein

Figure 1 | Overview of the mechanisms and genetics of magnetosome biogenesis. A model for magnetosome
Nature Reviews | Microbiology
biomineralization and chain formation in Magnetospirillum gryphiswaldense MSR‑1 (MSR), which is one of the strains of
magnetotactic bacteria that has been most intensively studied in the laboratory (parts a–f). According to this model,
magnetosome biogenesis is induced in an ‘empty’ cell that is entirely devoid of magnetosome membranes and magnetite
crystals (part a). Magnetosome vesicles (red circles) are then formed throughout the cell by invagination of the
cytoplasmic membrane, which become pinched off at later stages of biogenesis (part b). Iron is transported into
magnetosome membrane vesicles and magnetite nucleation leads to biomineralization of cuboctahedral magnetite
crystals (dark grey octahedrons), with a single crystal present in each mature magnetosome (part c). Magnetosomes are
aligned into chains through the interaction of MamJ (yellow star) on the magnetosome membrane with a filament
(green dashed line) that is formed through the polymerization of MamK, an actin-like protein (parts c and d). MamK is also
required for the positioning of the magnetosome chain at mid-cell. Organization of magnetosomes into chains helps to
align individual magnetic dipoles (white arrows) to generate a strong dipole that can ‘sense’ the geomagnetic field (part
d). Cell division is initiated through asymmetric constriction of the FtsZ ring (light grey circle; part e); during cell division,
unidirectional indentation of the cell wall in conjunction with cell kinking helps to bend the magnetosome chain and
thereby reduce magnetostatic forces to promote an even segregation of the magnetosome to the daughter cells (parts e
and f). After the completion of cell division, magnetosome chains are rapidly repositioned (black arrows) from the new cell
pole along the MamK filament to the mid-cell (part f). Molecular organization of the genomic magnetosome island (MAI)
of MSR, which contains genes that have functions specific to magnetosome biogenesis that are organized into five
operons. Genes are colour-coded according to characteristic features of the encoded proteins (part g). FeoA, ferrous iron
transport protein A; MTB, magnetotactic bacteria.

non-essential functions are contributed by general met-
abolic and regulatory cellular pathways that are not spe-
cific to magnetosome biogenesis, all specific functions
in magnetosome biogenesis are associated with a set
of about 30 mam (magnetosome membrane) and mms
(magnetic particle-membrane specific) genes17–20 that are
clustered in a single chromosomal region, the genomic
magneto­some island (MAI)21,22. The MAI extends across
about 100 kb, which is equivalent to approximately 2%
of a typical Magnetospirillum spp. genome and shares

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Genomic islands
Discrete genomic regions that
can be transferred horizontally.
These genomic regions
typically encode several
transposases and integrases
to increase the rate of DNA
turnover and are frequently
associated with the adaptation
of microorganisms to new
environments.
a (including the LemA domain and the tetra­tricopeptide
OM
CM
AMB
OM
CM
characteristics with other bacterial genomic islands (such
as putative insertion sites near tRNA genes, distinct
levels of GC enrichment, and a large number of mobile
genetic elements)21,23,24. MAI genes are organized into
five poly­cistronic operons (FIG. 1 g): the large (16–17 kb)
mamAB operon and the small feoAB1, mamGFDC,
mms6 and mamXY operons. The mamAB operon con-
tains several genes that are involved in key steps of mag-
netosome biogenesis (FIG. 1 g), and this operon is essential
and sufficient to sustain at least some rudimentary bio­
mineralization. By contrast, the four small operons
encode non-essential proteins that have accessory roles
in regulating the biomineralization of crystals of defined
size and shape (including the putative iron transporters
FeoAB1 and MamZ; the tubulin-like protein FtsZm;
and the magneto­tactic bacteria-specific proteins MamC
(also known as Mms13), MamD (also known as Mms7),
MamF, MamG, MamX, MamY, Mms6, MmsF, Mms36
and Mms48)25–28. Several genes in the mamAB operon
encode proteins that belong to well-­characterized fam-
ilies (including the actin-like superfamily, the c­ ation
diffusion facilitator (CDF) family and the HtrA-like pro-
tease family) or that contain functional domains that are
also found outside of species of magnetotactic bacteria
repeat (TPR) domain; FIG. 1 g). In addition to genes
that are essential for magnetosome biogenesis, the
mamAB operon contains genes that produce no pheno­
type, or only a weak phenotype, when deleted; these
non-­essential genes include mamU and mamV. AMB,
but not MSR, has an additional MAI operon, limJOE
(­mamJOE-like), which contains non-essential para­
logues of mamJ, mamO and mamE24,29,30, and an addi-
tional ‘magnetosome islet’, which is a smaller genomic
region that contains non-essential paralogues of mamE,
mamJ, mamK, mamL, mamM, mamF and mamD31.

Formation of the magnetosome membrane

MSR Compartmentalization of magnetosome biogenesis
b by the magnetosome membrane provides the grow-
OM ing magnetite crystal with a specialized ‘nano-­reactor’
Periplasm in which the iron, redox and pH environments of
M C D H
B L Q I E O Z G X F
CM bio‑mineralization can be strictly regulated. The mag-
netosome membrane was shown to comprise a 5–6 nm
GFDC

E E E proteinaceous phospholipid bilayer that is invaginated

ZHX
MamJ
MamA
FtsZm
Figure 2 | Magnetosome membrane invagination and protein recruitment.
Nature Reviews | Microbiology
a | Cryoelectron tomogram sections that show representative vesicle-like invaginations
of the magnetosome membrane in Magnetospirillum magneticum AMB‑1 (AMB) and
magnetosome membrane vesicles in Magnetospirillum gryphiswaldense MSR‑1 (MSR),
which are a mixture of empty vesicles and vesicles that contain electron-dense magnetite
crystals. Invaginations of the cytoplasmic membrane are only rarely seen in MSR
(white arrow), which suggests that vesicles are rapidly separated from the cytoplasmic
membrane in these cells, but the magnetosome membrane in AMB is observed as
vesicle-like invaginations that are still connected to the cytoplasmic membrane at all
stages of biomineralization. b | Model of magnetosome membrane invagination and
protein recruitment. The presence and interaction of several magnetosome proteins are
required to induce membrane curvature and vesicle formation. It has been suggested that
MamB might be a landmark protein for the formation of protein interaction networks
that induce membrane curvature. Assembly of protein complexes on the magnetosome
membrane, and the targeting of additional proteins to the magnetosome membrane,
is facilitated by MamA, MamE and possibly FtsZm (single letters denote respective
Mam proteins). CM, cytoplasmic membrane; OM, outer membrane. Images in the
top panels of part a are adapted with permission from REF. 34, AAAS. Images in
the bottom panels of part a are adapted from REF. 37.
from the cytoplasmic membrane32–35 (FIG. 2a). In both
MSR and AMB, these invaginations originate simulta-
neously from several nonspecific cellular locations, as
was revealed by tracking de novo magnetosome bio-
genesis by time-lapse fluorescence microscopy and
cryoelectron tomography, using inducible gene expres-
sion systems to ensure synchronicity36,37. In MSR, the
invagination of magnetosome membranes is likely to
proceed rapidly as no initial stages of membrane defor-
mation have been detected37. The formation of the
magnetosome membrane is independent of magnetite
biomineralization, as shown by the presence of empty
vesicles in iron-starved cells or biomineralization-
defective mutants34–36,38,39 (Supplementary information
S1 (table)), although, once formed, the growth of the
magnetosome membrane in AMB has been shown to
occur in a biomineralization-dependent manner, which
suggests the presence of a biomineralization-dependent
checkpoint 36. By contrast, mutants that are unable to
form magnetosome vesicles are also unable to form mag-
netite, which indicates that the formation of magneto-
some vesicles is a requirement for biomineralization. In
AMB cells, nearly all magnetosome vesicles seem to be
connected to the cytoplasmic membrane as permanent
invaginations34, whereas invaginating magnetosomes
were only rarely detected in MSR cells35, which sug-
gests that in this strain the invaginating magnetosome

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Cation diffusion facilitator
(CDF). A ubiquitous family of
proteins that transport divalent
metal cations (such as Zn2+,
Mn2+, Cd2+, Co2+ and Fe2+)
using a proton or potassium
antiport mechanism.
Tetratricopeptide repeat
(TPR). A structural motif of 34
degenerate amino acids that is
often involved in the assembly
of multiprotein complexes by
mediating protein–protein
interactions.
Landmark protein
Proteins that mark certain
cellular regions (or structures)
and that may direct the
assembly of larger multiprotein
complexes.
membrane becomes pinched off into magnetosome vesi-
cles at later stages of biogenesis37,40 (FIG. 2a; for simplicity,
we use ‘vesicles’ to refer to both vesicles and v­ esicle-like
invaginations). Using electron microscopy, the magne-
tosome membrane has been visualized as vesicles with
diameters of approximately 10–70 nm in both MSR
and AMB33–38. Finally, the magnetosome membrane
has a very similar lipid composition to the cytoplasmic
membrane, but has a distinct set of proteins, which are
encoded by mam and mms genes17,18.
Invagination of the magnetosome membrane. The
ability to bend and remodel a lipid bilayer is generally
thought to depend on specific proteins that provide the
energy required to generate and sustain membrane cur-
vature through membrane scaffolding 41 or through the
insertion of amphipathic helices into the membrane42.
However, no MAI gene has obvious homology to known
determinants of membrane remodelling in eukaryotes.
Previously, mamB, mamL, mamI, and mamQ had been
implicated in magnetosome membrane invagination24,
as AMB deletion mutants of these genes seemed to lack
magnetosome membrane vesicles. However, careful
analysis of MSR deletion mutants that lacked mamI or
mamL recently revealed the presence of vesicles that
were indistinguishable from those formed in wild-
type cells. Furthermore, when grown at low tempera-
tures that favour biomineralization, MSR mutants that
lacked mamI or mamL were capable of forming vesicles
that contained small haematite (a non-magnetic fer-
ric oxide of Fe3+) or magnetite crystals, respectively. In
addition, aberrant electron-dense vesicular structures
that seemed to be devoid of mineral particles could be
readily observed in MSR deletion mutants that lacked
mamQ or mamL, and even in a MamM-deletion mutant
that also contains magnetosome membrane vesicles
indistinguishable from those found in wild-type cells,
as shown by previous experiments37. Thus, in mutants
that lack mamL or mamM, both small, empty vesicles
akin to those formed in wild-type cells and dense mag-
netosome membrane-like structures are present 37,43.
Together, these data from genetic studies indicate that
MamI, MamL, MamM and MamQ are not essential for
magnetosome membrane invagination in MSR, but that
instead these proteins have functions in the biominer-
alization of magnetite and the maturation of magne-
tosome membrane vesicles37. Similarly, elimination of
MamY in MSR or AMB had only weak effects on mag-
netosome membrane biogenesis in vivo27,44, even though
this protein has been reported to bind to and tubulate
membranes in vitro44.
Although MamM, MamL, MamI, MamQ, MamY
and other factors may have non-essential roles in the
invagination of the magnetosome membrane, genetic
findings suggest that MamB is the most crucial protein
for this process24,37,43. Indeed, although MamB has a
suspected function in magnetosomal iron uptake (see
below), several findings also indicate a role for MamB
in magnetosome membrane invagination. First, dele-
tion of mamB caused more severe aberrant magneto-
some membrane phenotypes than deletion of other
magnetosome-associated genes, as vesicle-like struc-
tures were only rarely observed in these mutants in
MSR, and entirely absent in AMB24,37,43. Second, MamB
interacts with proteins that are found on the magneto-
some membrane that are involved in later steps of mag-
netosome biogenesis, which suggests that MamB might
be a landmark protein that captures other proteins to
induce invagination24,43. However, genetic findings also
indicate that mamB alone is insufficient to invaginate
the magnetosome membrane. Expression of mamBLIQ
alone failed to induce invagination of the magnetosome
membrane in an AMB MAI-deletion mutant 24, and
expression of a synthetic operon comprising a subset
of seven key magnetosome genes (mamLQBIEMO)
failed to induce invagination of the magnetosome
membrane in the MSR‑1B strain, which lacks the
mamAB, mamGFDC and mms6 operons37 (although
mamLQBIEMO was sufficient to restore the formation
of intracellular membranes in MSR cells in which only
the large mamAB operon had been deleted)37. These
findings suggest that although invagination of the
magnetosome membrane depends on the presence of
MamB, possibly through suggested roles in recruiting
other proteins, these proteins alone are insufficient to
invaginate the magnetosome membrane37. Therefore, a
critical number of proteins from mamLQBIEMO and
a larger set of magnetosome-associated proteins seem to
be required in addition to MamB (FIG. 2b), even though,
probably owing to functional overlap, loss of function of
each of these proteins individually has no effect on the
invagination of magnetosome membranes. The contri-
bution of these additional proteins to the invagination
of magnetosome membranes might involve the forma-
tion of larger protein complexes that generate a lateral
pressure to drive membrane bending, similarly to the
membrane bending that is induced by protein crowd-
ing 45 (FIG. 2b). According to this model, the biogenesis of
magnetosome membrane vesicles occurs continuously at
several nonspecific cellular locations, which suggests that
the invagination of magnetosome membranes is a far less
regulated process than the invagination of membranes
during vesicle formation in eukaryotic cells46.
Targeting proteins to the magnetosome membrane.
All Mam and Mms proteins that have been studied in
detail have been found to associate with purified mag-
netosomes17,18,27,47,48, although the quantity that is present
in purified magnetosome preparations varies for each
protein. Furthermore, localization to the magnetosome
membrane has been confirmed for many of these pro-
teins by immunolabelling or fluorescence tagging in
intact cells24,29,39,43,49,50. The abundance of MamA, MamP,
MamY and several other proteins on the magnetosome
membrane seems to fluctuate, possibly even between
individual magnetosomes that are at various stages of
maturation but located in a single chain39,44,51. However,
how proteins are specifically targeted to the magne-
tosome membrane is not currently known, and no
motifs that encode sorting signals to the magnetosome
membrane have been identified. In addition to trans­
membrane proteins, such as MamC, MamE and MamM,

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PDZ domain
A short (80–90 amino acids)
protein domain that is often
involved in organizing signalling
complexes by forming protein–
protein interactions with
carboxy‑terminal peptides. The
name derives from the proteins
in which the domain was first
described: postsynaptic
density protein 95 (PSD95),
discs large homologue 1(Dlg1)
and zonula occludens 1 (ZO‑1).
Mössbauer spectroscopy
A spectroscopic technique that
is based on the Mössbauer
effect, which is the nearly
recoil-free, resonant absorption
and emission of gamma rays in
solids.
the magnetosome membrane is decorated with several
soluble proteins that are thought to bind to the mem-
brane indirectly using the TPR motif of MamA or the PDZ
domain of MamE or MamP, which are known to medi-
ate protein–protein interactions. Accordingly, MamA,
MamC, MamF and MamI became mis­localized in AMB
cells that lacked MamE29. MamA forms a large globular
scaffold that surrounds magnetosomes and that assem-
bles by a unique mechanism based on self-­recognition
of TPR motifs52,53 (FIG. 2b), which has been proposed
to facilitate further protein–protein interactions and
‘activate’ magnetosomes for magnetite biomineraliza-
tion39,52. Consistent with a role for MamA in recruiting
other proteins to the magnetosome membrane, MamA
was recently found to interact with Mms6 (REF. 54), and
MamC, which is one of the most abundant proteins on
magnetosome membranes in wild-type cells, was mis­
localized in MSR mamA-deletion mutants55. The stabil-
ity of several other magnetosome-associated proteins is
also dependent on interactions with other proteins
on the magnetosome membrane. For example, the
formation of a heterodimer with MamM is required
for the stabilization of MamB in MSR43 (FIG. 2b). During
the assembly of protein complexes on the magneto-
some membrane, the magnetotactic bacteria-specific
magnetite-binding proteins Mms5, Mms6 and MamD
undergo proteolytic processing, so that only their acidic
carboxy‑­terminal ends remain associated with magneto­
somes20,56–59. Remarkably, for Mms6 and some other
small, highly abundant proteins, such as MamC, MamF,
MmsF and MmxF, spontaneous self-assembly and oli-
gomerization have been reported in in vitro assays that
lack detergents and lipids56,60,61, although the relevance
of this multi­merization for the assembly of protein com-
plexes on magnetosome membranes in vivo remains to
be shown. Together, studies of the protein composition
of magnetosome membranes suggest that protein tar-
geting to the magnetosome membrane is a hierarchical
process that involves several different mechanisms that
remain poorly understood.
Biomineralization of magnetite crystals
Iron uptake. Following the formation of the magneto-
some membrane, a process of biomineralization occurs
in which large amounts of iron are accumulated in mag-
netosome membrane vesicles. Depending on the growth
conditions, more than 99.5% of the intracellular iron
in MSR may be present in magnetosomes, which can
account for more than 4% of the dry weight of the cell12,18.
Intriguingly, magnetite biomineralization is already
detectable at extracellular iron concentrations that are
growth limiting (less than 1 μM), even though in vitro
studies suggest that this biomineralization requires a
local supersaturated concentration of iron as high as
30 mM12,62. Furthermore, iron uptake and magnetite
biomineralization were found to become saturated63,64
at extracellular iron concentrations that remained far
less than 30 mM (approximately 20–50 μM) but that
were equivalent to the environmental abundance of sol­
uble iron in the sediment layers in which magnetotactic
­bacter­ia are most numerous65.
Three possible routes have been proposed for iron
uptake into magnetosome membrane vesicles (FIG. 3a).
In the first proposed route, iron enters the vesicle lumen
when the magnetosome membrane is still in physical
contact with the cytoplasmic membrane, by direct trans-
port or diffusion from the periplasm66. An alternative,
but not mutually exclusive, route entails the uptake
of Fe2+or Fe3+ into the cell using general (but species-
specific) cellular iron transport systems and subsequent
iron transport across the magnetosome membrane by
magnetosome-specific transporters28,43,67. Finally, based
on data from Mössbauer spectroscopy, iron for mag-
netite biomineralization has also been proposed to be
directly transported from the cytoplasmic membrane to
the magnetosome membrane by ligation to unknown
organic substrates, which would be released from the
cytoplasmic membrane at the interface with the mag-
netosome membrane; such a mechanism would enable
biomineralization to occur without the transport of
free iron through the cytoplasm40. Thus, although the
same iron transporters might be used for initial trans-
port across the cytoplasmic membrane, the intracellular
pathways for magnetite formation are proposed to be
distinct from other cellular iron transport pathways40.
In agreement with an iron uptake mechanism that is
specific to magnetosomes, general iron uptake and
homeostasis systems are only poorly conserved across
different species of magnetotactic bacteria. For example,
the production of siderophores, which has been reported
in AMB and a few other species of magnetotactic bac-
teria68–70 but not MSR63,71, is probably not relevant for
magnetite biomineralization. Further support for the
existence of such a mechanism comes from genetic evi-
dence that indicates that general iron uptake systems
are distinct from magnetite biomineralization. This
evidence involves the deletion of ferric-uptake regula-
tor (fur) or irrB, which encode members of the iron-
responsive Fe3+ uptake regulator family that regulate the
transcription of all known general iron uptake systems
in MSR71,72. Deletion of fur or irrB caused only weakly
defective magnetite biomineralization phenotypes71–73
and the expression of magnetosome-associated genes
was not affected in these mutants, which indicates that
magnetite biomineralization does not substantially rely
on general cellular iron uptake systems.
Currently, most evidence supports the notion that
iron accumulation in magnetosome membrane ves-
icles occurs subsequently to iron uptake by the cell.
For example, in MSR, loss of the GTP-dependent Fe2+
transporters FeoB1 and FeoB2, which are probably dis-
tributed over the entire cytoplasmic membrane and not
specific to the magnetosome membrane, caused a strong
impairment of biomineralization. As these receptors are
thought to transport ferrous iron from the periplasm
to the cytoplasm, this finding suggests that ferrous iron
for magnetite formation is initially transported into the
cytoplasm1,25,74. Subsequently, iron is transported from
the cytoplasm into magnetosome membrane vesicles in
a step that is assumed to be mediated by MamB and
MamM, which are members of the Fe/Zn‑transporting
subfamily of divalent metal CDF proteins43,75,76, and

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a Fe2+ Fe3+ OM ◀ Figure 3 | Biomineralization of magnetite crystals.

Organic substrate
a | Models of iron uptake in magnetotactic bacteria. One
Periplasm
? suggested model of iron uptake by magnetosome vesicles
CM proposes that extracellular iron is imported into the
Fe2+ transporter Fe2+
Fe2+ Fe3+ transporter cytoplasm by cytoplasmic membrane transporters and is
subsequently transported into magnetosome membrane

M
Fe2+ Fe2+ Fe3+

ZH
vesicles by magnetosome-specific transporters (probably

B
by MamB and MamM for Fe2+, and MamH and MamZ for
Fe3+; single letters denote respective Mam proteins). In a
Biochemical Fur Biochemical second model, iron is ligated to organic substrates directly
Fe2+ metabolism FeOx FeR Fe3+ metabolism after transport across the cytoplasmic membrane and
released into the magnetosome without the transport of
free iron through the cytoplasm, whereas a third model
proposes that iron import into vesicles might occur directly
b from the periplasm of the cell envelope. The intracellular
O2 NO3– Maturation
N2 Nucleation Fe2+/Fe3+ ratio is regulated by iron oxidases (FeOx) and
Fe 2+
reductases (FeR). Fe2+ acts as a co-repressor of the
Fe3+ ferric-uptake regulator (Fur), which is a regulator of the
S transcription of genes that encode iron transporters in
Mms36

R
Fe2+ the cytoplasmic membrane but not of genes with functions
PX

Fe2+

GFDC
e– pool that are specific to magnetosomes. b | Precipitation of
O

iron that is imported into the magnetosome membrane

E T

Fe3+ vesicle occurs in close proximity to the inner leaflet of the

? e–
c such as MamO and MamI. The Fe2+/Fe3+ ratio inside
ΔmamP
ΔmamI
Major facilitator superfamily
(MFS). A family of ubiquitous
membrane proteins that
transports sugars,
oligosaccharides, amino acids,
peptides, drugs, metal ions and
oxyanions, either across an
electrochemical (uniporter) or
a chemiosmotic (symporter
or antiporter) gradient.
Fe3+ Mms48
Δmms6
50 nm
Δmms36
Nature Reviews | Microbiology
MamH and MamZ, which are members of the major
facilitator superfamily (MFS)28. In MSR, deletion of either
mamB or mamM prevented magnetite biomineraliza-
tion, and site-directed mutagenesis of the active (metal
binding) site of MamM strongly impaired biomineraliza-
tion43. Similarly, an MSR double-deletion mutant of both
mamH and mamZ exhibited a substantial decrease in
magnetite biomineralization28, although single-deletion
mutants had much weaker phenotypes, which suggests
that MamH and MamZ, which are partially homologous
to one another, have redundant functions in iron trans-
port into magnetosome membrane vesicles28. Finally,
although the putative iron transporter MagA had pre-
viously been suggested to be involved in magnetosome
biogenesis, genetic experiments in AMB and MSR have
shown that magnetosomes in magA-deletion mutants
resemble those of wild-type cells77.
H+ Mms6
MmsF
magnetosome membrane and is probably mediated by
N
proteins that are located on the magnetosome membrane,
magnetosome membrane vesicles is regulated by MamE,
MamP, MamT and MamX, which are also located on the
magnetosome membrane, and by the activity of respiratory
chains; this ratio ensures that biomineralization produces
the mixed-valence iron mineral magnetite. After
nucleation of the growing magnetite crystal, several
proteins that are located on the magnetosome membrane
regulate the maturation of crystals of defined size and
shape: MamG, MamF, MamD, MamC, MamS, MamR, MamN,
Mms6 and MmsF positively regulate crystal size, whereas
Mms36 and Mms48 negatively regulate crystal size.
c | Transmission electron micrographs of Magnetospirillum
gryphiswaldense MSR‑1 (MSR) deletion mutants that have
defects in redox control and nucleation of magnetite
biomineralization (ΔmamP and ΔmamI) or the maturation
of magnetite crystals (Δmms36 and Δmms6). The
mamP-deletion mutant produced two types of crystals:
large polycrystalline magnetite particles (indicated by the
black arrow) and small, irregular, poorly crystalline
non-magnetic iron oxide particles (such as haematite,
indicated by white arrows). The mamI-deletion mutant
produced only small, irregular, poorly crystalline iron oxide
particles (indicated by white arrows) that were not
magnetic. The mms6‑deletion mutant produced smaller
and fewer magnetite crystals (although in Magnetospirillum
magneticum AMB‑1 (AMB) the shape of crystals was not
affected by the loss of mms6; not shown), whereas the
mms36‑deletion mutant produced crystals that were larger
than those produced by wild-type cells. CM, cytoplasmic
membrane; OM, outer membrane.
Redox control of magnetite biomineralization. As a
mixed-valence iron oxide, the formation of magnet-
ite is only favoured within a narrow redox range (Fe3+
and Fe2+ are present at a ratio of 2/1). Therefore, strong
perturbations of redox conditions, such as high oxygen
levels, growth on highly oxidized carbon sources or
inhibition of respiratory pathways, cause the formation
of smaller and misshapen crystals of magnetite or even

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entirely suppress biomineralization78–83. The expression
of ­magnetosome-associated genes was found to be unaf-
fected by redox conditions84–86 and magnetosome mem-
brane vesicles are present even under redox conditions
that suppress magnetite biomineralization, or indeed
under conditions of iron starvation32,37,39. Therefore,
environ­mental conditions seem to influence the physico-
chemical conditions in the interior of magnetosome
membrane vesicles directly and/or by auxiliary meta-
bolic pathways, rather than through the regulation of
genes with specific functions in magnetosome biogenesis.
Several partially redundant general cellular meta-
bolic pathways have been found to influence the redox
state of iron during anoxic and microoxic growth,
and thus magnetite biomineralization (FIG.  3b). For
example, in MSR, impairment of the reduction of
Fe3+ to Fe2+ through the deletion of two of six cyto-
solic Fe3+ reductases completely inhibited magnetite
bio­mineralization during growth with Fe3+ as the sole
iron source87. In addition to Fe3+ reductases, enzymes
of microoxic or anoxic respiratory chains are involved
in the maintenance of cellular redox conditions. For
example, deletion of the terminal, high affinity cbb3
oxygen reductase, a putative oxygen sensor that might
indirectly regulate redox conditions, impairs magnetite
biomineralization80. Finally, the denitrification pathway
is also important for the maintenance of redox balance.
As such, mutants of MSR that lack essential enzymes of
the denitrification pathway, such as periplasmic nitrate
reductase (Nap), Fe(ii)–nitrite oxidoreductase (NirS)
or nitric oxide reductase (NorCB), have severe defects
in the morphologies, sizes and numbers of magnetite
crystals, as well as in the organization of magnetosome
chains78,79. Transcription of denitrification genes relies
on the oxygen-responsive transcriptional regulator
fumarate and nitrate reduction regulatory protein (Fnr),
the deletion of which produces a severe impairment in
magnetite biomineralization88.
In addition to general metabolic pathways,
­magnetosome-specific factors are thought to contrib-
ute to the regulation of redox conditions, owing to the
presence of certain domains that are associated with
redox functions. For example, deletion of the YedZ-like
iron reductase domain of the putative iron transporter
MamZ produces the same phenotype as deletion of the
whole gene, which suggests that the reduction of iron
is coordinated with its transport 28. MamE, MamP,
MamT and MamX each contain two or three conserved
CXXCH c‑type cytochrome haem-binding motifs18,28,
which are denoted the ‘magnetochrome’ domain,
owing to its restriction to magneto­tactic bacteria89. A
crystal structure of dimeric MamP from the marine
Alphaproteobacterium MO‑1 showed that an acidic
pocket at the dimer interface is enclosed by four mag-
netochrome domains, which suggested that this pocket
may bind to and oxidize iron90. Indeed, in vitro exper-
iments with recombinant MamP from AMB showed
a capability to oxidize Fe2+ and to mineralize mixed-­
valence iron oxides89,91. However, none of the proteins
that are known to have a magnetochrome domain seem
to be essential for magnetite biomineralization, as null
mutants of mamP, mamT or mamX form regular mag-
netite crystals, although the crystals are smaller in size
and number, and, in some mutants, are flanked by tiny,
irregular and poorly crystalline particles24,28,55,91 (FIG. 3c).
Finally, although mamE is an essential gene for magnetite
biomineralization, the magnetochrome domain of MamE
does not seem to be essential for this process29. Thus, pro-
teins with magnetochrome domains may have distinct
and redundant functions in magnetite biomineralization.
Nucleation of magnetite crystals. Nucleation of mag-
netite crystals occurs when, under optimal conditions
(>pH 7 and low redox potential), soluble Fe 2+ and
Fe3+start to form crystalline iron phases. Currently, two
models for the nucleation of magnetite crystals have
been proposed. The first model assumes that magnetite
biomineralization does not involve intermediate min-
eral phases and instead occurs by direct co-­precipitation
of soluble Fe2+ and Fe3+, whereas the second model
involves the formation of precursor mineral phases that
undergo phase transformations to magnetite (reviewed
in detail in REF. 92). Nucleation of magnetite crystals
seems to be highly regulated, as only a single (or some-
times twinned) crystal is formed in each magnetosome
membrane vesicle. Small nascent crystals in the lumen
of magnetosome membrane vesicles have been found
in close proximity to the inner leaflet of the magneto-
some membrane, which suggests that nucleation occurs
at the interface with the magnetosome membrane34,35.
The small, highly abundant magnetosome proteins
Mms5, Mms6, MamC and MamD were initially sug-
gested to function in this nucleation process, owing to
their strong physical association with magnetite crystals
and to the ability of Mms6 and MamC to bind to iron
in vitro. However, AMB and MSR deletion mutants had
only weakly defective magnetite biosynthesis pheno­
types (smaller and sometimes misshapen crystals),
which indicates that these proteins are not responsible
for magnetite nucleation, but that they instead prob-
ably regulate the size and shape of magnetite crystals
at later stages of crystallization20,58,93 (FIG. 3b,c). By con-
trast, the deletion of mamE, mamM or mamO in AMB
and MSR entirely abolished magnetite biomineraliza-
tion but not the formation of magnetosome membrane
vesicles24,43,55,94. MamE, MamM and MamO are thus
candidate regulators of magnetite nucleation, in addi-
tion to, or perhaps as a consequence of, the proposed
functions of these proteins, which, for MamE, include
targeting proteins to the magnetosome membrane and
proteolytic activity (autocleavage and cleavage of MamO
and MamP); for MamM, include iron transport into the
magnetosome and maturation of magnetosome mem-
brane vesicles; and, for MamO, include transition metal
binding and the regulation of MamE-dependent proteo­
lysis29,37,43,95,96. In MSR, deletion mutants that lack mamI,
which encodes a small protein that lacks any discerni-
ble homology to known protein domains, are unable to
biomineralize magnetite but instead produce tiny and
poorly crystalline particles of haematite56, which sug-
gests that MamI may also have a function in magnetite
nucleation (FIG. 3b,c).

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Control of crystal size and number. Magnetite crystals
in mature magnetosomes are highly uniform and have
a narrow size distribution compared with abiotically
synthesized magnetite nanoparticles, which suggests
an exceptional degree of control over crystal growth.
The deletion of many magnetosome-associated genes
causes the production of smaller crystals of variable size
(Supplementary information S1 (table)), which suggests
that these genes encode proteins that have major roles
in the regulation of the crystallization process and size
control. In addition to proteins that are involved in the
earlier steps of bio­mineralization, such as those with
magnetochrome domains (see above), the production of
smaller magnetite crystals has been described in mutants
that lack MamS, MamR, MamN, MamF, Mms5, Mms6,
MamD or MmsF. For the most part, the mechanisms
by which these factors regulate crystal size are not well
understood. Of note, MamN shares similarities with
H+-pumping transporters and has therefore been sug-
gested to export protons that are released by the precipita-
tion of magnetite, and thus maintain the pH of the vesicle
lumen12,56. The magnetite-binding proteins Mms5, Mms6
and MamD, which are specific to magnetotactic bacteria,
are thought to form stabilizing interactions with mag-
netite surfaces during crystal growth59, or, for MamD,
to template the crystal lattice of magnetite92. Consistent
with such roles, the acidic C‑terminal region of Mms6
has been shown to bind to iron in vitro20,61,97,98. In con-
trast to deletion mutants in which the size of magnetite
crystals is smaller, only two genes have been described
as leading to an increase in the size of magnetite crys-
tals when deleted: mms36 and mms48. In these mutants,
magnetite crystals are 10% and 30% larger than in wild-
type cells, respectively 56 (FIG. 3c). Mms48 has a TPR motif
that might mediate protein–protein ­interactions that
influence crystal size56.
Given that a single magnetite crystal is produced for
each magnetosome (see above), the number, as well as the
size, of magnetite crystals seems to be tightly regulated
in magnetosome biogenesis. Indeed, a balance needs to
be struck between the generation of a sufficiently strong
magnetic field sensor and the energetic costs, as well as
the potentially adverse effects of excess magnetosome
biogenesis99. Several single-­deletion mutants have been
described that produce a smaller number of magnetite
crystals compared with wild-type cells, including mamA
and mamP mutants56 (Supplementary information S1
(table)). However, the molecular and structural mech-
anisms that are used to ensure an optimal number of
magnetosomes are not well understood.
Assembly of magnetosome chains
Magnetosomes are aligned into linear chains in which
magnetic moments of individual particles sum up to
maximize the total magnetic response of the cell (FIG. 4a).
Depending on the growth conditions, magnetosome
chains in Magnetospirillum spp. can contain more than
100 magnetosomes. As a string of magnetic dipoles has
an inherent tendency to collapse into flux-closed rings
or agglomerate into irregular clusters, magnetotactic
bacteria use cellular structures to stabilize magnetosome
chains; these structures also anchor the magnetosome
chain so that the whole cell is aligned with magnetic
field lines.
Magnetosome chain assembly results from the coor-
dinated interplay of magnetic interactions, which occur
naturally between magnetite crystals, and active assembly
mechanisms, which are mediated by proteins100. Cryo­
electron tomograms of MSR and AMB revealed the exist-
ence of bundles of cytoskeletal filaments that tra­verse
the cell in close proximity to magnetosome chains34,38
(FIG. 4b,c). These filaments are formed by MamK, which
is an actin-like protein that is mostly restricted to mag-
netotactic bacteria, in which it is highly conserved, and
encoded by a gene in the mamAB operon17,101. In com-
mon with other actin-like proteins, MamK polymerizes
into filaments in vitro102,103 and, depending on ATPase
activity, MamK filaments are dynamic both in vitro and
in vivo30,104,105. Notably, MamK has a distinct intracel-
lular localization to the other actin-like protein that is
present in Magnetospirillum spp.105, MreB, and forms a
distinct branch of the large superfamily of prokaryotic
actin-like proteins, which have diverse functions in cell
­morphology and organization106.
Both MSR and AMB depend on MamK for the
assembly of magnetosome chains, but several differ-
ences have been observed in the mechanisms of chain
assembly between the two model species. In MSR, wild-
type cells have one or two contiguous magnetosome
chains that extend across approximately two-thirds of
the length of the cell and are located at mid-cell, whereas
magnetosome chains in a mamK-deletion mutant,
are shorter, fragmented and ectopically located at an
off-centre position in the cell35,107 (FIG. 4b). The pheno-
type of the mamK mutant suggests that MamK is not
essential for the alignment of magnetosomes into short
chains in MSR, but that it has a crucial role in joining
short chains into a single magnetosome chain and in
ensuring that this chain is positioned at mid-cell. Rather
than simply providing a rigid scaffold for chain assembly,
MamK has been suggested to have an active role in the
process that may involve the generation of mechanical
forces through polymerization and depolymerization,
similarly to other cytomotive cytoskeletal filaments
(such as those formed by ParM)107,108. MamJ, an inter-
acting partner of MamK109,110, has also been shown to
have a role in the assembly of magnetosome chains in
MSR. In mamJ-deletion mutants, magnetosomes detach
from the MamK filament and, once separated from this
scaffold, aggregate by magnetic attraction38,109 (FIG. 4b),
which suggests that MamJ is a connector that attaches
magnetosomes to MamK filaments (FIG. 4a). Consistent
with such a role, MamJ has been shown to bind to the
magnetosome membrane18. The different phenotypes of
the MSR mamJ-deletion and mamK-deletion mutants
indicate the existence of additional as-yet-unidentified
determinants of magnetosome chain formation.
Unlike MSR, wild-type cells of AMB form long mag-
netosome chains that may extend across the entire cell.
However, these chains often include empty vesicles inter-
spersed among vesicles that contain mature crystals, and
can thus seem to be fragmented when cells are visualized

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MamJ

MamK

b
MSR wild-type AMB wild-type

500 nm 500 nm

MSR ΔmamJ MSR ΔmamK

500 nm 500 nm

c MSR wild-type d MSR wild-type

OM
PG
IM
PHB
100 nm

Figure 4 | Magnetosome chain assembly, positioning and segregation. a | Mature magnetosomes (red circles) that
contain cuboctahedral magnetite crystals (dark grey octagons) are aligned into chains through the interaction
Nature Reviews of the
| Microbiology
magnetosome-bound MamJ (yellow star) with the actin-like MamK filament (green dashed line); MamK is also required
for the positioning of the magnetosome chain to the mid-cell. Organization of magnetosomes into chains helps to align
individual magnetic dipoles (white arrows) of single crystals with each other to generate a strong dipole that can ‘sense’
the geomagnetic field. b | Transmission electron micrographs of wild-type Magnetospirillum gryphiswaldense MSR‑1 (MSR),
wild-type Magnetospirillum magneticum AMB‑1 (AMB), a mamJ-deletion mutant of MSR and a mamK-deletion mutant of
MSR. These micrographs show the different magnetosome chain organizations of MSR and AMB, in which MSR has
seemingly contiguous magnetosome chains but ‘gaps’ are visible in the magnetosome chains of AMB, owing to the presence
of ‘empty’ vesicles that do not contain magnetite crystals. The mamJ-deletion and mamK-deletion mutants show the defects
that occur without these key proteins: the loss of mamJ results in the detachment of magnetosomes from MamK filaments
and aggregation of the detached magnetosomes by magnetic attraction, whereas the loss of mamK results in a failure to
translocate to the mid-cell from the pole following cell division and the formation of shorter chains. The difference between
the two phenotypes might be due to the existence of an additional, as-yet-unidentified protein that enables magnetosomes
to remain attached to filaments in the absence of MamK. c | Segmented cryoelectron tomogram of MSR at an early stage of
cell division. Insets show different perspectives of the same tomogram (cell envelope in blue; magnetite crystals in red;
magnetosome membrane in yellow; magnetosome filament in green). d | Section of a cryoelectron tomogram of MSR
wild-type that illustrates the asymmetric indentation of the inner membrane (IM), peptidoglycan (PG) and outer
membrane (OM). PHB, polyhydroxybutyrate granule. Part c and part d are adapted with permission from REF. 107, Wiley.

by conventional transmission electron microscopy 104 MamK-like, that probably also forms filament bundles31,
(FIG. 4b). Similarly to MSR, the filament that is associ- and the filaments formed by each of the two homologues
ated with the magnetosome chain in AMB is formed are expected to interact with each other 104. Deletion of
by MamK; however, AMB has a paralogue of MamK, mamK or mamK-like, or co-deletion of both genes,

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in AMB results in a disorganized magnetosome chain,
although the phenotypes of these mutants were much
weaker than that of the MSR mamK-deletion mutant 104.
Similarly, single deletions of mamJ or its paralogue limJ,
or even the deletion of both genes, cause weaker pheno-
types than the deletion of mamJ in MSR, in that the loss
of these genes produces gaps in magnetosome chains
but the chains are still present. A comparison of the
single-deletion and double-deletion mutants suggests
that MamJ and LimJ have redundant functions, as larger
gaps in magnetosome chains are produced when both
genes are deleted than in the single-deletion mutants30.
An additional interacting partner that has been described,
at least putatively, for MamK in AMB is a methyl-
accepting chemotaxis protein (MCP); this interaction,
together with microscopic analyses, have led to the sug-
gestion that MamK relays the sensing of physical forces
on the magnetosome chain to changes in cell motil-
ity 111,112. However, such a role for MamK has not yet been
confirmed with appropriate mutant strains.
Despite its important role in MSR and AMB, MamJ
does not seem to be a universal mediator of mag-
netosome chain assembly, as it has only been found
in Magnetospirillum spp. and not in other species of
magneto­tactic bacteria. By contrast, genomic surveys
suggest that homologues of MamK are widely present in
magnetotactic bacteria (indeed, are seemingly present
in all magnetotactic bacteria), and in some cases together
with other actin-like proteins (for example, Mad28) that
are speculated to function in the assembly of magneto-
some chains that have more complex architectures than
those found in MSR and AMB1,3,113.
Division and segregation of magnetosome chains. To
ensure proper segregation and equal inheritance of
magnetosomes during cytokinesis, magnetosome chains
are positioned at mid-cell, which is the site of cellular
division107,114 (FIG. 4c). In MSR, this positioning relies on
MamK (see above), and the deletion of mamK results
in mispartitioning of daughter magnetosome chains
to newborn cells107. Segregation of the magnetosome
chains during cell division entails unidirectional inden-
tation, constriction and then asymmetric septation of
the cell wall107, which probably enables the segregation
of daughter magnetosome chains to overcome the sub-
stantial magnetostatic forces between magnetosome
particles107,115 (FIG. 4c,d). Magnetosome segregation was
found to be coordinated with the cell cycle107 and resem-
bles partitioning mechanisms of other bacterial orga-
nelles and macromolecular complexes, such as plasmids,
carboxysomes or cytoplasmic chemoreceptor clusters107.
Importantly, following cell division in MSR, the poles of
daughter cells contain cellular material from mid-cell
of the parent, which means that magnetosome chains
are localized to the poles, rather than mid-cell. In wild-
type MSR cells, these magnetosome chains undergo a
dynamic translocation to mid-cell of the daughter cell
following cytokinesis, but the translocation is abolished
in a mamK mutant 107 (FIGS 1a,4b). According to one
hypothesis, positional information for localization of
the magnetosome chain to the site of cellular division
may be provided by a possible interaction with the cell
division machinery, such as constituents of the divisome.
Indeed, a possible interaction between MamK and FtsZ,
which in most bacteria orchestrates cytokinesis116, was
suggested by the discovery of a hybrid MamK protein
that was fused to an FtsZ homologue in an uncultured
species of magnetotactic bacteria117. As mentioned pre-
viously, a truncated FtsZ homologue known as FtsZm is
encoded in the MAI of MSR, and this protein also local-
izes to the site of cellular division118. However, genetic
studies suggest that FtsZm is not required for cell divi-
sion or magnetosome chain segregation119,120 and that it
may instead be involved in organizing and scaffolding
complexes that are formed by MamX and MamZ28,120.
Magnetosome biogenesis in diverse bacteria
Compared with the rather simple cuboctahedral mag-
netite crystals that are produced by Magnetospirillum
spp., magnetosomes from many magnetotactic bacte-
ria that are found in environmental samples, which are
mostly unculturable, display a spectacular diversity of
composition, shapes, sizes, number and intracellular
organization121 (FIG. 5a–e). Using recent breakthroughs
in the isolation of novel species, targeted meta­genomics
and single-cell genomics, the genomes of numerous cul-
tured and uncultured species of magnetotactic bacteria
have now been analysed, which has revealed the exist-
ence of complex gene clusters that resemble the MAI
of Magnetospirillum spp. in all of the species that have
been studied. These gene clusters are considerably diver-
gent to one another with respect to sequence identity
and gene composition16,122, and the study of how these
genetic differences relate to the diversity of magneto-
some composition, shape, size, number and intracellu-
lar organization will inform future efforts to genetically
engineer magnetosome biogenesis (see below). For
example, homologues of the mamGFDC and mms
operons are only found in those magnetotactic bacteria
that are Alphaproteobacteria, which suggests that these
genes are associated with cuboctahedral or elongated
prismatic magnetite particles, whereas various families
of mad (magnetosome-associated deltaproteo­bacterial)
genes and man (magnetosome specific to Nitrospirae)
genes are specifically present in those magnetobacteria
that are Deltaproteobacteria and/or belong to the uncul-
tured Nitrospirae and Omnitrophica phyla, which sug-
gests that these genes are associated with b­ ullet-shaped
magnetite crystals1–3,113,123. Recently, a genetic screen
in the delta­proteobacterium Desulfovibrio magneticus
confirmed that mutations in several mad genes produce
defects in the biomineralization of magnetite crystals124,
and also identified mam genes, fmp (fewer magnetic
particles) genes and kup, which encodes a potassium
transporter, as genes that may have functions in magne-
tosome biogenesis. Furthermore, a recent study showed
that simultaneous overexpression of almost all mam and
mms gene clusters substantially increased the number
of magnetosomes in MSR125 (FIG. 5f), which indicates
that control of magnetosome number to a large extent
depends on the balanced expression levels of genes that
encode the machinery for magnetosome biogenesis.

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a b c d e

f g Recipient Donor
R. rubrum ‘magneticum’ M. gryphiswaldense

500 nm

50 nm 50 nm

500 nm

+
Magnetosome genes
Expression cassettes

MTB Other bacteria Eukaryotic cells

Strain overproducing magnetosomes R. rubrum

?
? ?
Strain producing modified magnetosomes E. coli Yeast

Nature Reviews | Microbiology

Interestingly, the overexpression of these genes markedly
impaired growth, which suggests that magnetosome over-
production is associated with increased metabolic costs
and/or toxicity. Finally, genetic studies have investigated
why some species of magnetotactic bacteria produce
crystals that are composed of the magnetic iron–sulfur
mineral greigite (Fe3S4), rather than — or in addition to
– ­magnetite. In marine Deltaproteobacteria, distinct var-
iants of MAI-like gene clusters have been associated with
the formation of greigite or magnetite crystals126, and two
related but divergent sets of MAI-like gene clusters were
discovered in Deltaproteobacteria that are able to form
both minerals, which suggests that these bacteria use sep-
arate sets of genes for the biogenesis of greigite magneto-
somes and magnetite magnetosomes2,127. Altogether, the
astonishing diversity of MAI-like gene clusters indicates
that magnetosome biogenesis pathways are somewhat
divergent in different species of magnetotactic bacte-
ria, which accounts for the diversity that is observed in
­m­­agnetosome formation.

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◀ Figure 5 | Diversity and genetic engineering of magnetosomes. Electron micrographs
of magnetosome crystals in various uncultured environmental magnetotactic bacteria,
showing the diversity of magnetosomes that are produced by magnetotactic
bacteria, which include cuboctahedral (part a), elongated prismatic (part b and part c),
tooth-shaped (part d) and bullet-shaped (part e) morphologies. Scale bars, 100 nm.
Transmission electron micrograph of an Magnetospirillum gryphiswaldense MSR‑1 (MSR)
strain in which each magnetosome biogenesis operon had been duplicated. The size
and number of magnetosomes is substantially increased in these cells compared with
wild-type cells (see, for example, FIG. 4a; part f). Cells of a transgenic strain of
Rhodospirillum rubrum that heterologously expresses 30 MSR genes that are associated
with magnetosome biogenesis are ‘magnetized’, which provides proof‑of‑principle
evidence that both magnetosome biogenesis and the magnetotactic trait can be
genetically transferred to non-magnetotactic bacteria. Transmission electron
micrograph of a mixed culture showed that the ‘magnetized’ strain of R. rubrum has the
characteristic magnetosome organization of MSR (part g), and that the ‘magnetization’
of the strain could be demonstrated by accumulation of bacterial cells near the poles of a
magnet (part h). By varying the copy number and composition of magnetosome gene
clusters, genome engineering of various cultured and uncultured magnetotactic
bacteria (MTB) could be used to produce magnetosomes in increased numbers and/or of
modified sizes, morphologies, mineral composition, intracellular alignment and
magnetic properties. Expression cassettes could also be transferred to non-magnetotactic
bacteria (for example, R. rubrum or Escherichia coli), or possibly even to yeast or other
eukaryotic cells, to endow them with genetically encoded magnetic properties (part i).
Parts a–e are adapted with permission from REF. 169, Horizon Scientific Press. The image
in part f is from Appl. Environ. Microbiol., 2016, 82, 3032–3041, http://dx.doi.org/10.1128/
AEM.03860-15 and amended with permission from American Society for Microbiology.
Part g and part h are from REF. 129, Nature Publishing Group.
Despite the considerable diversity in genes that are
associated with magnetosome biogenesis, only a small
set of nine genes (mamABEKMOPQI) are present in
all genomes of magnetotactic bacteria that have been
studied to date (mamL is additionally present in all
magnetotactic bacteria that produce only magnetite
— as opposed to greigite — crystals1), which suggests
that only the core functions of correct biogenesis and
assembly of the magnetosome membrane, Fe2+ uptake,
magnetite biomineralization and magnetosome chain
assembly are mediated by universal mechanisms in all
magnetotactic bacteria. Interestingly, the core genes
that are associated with magnetotactic bacteria are not
present in eukaryotic organisms that have been shown
to biomineralize magnetite (these organisms include
migratory fish and birds), which suggests that magne-
tosome biogenesis in bacteria is evolutionary unrelated
to eukaryotic pathways for magnetite biomineralization,
contrary to earlier speculations128.
‘Magnetizing’ non-magnetotactic bacteria. A major
objective of studying magnetosome biogenesis is
to inform the development of applications that use
magnetism in biotechnology (BOX 2). However, most
­magnetotactic bacteria are recalcitrant to cultivation in
the laboratory, fastidious to grow and/or cumbersome to
analyse by genetic methods, and so much of the consid-
erable diversity of magnetotactic bacteria has not been
explored in the laboratory. The development of a model
in which the entire magnetosome pathway is expressed in
a bacterium that is more facile to study may therefore
provide a useful alternative to laboratory investigations
of naturally magnetotactic bacteria. However, such
a model might not be trivial to develop, owing to the
genetic complexity, and our incomplete knowledge, of
magnetosome biogenesis. In a recent study, the entire
pathway of magnetosome biogenesis in MSR (30 MAI
genes, which were assembled into several expression
cassettes) was transferred to a non-magnetotactic bac-
terium (the photosynthetic bacterium Rhodospirillum
rubrum)129, which represented a proof‑of‑principle
demonstration that magnetosomes can be fully recon-
stituted in non-magnetotactic bacteria, despite the struc-
tural complexity of these organelles. Expression of the
transferred genes resulted in a ‘magnetized’ R. rubrum
(FIG. 5 g,h), with the bacterium acquiring the capability for
magnetosome biogenesis, the formation of well-ordered
magnetosome chains and accumulation near the pole
of a magnet. Genetic studies of ‘magnetized’ R. rubrum
showed that the deletion of individual MAI genes pro-
duced similar phenotypes to those observed when the
same genes were deleted in MSR, which confirmed that
heterologous expression can be used to genetically dis-
sect magnetosome biogenesis in bacteria that are more
amenable to laboratory study than naturally magneto-
tactic species. The ability of R. rubrum to become mag-
netotactic following the transfer of only a relatively small
number of genes also provides the first experimental
evidence that the dissemination of the magnetotactic
trait across genus boundaries can occur as the result of
horizontal gene transfer events that are likely to occur
under natural conditions. Whether such a mechanism is
responsible for the presence of magnetosome biogenesis
in different genera, or whether the distribution of this
trait is instead explained by vertical inheritance from a
common magnetotactic ancestor, has been the subject
of debate3,124,130,131.
Outlook
In the past decade, remarkable progress has been made
in understanding the molecular genetics and cell biol-
ogy of magnetotactic bacteria. Using MSR and AMB
as genetically tractable models, much has been learned
about the individual roles of MAI genes in magneto-
some biogenesis. For example, studies of the core genes
mamA, mamB, mamE, mamK, mamM, mamO, mamP,
mamQ, mamI and mamL, which seem to be shared by
all magnetotactic bacteria that produce magnetite crys-
tals, have shown that mamA, mamB, mamE, mamL and
mamQ are required for the correct biogenesis and assem-
bly of the magnetosome membrane; mamB and mamM
are required for Fe2+ uptake; mamE, mamP and mamI
are required for the nucleation, partial oxidation and
crystallization of magnetite; and mamK is required for
the assembly and organization of magnetosome chains.
However, despite the exciting developments of recent
years, many unsolved aspects of magnetosome bio­
genesis remain. For example, the mechanisms that gov-
ern the formation of magnetosome membrane vesicles
are still not fully understood and how proteins are tar-
geted to the magnetosome membrane has not yet been
established. With regards to biomineralization, what
are the mechanisms that underlie the crystallization of a
single, stoichiometrically pure magnetite particle inside
each magnetosome membrane vesicle? To answer this

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Magnetic heating Box 2 | Biotechnological applications of magnetosomes

The process of heat release by
magnetic nanoparticles that
are exposed to alternating
magnetic fields.
Colloidal stability
The tendency of particles to
remain suspended in solution.
Purified bacterial magnetosomes represent magnetic nanoparticles with exceptionally well-defined characteristics,
owing to the precise control that is exerted during all stages of biogenesis, and several unprecedented properties, such
as high crystallinity, strong magnetization, and a uniform distribution of shape and size that cannot be replicated by
synthesis using abiotic processes. The combination of these features, the potential for genetic engineering and
biocompatibility have made magnetosomes highly attractive for use in many bioremediation, biomedical and bionano-
technological applications146–148. For example, magnetosomes targeted to cancerous tissues are highly promising for
use in the hyperthermic treatment of tumours149–151, owing to an exceptional magnetic heating capability152–156.
The defined composition of the proteins that are associated with the magnetosome membrane provides an excellent
target for chemical modification157,158, which has been used for the magnetosome-specific display of fluorophores, or the
genetic construction of fusion between heterologous proteins and the abundant MamC48,50,159. The construction of such
fusion proteins enables heterologous proteins to be targeted with high specificity to the magnetosome surface, which
can be used as a platform for the assembly of multimeric enzyme complexes146,160–164 or the creation of intracellular
nanotraps composed of antibody fragments (‘nanobodies’) that are able to sequester proteins and organelles in living
bacteria, either to manipulate protein function or to construct synthetic cellular structures161,165. Magnetosomes that are
coated with a heterologously expressed protein can be purified and coated with shells of silica or zinc oxide in vitro by
chemical encapsulation166, which improves colloidal stability, enhances resistance to proteases and detergents and is
expected to decrease immunogenicity, thus optimizing magnetosomes for in vivo applications166.
Although the biotechnological potential of bacterial magnetosomes has been demonstrated in the laboratory, no
application has yet been developed for commercial use, owing to the poor scalability and prohibitive cost of magnetosome
production using naturally magnetotactic bacteria. However, the possibility of the genetic transfer of magnetosome
biogenesis to bacterial species that are more amenable to laboratory cultivation and genetic manipulation129, together
with improvements in the scalability of culturing of naturally magnetotactic bacteria167,168, may overcome obstacles to
commercialization and thus spur the development of new biotechnological applications for magnetosomes.

key outstanding question, the molecular interactions at
the organic–inorganic interface will need to be explored
in more detail. Finally, the different genetic pathways
that are associated with the diverse magnetosomes that
have been observed in various cultivated bacteria will
need to be elucidated, including for the many new strains
that have recently been isolated in pure culture127,132–134,
which now await genetic analyses. Such genetic inves-
tigations may use a similar strategy to that recently
applied to the hitherto intractable D. magneticus 125, in
which chemical mutagenesis followed by whole-genome
sequencing identified several novel genes involved in
magnetosome biosynthesis. Alternatively, the functions
of single genes or even entire clusters from uncultured
magnetotactic bacteria could be elucidated by expressing
them in various mutant strains of genetically tractable
Magnetospirillum species.
Biotechnological applications of magnetosome bio-
genesis also offer many exciting future directions for
the field. In particular, the functional transfer of the
entire magnetosome biogenesis machinery to a hith-
erto non-magnetic bacterium means that the genetic
‘magnetization’ of non-magnetic organisms can now
be viewed as feasible. For example, the generation of
a magnetized Escherichia coli strain would enable the
genetic study of magnetosome biogenesis in a fac-
ile bacterium for laboratory investigation, and would
also provide a model system for the modification and
engineering of magnetosome pathways using synthetic
biology approaches. Indeed, one interesting engineering
strategy would be to combine genes from bacteria that
produce diverse magnetosomes; such a ‘mix-and-match’
reconfiguration of the magnetosome biogenesis pathway
might enable the production of designer nano­particles
with finely tuned magnetic properties that could be of
value in numerous nanotechnological and biomedical
applications (BOX 2). However, the transfer of complex,
lineage-specific magnetosome biogenesis pathways into
distantly related, non-magnetotactic species is expected
to require adaptations of the implanted gene cassettes
and of the encoded pathway, as well as modifications to
the host, for optimal function (FIG. 5i). Nevertheless, with
increasing knowledge and technical advances, the gener-
ation of a synthetic toolkit for the genetic magnetization
of both unicellular and multi­cellular organisms can now
be envisioned135. Such a toolkit would enable the intro-
duction of tailored magnetic nanostructures that could
be used as intracellular magnetic labels and tracers or, in
a process known as magnetogenetics, that enable cells to
be analysed using magnetic stimulation136,137.

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An article that describes the discovery of a novel
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Acknowledgements
The authors thank their co-workers and colleagues for helpful
discussions. Work in the Schüler laboratory is supported by
grants from the Human Frontier Science Program (HFSP), the
Deutsche Forschungsgemeinschaft and the European
Research Council (ERC).
Competing interests statement
The authors declare no competing interests.
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