The Ecology and Biotechnology of Sulphate-Reducing Bacteria: Gerard Muyzer and Alfons J. M. Stams

REVIEWS
The ecology and biotechnology of

sulphate-reducing bacteria
Gerard Muyzer* and Alfons J. M. Stams‡
Abstract | Sulphate-reducing bacteria (SRB) are anaerobic microorganisms that use sulphate
as a terminal electron acceptor in, for example, the degradation of organic compounds. They
are ubiquitous in anoxic habitats, where they have an important role in both the sulphur and
carbon cycles. SRB can cause a serious problem for industries, such as the offshore oil
industry, because of the production of sulphide, which is highly reactive, corrosive and toxic.
However, these organisms can also be beneficial by removing sulphate and heavy metals
from waste streams. Although SRB have been studied for more than a century, it is only with
the recent emergence of new molecular biological and genomic techniques that we have
begun to obtain detailed information on their way of life.
Chemolithotropic
Sulphur is among the most abundant elements on the organic carbon mineralization in marine sediments1,
Metabolism of an organism Earth. It is mainly present as pyrite (FeS2) or gypsum which indicates the importance of sulphate reducers
that obtains energy from (CaSO4) in rocks and sediments and as sulphate in in both the sulphur and carbon cycles and, conse-
inorganic compounds and seawater. The sulphur cycle (FIG. 1) is complex, because quently, why SRB have been studied extensively 2. In
carbon from carbon dioxide.
sulphur has a broad range of oxidation states, from –2 this Review, we provide an overview of the diversity,
(completely reduced) to +6 (completely oxidized), and physiology and distribution of SRB and their applica-
can be transformed both chemically and biologically. tions in environmental biotechnology for the removal
In addition, the sulphur cycle is closely linked to other of heavy metals and sulphur compounds from waste
element cycles, such as the carbon and nitrogen cycles. water and flue gas.
Microorganisms play an important part in sulphur
transformations (FIG. 2). Sulphate is taken up as a nutri- Physiology of SRB
ent and reduced to sulphide, which is then incorporated Electron-donor metabolism. Until the early 1980s,
into sulphur-containing amino acids and enzymes. it was thought that sulphate reducers played only a
Oxidation and reduction reactions for the generation of minor part in the carbon cycle. The Desulfovibrio and
metabolic energy are also important, such as sulphide Desulfotomaculum species that were known at that
oxidation by chemolithotropic sulphur bacteria and dis- time used hydrogen and a number of organic com-
similatory sulphate reduction by sulphate-reducing pounds, such as ethanol, formate, lactate, pyruvate,
bacteria (SRB). Because members of the Bacteria and malate and succinate, for growth. Typically, carbon
Archaea can use sulphate as a terminal electron accep- compounds are incompletely oxidized to acetate by
tor, some researchers use the term sulphate-reducing these SRB. However, through the research of Fritz
*Department of
prokaryotes or sulphate-reducing microorganisms. In Widdel at the University of Göttingen, Germany, it
Biotechnology, Delft this Review, however, we use the term SRB to refer to became clear that, particularly in marine sediments,
University of Technology, members of both domains. SRB are the main players in anaerobic carbon cycling.
Julianalaan 67, 2628 BC SRB are anaerobic microorganisms that are wide- Widdel3 isolated and characterized a large number of
Delft, The Netherlands.
spread in anoxic habitats, where they use sulphate novel sulphate reducers that had the ability to grow
‡
Laboratory of Microbiology,
Wageningen University, as a terminal electron acceptor for the degradation on short-chain fatty acids (including acetate), long-
Dreijenplein 10, 6703 HB, of organic compounds, resulting in the production of chain fatty acids and aromatic compounds, such as
Wageningen, sulphide. Subsequently, the sulphide can be oxidized benzoate and phenol. Currently, sulphate reducers can
The Netherlands. under oxic conditions by chemolithotrophic sulphur be divided into two main groups: those that degrade
Correspondence to G.M.
e‑mail: [email protected]
bacteria or under anoxic conditions by phototrophic organic compounds incompletely to acetate and those
doi:10.1038/nrmicro1892 sulphur bacteria. It has been estimated that sulphate that degrade organic compounds completely to car-
Published online 7 May 2008 reduction can account for more than 50% of the bon dioxide. Sulphate reducers that degrade organic
nature reviews | microbiology volume 6 | june 2008 | 441

© 2008 Nature Publishing Group
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Sulphur oxides
in atmosphere
Dry deposition of Acidic precipitation

Sulphur dioxide from sulphate and sulphur (rain and snow)
combustion of fossil dioxide
fuels and sulphide
metal ores
Sulphur dioxide and
sulphate from volcanoes
and hot springs
Sulphate in
Plant uptake water
Mining
Animals
Decomposition Sulphates
in soil Organic Sulphate
Reduced sulphur and other deposition reduction
(hydrogen sulphide) processing
Gypsum and pyrite in Microorganisms

soils and sediments
Sulphur in Deposition of
fossil fuels sulphides to sediment
NatureFeS
Figure 1 | The sulphur cycle. The largest sulphur reservoirs on the Earth are iron sulphides (pyrite; Reviews Microbiology
) and| gypsum
2
(CaSO4) in sediments and rocks (7,800 x 1018 g sulphur) and sulphate in seawater (1,280 x 1018 g sulphur). Sulphur, which is a
necessary element for life, is taken up as sulphate by microorganisms and plants, and subsequently by animals.
Decomposition of dead organisms in the absence of oxygen releases the sulphur again as hydrogen sulphide. The
combustion of fossil fuels and emission of volcanic fumes releases sulphur dioxide into the atmosphere, where it reacts
with water, thereby forming sulphuric acid and resulting in acid rain. Microorganisms play an important part in the
recycling of these sulphur compounds.
compounds completely to carbon dioxide commonly The anaerobic oxidation of methane can be cou-
also use acetate as a growth substrate and two dif- pled to sulphate reduction, as proposed by Reeburgh27
ferent pathways for acetate oxidation are employed, in 1976. Much research has been done to unravel the
a modified citric acid cycle, as used by Desulfobacter microbiology of sulphate-dependent methane oxidation.
postgatei4, and the acetyl-CoA pathway, as used by, for There is solid evidence that this process is carried out
Citric acid cycle example, Desulfobacterium, Desulfotomaculum and by syntrophic communities of archaea, which perform
A cyclic series of reactions that Desulfococcus species5 and Desulfobacca acetoxidans6. reverse methanogenesis, and SRB that oxidize the inter-
result in the conversion of A huge range of novel sulphate reducers have been mediates formed by the archaea28–32, the identities of
acetate to carbon dioxide and
NADH.
described over the past 25 years that have the ability to which are still unknown. Initially, one intermediate was
grow on various different substrates, including sugars7,8, thought to be hydrogen28; however, research by Nauhaus
Acetyl-CoA pathway amino acids9,10 and one-carbon compounds, such as et al. 33 excluded hydrogen, formate, methanol and
A pathway of autotrophic methanol11,12, carbon monoxide13,14 and methanethiol15. acetate as intermediates. The option of methyl sulphide
carbon dioxide fixation and
SRB can also grow by the dismutation of thiosulphate, as an intermediate has also been proposed34. Typically,
acetate oxidation in obligate
anaerobes.
sulphite and sulphur, which results in the formation of archaea are phylogenetically most closely related to
sulphate and sulphide16,17. In addition to benzoate and the Methanosarcina genus and the sulphate reducers
Dismutation phenol, aromatic hydrocarbons (for example, toluene to the Desulfosarcina–Desulfococcus, Desulfobulbus or
The splitting of a chemical and ethylbenzene) are also degraded by a number of Desulfobacter genera29,35–37. However, successful attempts
compound into two new
compounds, one that is more
SRB18–20. Recently, SRB that can grow on long-chain to enrich these SRB from methane-oxidizing sediments
oxidized and one that is more alkanes21–24, alkenes25 and short-chain alkanes26 have also have not yet been reported.
reduced than the original been described. Typically, polymeric organic compounds,
compound. such as starch, cellulose, proteins, nucleic acids (DNA and Electron-acceptor metabolism. Sulphate reducers use
RNA) and fats are not direct substrates for SRB. Therefore, sulphate as the terminal electron acceptor for growth.
Syntrophic
Growth of two or more
in nature, SRB are dependent on other microorganisms However, from a chemical viewpoint, sulphate is an
organisms that depend on that degrade these polymeric substrates and ferment them unfavourable electron acceptor for microorganisms.
each other for their growth. to products that are substrates for SRB (FIG. 3a). The E 0ʹof the redox couple sulphate–sulphite is –516 mV,
442 | june 2008 | volume 6 www.nature.com/reviews/micro

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As many SRB are able to grow on hydrogen and

So
sulphate as sole energy substrates, it is clear that sul-
phate reduction results in electron-transport phos-
phorylation. More than two ATP molecules need to
Ch
be synthesized by electron-transport phosphorylation
io
em
at
id
ol
ox
to compensate for the loss of ATP that is necessary
it
ho
ic Oxic
ph
tro
for sulphate activation. By comparing yields of a
tro
ph
ho
ic
Desulfovibrio strain grown with hydrogen and sul-
ox
it
ol
id
em
at
phate or hydrogen and thiosulphate, a net yield of one
io
Ch
SH groups
n
Des
ti on of proteins ulph ATP molecule per sulphate reduced was proposed
s imila uryl
atio
te a
s
n by Badziong and Thauer 40. Taking into account the
Su lpha DMSO DMS
energy costs for the uptake of sulphate, the net yield
SO42– reduction
would therefore be one-third of an ATP molecule
SO42– H2S or one-quarter of an ATP molecule per sulphate
Sulp reduced41. When a Desulfovibrio strain is growing on
h ate DMS
as DMSO tion lactate, substrate-level phosphorylation also occurs. The
sim ryla
ilati ulphu
on
Des observation that hydrogen is formed when SRB are
SH groups growing on lactate plus sulphate led Odom and Peck42
n
Su
io
Ph
of proteins
io to propose the hydrogen-cycling model. In this model,
at
lp
ot
at
id
hu
ot
t i o tion
ox
rd
ro
lactate is first converted to acetate, carbon dioxide and
ic
r
p
isp
po
ph
hi
ro
ca
ro
ro
n
hydrogen; the hydrogen diffuses out of the cell

po
isp
ot
nd
Anoxic
rti
uc
rd
ith
ch
on
and is used as an electron donor for sulphate reduction.

d
ol
hu
em
re
at
em
So
lp
io
ol
Su
ch
This model, even today, is controversial, but has never

n
ith
nd
ot
ca
ro
been refuted or confirmed convincingly. Hydrogen

p
hi
hi
p
co
ro
formation during growth on lactate might reflect the

ot
xid
ot
at
Ph
high investment of ATP to transport sulphate across

io
n
So
the cytoplasmic membrane and to activate sulphate to
APS. After a period of starvation, the ATP levels in
Figure 2 | Sulphur transformations. Sulphate-reducing Nature Reviews
bacteria have |aMicrobiology
key role in
the sulphur cycle. They use sulphate (SO42–) as a terminal electron acceptor in the
the cell are probably low. Sulphate-independent lactate
degradation of organic matter, which results in the production of hydrogen degradation might be a way to produce the ATP that is
sulphide (H2S). Subsequently, the sulphide can be oxidized aerobically by needed to initiate sulphate metabolism.
chemolithotrophic sulphur-oxidizing bacteria (for example, Thiobacillus or Although named after their ability to use sulphate
Beggiatoa spp.) or anaerobically by phototrophic sulphur bacteria (for example, as a terminal electron acceptor, sulphate reducers can
Chlorobium spp.) to elemental sulphur (S°) and SO42–. Other transformations, which use many other electron acceptors for growth and
are carried out by specialized groups of microorganisms, result in sulphur can ferment substrates in the absence of inorganic electron
reduction (for example, Desulfuromonas spp.) and sulphur disproportionation acceptors. Therefore, the occurrence of high numbers of
(Desulfovibrio sulfodismutans). Organic sulphur compounds, such as SRB in an environment does not necessarily reflect the
dimethylsulphoxide (DMSO) can be transformed into dimethylsulphide (DMS) and
occurrence of sulphate reduction in that environment,
vice versa by several groups of microorganisms. SH, sulfhydryl. Figure modified,
with permission, from REF. 148 (2006) Pearson Education.
but in many recent publications this link is made too
easily. Sulphate reducers can reduce other sulphur com-
pounds (thiosulphate, sulphite and sulphur) to sulphide
which is too negative to allow reduction by the intrac- or can reduce nitrate and nitrite to ammonium43–46. Even
ellular electron mediators ferredoxin or NADH (E 0 ʹ of oxygen respiration is performed by sulphate reducers
–398 mV and –314 mV, respectively) that are present in (BOX 1). Other compounds that are electron acceptors
sulphate reducers. Therefore, before reduction, sulphate for some SRB include iron (Fe(III))47,48, uranyl (U(VI))49,
is activated by an ATP sulphurylase, resulting in the pertechnetate (Tc(VII))50, selenate (Se(VI))51, chromate
formation of adenosine-phosphosulphate (APS) and (Cr(VI))52 and arsenate (As(VI))53. However, not all of
pyrophosphate, which is hydrolysed by pyrophosphatase these reduction processes are coupled to growth.
to 2‑phosphate. The E 0ʹ of the redox couple APS–sulphite Organic compounds can also be used as terminal
plus AMP is –60 mV, which allows the reduction of electron acceptors for growth. Fumarate is used as
APS with reduced ferredoxin or NADH. AMP, which is an electron acceptor by many SRB. Some marine SRB
formed by the reduction of APS, is converted by ATP- use dimethylsulphoxide as an electron acceptor 54.
dependent adenylate kinase into two molecules of ADP. Additionally, sulphonates can act as electron accep-
Thus, the activation of sulphate occurs at the expense of tors for SRB55. Desulfomonile tiedjei has been isolated
two ATP molecules. Sulphite is further reduced to sul- from a methanogenic microbial community that
Substrate-level phide; the E 0 ʹ of the redox couple sulphite–sulphide is mineralizes 3‑chlorobenzoate. In this community,
phosphorylation –116 mV, but how sulphite is reduced to sulphide is not D. tiedjei grows by the reductive conversion of mono-
Synthesis of high-energy
yet clear. A pathway through trithionate and thiosulphate chlorobenzoate to benzoate, with hydrogen formed by
phosphate bonds through the
reaction of inorganic
would allow a reduction in three two-electron reduction benzoate-degrading bacteria56. Interestingly, D. tiedjei
phosphate with an activated steps, but a reduction in one six-electron reduction step was only identified as being a member of the SRB after
organic substrate. still cannot be excluded38,39. it had been isolated57.

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a b
Organic macromolecules Organic macromolecules
(proteins, polysaccharides and lipids) (proteins, polysaccharides and lipids)
Hydrolysis Hydrolysis
Monomers (amino acids, sugars and Monomers (amino acids, sugars and
long-chain fatty acids) long-chain fatty acids)
Fermentation Fermentation
SO42–
Reduced compounds Sulphate Reduced compounds
(lactate, butyrate and propionate) reduction (lactate, butyrate and propionate)
S2–
SO42–
Sulphate Fermentation
reduction
S2–
SO42–
Acetogenesis
H2 Sulphate Acetate H2 and CO2 Acetate
reduction
S2– SO42–
Sulphate Methanogenesis
reduction
S2–
CO2 CH4 and CO2
Figure 3 | The sequential pattern of microbial degradation of complex organic matter in anoxic environments in
the presence and absence of sulphate. Macromolecules, such as proteins, polysaccharides and lipids are hydrolysed by
Nature Reviews | Microbiology
hydrolytic bacteria. Subsequently, the monomers — amino acids, sugars and fatty acids — are fermented by fermentative
bacteria into a range of fermentation products, such as acetate, propionate, butyrate, lactate and hydrogen. In the
presence of sulphate (a), sulphate-reducing bacteria consume these fermentation products. However, in the absence of
sulphate (b), hydrogen and acetate — the acetate having been produced directly by fermentation or indirectly by
acetogenesis — are consumed by the methanogens.
In freshwater environments, which are low in sul- in which one sulphate reducer oxidizes the lactate and
phate, SRB have an important role in the fermentation another uses the hydrogen for sulphate reduction.
and anaerobic oxidation of organic compounds. Many Desulfobulbus species grow on propionate and sul-
Desulfovibrio and Desulfomicrobium species grow by phate, but unlike Syntrophobacter species they cannot
fermenting pyruvate to form acetate, carbon dioxide oxidize propionate to acetate in co-culture with metha-
and hydrogen as products. They are also able to oxidize nogens. However, in the absence of sulphate, they can
lactate and ethanol to acetate, but only when hydrogen ferment lactate and ethanol (plus carbon dioxide) to
is efficiently removed by hydrogen-consuming metha- acetate and propionate.
nogens. This syntrophic growth of sulphate reducers Fermentative and acetogenic growth of SRB might
with methanogens was first demonstrated by Bryant not only explain why they are present in high numbers
and colleagues58. Furthermore, sulphate reducers were in anaerobic environments that are low in sulphate, but
the dominant acetogenic bacteria in a methanogenic also why the addition of sulphate to sulphate-depleted
reactor that was used to treat whey59. sediments results in instantaneous sulphate reduction.
Syntrophobacter species are a special group of sulphate
reducers60. They can grow on propionate and sulphate, Competition with methanogens and acetogens
but were isolated as bacteria that grow by converting In anaerobic environments that have a low redox
propionate to acetate, carbon dioxide and hydrogen potential, SRB compete with other anaerobes, includ-
in a co-culture with hydrogen-utilizing methanogens. ing fermentative bacteria, proton-reducing acetogenic
Similarly, sulphate-dependent or syntrophic growth was bacteria, homoacetogens and methanogens, for the
found for Desulfotomaculum thermobenzoicum subsp. available common substrates. Some important conver-
thermosyntrophicum 61. Syntrophobacter wolinii was sions are listed in TABLE 1. The presence of sulphate is
obtained in a defined co-culture with a Desulfovibrio crucial in this competition. The degradation of organic
species62. S. wolinii is a sulphate reducer that, in the matter in sulphate-reducing environments (FIG. 3a) is
presence of a hydrogen-utilizing sulphate reducer, sup- different from the degradation in methanogenic envi-
Homoacetogen presses sulphate reduction and grows as an acetogen63,64. ronments60 (FIG. 3b). In contrast to sulphate reducers,
A bacterium that produces One can speculate if this is not a more common property methanogens use a limited number of substrates for
acetate as the sole product
from sugar fermentation or
among SRB. An alternative way to interpret the hydro- growth. Quantitatively, hydrogen, carbon dioxide
from hydrogen and carbon gen-cycling model of Odom and Peck42 is that in mixed and acetate are the most important and best-known
dioxide. cultures with SRB syntrophic degradation takes place, substrates for methanogens. Thus far, no methanogens

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Box 1 | Sulphate reducers and oxygen

In mixed microbial communities, sulphate-reducing
bacteria (SRB) are present in the vicinity of microorganisms
that consume oxygen, which creates conditions in which
strict anaerobic bacteria can thrive. This finding was made
more than 100 years ago by the Dutch microbiologist and
founder of the Delft School of Microbiology Martinus
Beijerinck, when he described Spirillum desulfuricans137,
which was later reclassified as Desulfovibrio desulfuricans
(see the figure). At that time, the environmental conditions
and optimal medium composition for SRB were unknown,
but by adding aerobic bacteria Beijerinck obtained better
and more reproducible growth of the SRB. The view that
sulphate reducers are strictly anaerobic, which can still be
found in recent publications, started to change with the
demonstration of the occurrence of sulphate reduction in
oxic environments138. Much research has been done since
then to obtain some insights into the oxygen response of
SRB139. Some strains of sulphate reducers are irreversibly
inactivated by low oxygen concentrations, whereas others
survive aeration, even though sulphate reduction is
suppressed by oxygen140,141. Dilling and Cypionka142
described SRB that respire using oxygen and can even
couple aerobic respiration to ATP formation. Desulfovibrio
oxyclinae, which was isolated from the upper layer of a
hypersaline microbial mat, showed oxygen-dependent
growth, as indicated by higher growth yields after
exposure to oxygen143. The figure is a reproduction of
Vibrio desulfuricans, which was painted by Henriëtte
Beijerinck, the sister of Martinus Beijerinck, and is
reproduced courtesy of the Beijerinck Museum.
have been described that grow on organic acids, such sulphate reducers grow much faster than syntrophic
as lactate, propionate and butyrate, which are com- propionate- and butyrate-degrading methanogenic
mon substrates for sulphate reducers. Consequently, or sulphate-reducing communities, which gives these
these compounds are degraded by bacteria to form sulphate reducers a competitive advantage.
products that are the substrates for methanogens From an ecological viewpoint, it is interesting
(FIG. 3b). Typically, these conversions are carried out to understand how sulphate reducers interact with
by syntrophic communities of acetogenic bacteria and methanogenic communities when the sulphate that
methanogenic archaea. is available is insufficient for complete oxidation of
In the presence of an excess of sulphate, sulphate organic compounds. Under these conditions, SRB
reducers compete with methanogens for the common will compete with each other for the available sulphate.
substrates hydrogen and acetate and with syntrophic Unfortunately, only a few studies have addressed the
methanogenic communities65. Owing to the higher competition between sulphate reducers for sulphate.
affinity and lower threshold values for hydrogen, Laanbroek et al.71 found that Desulfovibrio spp. had the
hydrogen-utilizing methanogens and homoacetogens highest affinity for sulphate followed by Desulfobulbus
are easily and rapidly out-competed by hydrogen- spp. and Desulfobacter spp. This suggests that under
utilizing SRB. However, many SRB have a requirement sulphate limitation sulphate reducers use hydrogen,
for acetate as a carbon source and, therefore, when ace- lactate and ethanol as substrates, but not propionate
tate is not provided, sulphate reducers will coexist with and acetate. It is likely that under sulphate-limited
homoacetogens66,67. Acetate-utilizing sulphate reducers conditions, syntrophic communities have a role in the
also out-compete acetoclastic methanogens68,69. However, degradation of organic acids, whereby the hydrogen-
this competition is not so clear-cut as for hydrogen. utilizing methanogens are replaced by hydrogen-utilizing
In experiments in which sulphate was added to a fully sulphate reducers.
methanogenic anaerobic bioreactor, it took years before
the acetotrophic Methanosaeta species were out-com- Diversity and activity of SRB
peted by sulphate reducers70. The sulphate reducer Different techniques have been used to detect SRB
Acetoclastic methanogen that became dominant was Desulfobacca acetoxidans, and study their diversity and activity. One of the old-
A methanogen that uses
acetate as a substrate to
a bacterium that is specialized in growth on acetate 6 est techniques that has been used in this context is
produce methane and carbon and has only slightly better growth kinetics than cultivation. Although successful, this technique has
dioxide. Methanosaeta spp. Propionate and butyrate-degrading limitations, as only a small percentage of bacteria

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Table 1 | Sulphate-reducing, methanogenic and acetogenic reactions the sulphate-reduction pathway, such as dsrAB, which
encodes the dissimilatory sulphite reductase77, or aprBA,
Equation ∆Goʹ which encodes the dissimilatory adenosine‑5′-phospho-
(kJ/reaction)*
sulphate reductase78. Cloning or denaturing gradient gel
Sulphate-reducing reactions electrophoresis of PCR-amplified 16S rRNA79,80, dsr81–83
4 H2 + SO42– + H+ → HS– + 4 H2O –151.9 or aprA84 gene fragments has been used to determine
Acetate– + SO42– → 2 HCO3– + HS– –47.6 the diversity of SRB in many different habitats. Recently,
a DNA microarray, the SRP-PhyloChip85, has been used
Propionate + 0.75 SO
– 2–
→ Acetate + HCO + 0.75 HS + 0.25 H
– – – +
–37.7
4 3 to detect SRB in natural samples, such as acidic fen
Butyrate– + 0.5 SO42– → 2 Acetate– + 0.5 HS– + 0.5 H+ –27.8 soils86. However, these methods have the disadvantage
Lactate + 0.5 SO
–
4
2–
→ Acetate + HCO + 0.5 HS
–
3
– –
–80.2 that they provide little or no information on the number
Acetogenic reactions of SRB cells that are present.
Quantitative real-time PCR is a highly sensitive
Propionate– + 3 H2O → Acetate– + HCO3– + H+ + 3 H2 +76.1 technique that can be used to quantify the number
Butyrate– + 2 H2O → 2 Acetate– + H+ + 2 H2 +48.3 of SRB, and has been used, for example, to determine
Lactate + 2 H2O → Acetate + HCO + H + 2 H2
– – – +
–4.2 the number of SRB in rice field soils87,88, soda lakes89
3
and industrial waste water90. Moreover, this technique
Methanogenic reactions
can also be used to study the expression of functional
4 H2 + HCO3– + H+ → CH4 + 3 H2O –135.6 genes, such as dsrAB91. Another technique that can be
Acetate– + H2O → CH4 + HCO3– –31.0 used to quantify the number of SRB is fluorescence
Homoacetogenic reactions in situ hybridization (FISH), which also allows their
spatial distribution to be visualized81,92. Many differ-
4 H2 + 2 HCO3– + H+ → Acetate– + 4 H2O –104.6
ent probes have been developed to target the rRNA
Lactate → 1.5 Acetate + 0.5 H
– – +
–56.5 of different taxonomic groups of SRB93. Mussmann
*Data from REF. 151. et al. 94 used a combination of FISH with catalysed
reporter deposition (CARD–FISH) to study the vertical
distribution of SRB in intertidal mud-flat samples.
in nature (less than 1%) can be cultured. Another They found that up to 11% of all cells were SRB and
classical technique used to determine the presence of that organisms related to the genera Desulfosarcina
SRB in natural samples is the analysis of phospholipid and Desulfobulbaceae dominated the surface layer of
fatty acids 72. This technique has been used to detect the sediment.
groups of SRB, but the taxonomic resolution that can In combination with the use of radioactively labelled
be obtained is limited. Most of the information on substrates, the activity of specific populations can be visu-
the diversity of SRB in both natural and engineered alized. Ito and co-workers95 used a microautoradiography
ecosystems has therefore been obtained by the use –FISH (MAR–FISH) approach to determine the
of marker genes. The most commonly used marker relative abundance of SRB in sewer biofilms and their
Phospholipid fatty acid
A key component of the gene is the gene that encodes 16S ribosomal RNA substrate-uptake patterns in the presence of different
cellular membrane of living (rRNA). electron acceptors. They found that Desulfobulbus was
cells that can be used to Based on comparative analysis of 16S rRNA the most dominant SRB genus in the biofilms, prefer-
identify specific groups of sequences, the known SRB can be grouped into seven entially taking up 14C-propionate and 3H-acetate with
microorganisms and to monitor
their physiological state.
phylogenetic lineages, five within the Bacteria and two sulphate as an electron acceptor, whereas Desulfovibrio
within the Archaea (FIG. 4). Most of the sulphate reducers spp. showed a positive uptake of 14C-bicarbonate in the
CARD-FISH belong to the ~23 genera within the Deltaproteobacteria, presence of hydrogen and sulphate.
Fluorescence in situ followed by the Gram-positive SRB within the Instead of radioisotopes, stable isotope probing (SIP)
hybridization with horseradish
Clostridia (Desulfotomaculum, Desulfosporosinus and can be used to determine the compositions of active
peroxidase-labelled
oligonucleotide probes and Desulfosporomusa genera). Three lineages, Nitrospirae populations. By phospholipid fatty acid analysis of
fluorochrome-labelled (Thermodesulfovibrio genus), Thermodesulfobacteria samples from estuarine sediments that were incubated
tyramides. The tyramides are (Thermodesulfobacterium genus) and Thermodesulfo with 13C-acetate, Boschker et al.96 found that this sub-
deposited at the hybridization biaceae (Thermodesulfobium genus)73, only contain strate was mainly consumed by relatives of the Gram-
site, resulting in enhanced
fluorescence intensity.
thermophilic sulphate reducers. Within the Archaea, SRB positive Desulfotomaculum acetoxidans and not by the
belong to the genus Archaeoglobus in the Euryarchaeota, Gram-negative Desulfobacter spp., as was expected.
Microautoradiography and to the genera Thermocladium74 and Caldirvirga75 in Webster and co-workers97 compared SIP of DNA and
A photographic technique to the Crenarchaeota. phospholipid fatty acids to identify the active com-
visualize the uptake of
Different primer sets have been described76 for the munity constituents in sulphate-reducing sediment
radioactive substrates by
single cells. specific amplification of 16S rRNA gene fragments from enrichments. After short incubations with differ-
different groups of SRB, such as the Desulfotomaculum, ent 13C‑labelled substrates, they found that bacteria
Stable isotope probing Desulfobulbus, Desulfobacterium, Desulfobacter, related to the acetate-utilizing genus Desulfobacter, as
A technique to identify Desulfonema–Desulfosarcina–Desulfococcus and well as a member of the new candidate division JS1,
microorganisms in
environmental samples that
Desulfovibrio genera. A more powerful approach for the which comprises only uncultured members, had taken
have taken up a stable isotope- detection of SRB is the use of so-called functional genes up 13C-acetate. Unfortunately, this result could not be
labelled substrate. which encode enzymes that play an important part in substantiated by phospholipid fatty acid analysis.

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12 Desulfobulbaceae
32 Desulfobacteraceae
Desulfobacterium anilini (AJ237601)

Desulfarculus baarsii (AF418174)
8 Syntrophobacteraceae
Desulfomonile limimaris (AF282177)
Desulfomonile tiedjei (AM086646)
Deltaproteobacteria
47 Desulfovibrio spp.
Desulfonatronum lacustre (AF418171)

Desulfonatronum thiodismutans (AF373920)
Desulfonatronum cooperativum (AY725424)
Desulfonatronovibrio hydrogenovorans (X99234)
Bacteria
7 Desulfomicrobium spp.
Desulfocaldus terraneus (AY464939)
6 Desulfohalobium spp.
0.10 Thermodesulfovibrio islandicus (X96726) Nitrospirae

Thermodesulfovibrio yellowstonii (AB231858)
Desulfosporomusa polytropa (AJ006605)
Desulfosporosinus orientis (AJ493052)
Desulfosporosinus youngii (DQ117470)
Desulfosporosinus meridiei (AF076247) Clostridia
18 Desulfotomaculum spp.
Thermodesulfobium narugense (AB077817) Thermodesulfobiaceae
Thermodesulfobacterium hveragerdense (X96725)
Thermodesulfobacterium thermophilum (AF334601)
Thermodesulfobacterium commune (AF418169) Thermodesulfobacteria
Thermodesulfobacterium hydrogeniphilum (AF332514)
Thermodesulfatator indicus (AF393376)
Archaeoglobus profundus (AF297529)
Archaeoglobus veneficus (AF418181) Euryarchaeota
Archaea
Archaeoglobus lithotrophicus (AJ299218)
Thermocladium modestius (AB005296)
Crenarchaeota
Caldivirga maquilingensis (AB013926)
Figure 4 | Phylogenetic tree based on nearly complete 16S ribosomal RNA (rRNA) sequences of described
sulphate-reducing bacterial species. The sequences were obtained from the SILVA small subunit (SSU) rRNA database
Nature
(version 03 08 22)149 and the tree was created using ARB software150 (see Further information). Note the Reviews | Microbiology
seven phylogenetic
lineages of sulphate-reducing bacteria, two in the Archaea and five in the Bacteria. The number within the collapsed clusters
indicates the number of different species within a particular group. The scale bar indicates 10% sequence difference.
Other tools to study the activity of SRB are the use Distribution of SRB
of microelectrodes for sulphide measurements98 and SRB are not only versatile in their use of various
the use of radiolabelled sulphate to determine sulphate- electron acceptors and electron donors, they can also
reduction rates. Recently, gene-expression studies — for thrive in a range of different environmental condi-
example, the detection of mRNA of genes that encode tions. They are ubiquitous and can be found in many
key enzymes in the sulphate‑reduction pathway — were natural and engineered environments where sulphate
carried out to infer the activity of SRB in natural samples. is present. SRB have been detected or isolated from
Wawer and co-workers99 studied the expression of the marine sediments 94,96,97,100, hydrothermal vents 101,
NiFe hydrogenase gene to infer the niche differentiation of hydrocarbon seeps 26,102 and mud volcanoes 103, and
Niche differentiation
The tendency for coexisting coexisting Desulfovibrio spp., and Dar et al.81 studied the are abundantly present in hypersaline microbial
species to differ in their use of expression of dsrB genes to infer the activity of all SRB. mats, even at saturating oxygen concentrations83,104.
resources. However, all these methods have their advantages and They have been detected in habitats with extreme
disadvantages, and so to obtain a comprehensive under- pH values, such as acid-mine drainage sites, where the pH
Acid-mine drainage site
Acid water that contains H2SO4
standing of the diversity and activity of SRB in their can be as low as 2 (Ref. 105) and in soda lakes, where
derived from microbial natural habitat an integrated approach using different the pH can be as high as 10 (Ref. 82). SRB have been
oxidation of sulphidic minerals. traditional and molecular methods should be used81. detected and isolated from oil fields106, as well as from

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Box 2 | Sulphate reducers and corrosion

Microbially induced corrosion or biocorrosion of
3Fe(OH)2
steel results in huge financial losses that amount to 6 OH+
US$100 million per annum in the United States Anode
alone144. Both aerobic and anaerobic corrosion
FeS 3 Fe2+
occurs. Steel and iron surfaces act as a substratum e– e–
for microbial communities to form biofilms. Owing Fe2+ e– e–
to oxygen consumption by aerobic 4 H2O e–
e– e– e–
microorganisms, biofilms are largely anaerobic at 2 lactate + SO4 2–
S 2–
the metal surface, which creates a niche for H2

anaerobic bacteria. Fermentation of decaying H2 e– e– e– e–
biomass also takes place. Fermentation products, 2 acetate + 2 CO2 + 2 H2O
such as lactate, are used as electron donors for H2 e– e– e– e–
sulphate-reducing bacteria (SRB). At the steel and H+ Cathode
H2
iron surface, electrochemical corrosion occurs (see H+ OH– SO42–
H+
the figure). Chemical dissolution of iron then
results in the formation of hydrogen according to OH–
H+ H+
Equation 9. H+
H+
Fe + 2H+ → Fe2+ + H (9)
SRB consume hydrogen and influence the equilibrium of the chemical dissolution. Iron sulphide is formed as a
Nature Reviews | Microbiology
product of chemical dissolution and sulphate reduction. Some sulphate reducers have the ability to enhance
hydrogen formation from steel (F. Widdel, personal communication). Iron and steel corrosion is most severe in marine
water that contains a high concentration of sulphate. However, SRB also have an important role in freshwater owing
to an active sulphur cycle. It is therefore important that sulphate reduction can occur at freshwater sulphate
concentrations145 because Desulfovibrio species have a high affinity for sulphate71,146. Figure modified, with
permission, from REF. 147  (2005) Taylor & Francis.
the deep sub-surface107. They are also present in fresh- from groundwater and waste water. This application
water sediments108, in the rhizosphere of plants109,110, in takes advantage of differences in the chemical prop-
aquifers and in engineered systems, such as anaerobic erties of metal sulphates and sulphides 116,117. Metal
waste-water treatment plants 69,80,81,90,98,99. Most SRB sulphates (cadmium, cobalt, copper, iron, nickel and
are free-living, but some are present in consortia zinc) are highly soluble, but the corresponding metal
with other microorganisms, such as methanotrophic sulphides have low solubility. Thus, by sulphate reduc-
archaea 29 , or even in a more intimate relation- tion, metals can be precipitated, recovered and reused.
ship, for example, together with sulphur‑oxidizing This concept has been applied to immobilize metals
Gammaproteobacteria as endosymbionts in the marine from surface water and process water from mining
worm Olavius algarvensis 111, thereby providing the industries. Organic waste materials (for example,
host with nutrients112. straw) are often used to immobilize heavy metals in
lake sediments118,119. Defined substrates, such as lactate,
Biotechnological applications of SRB ethanol, methanol and hydrogen-rich gas, are often
Sulphuric acid is used in many industrial processes, preferred as electron donors for sulphate reduction.
which results in the occurrence of sulphate in waste Based on the THIOPAQ system (Paques; see Further
water. Sulphate reduction will therefore occur, which information), a process for sulphate reduction and
is highly undesirable. For example, in anaerobic oxidation of the excess sulphide was developed to remove
treatment of agro–industrial waste waters, sulphate heavy metals (FIG. 5) . The sequence of conversions
reduction results in lower methane yields. In addition, is provided in Equations 1–3.
sulphide is toxic, odorous and corrosive. Attempts to
avoid sulphate reduction by changing the flow regime SO42– + 8 [H] + H+ → HS– + 4 H2O (1)
in methanogenic bioreactors were not successful. Also, in
the petro–chemical industry, sulphate reduction HS– + Me2+ → MeS↓ + OH– (2)
causes problems: hydrogen sulphide formation causes
corrosion (BOX 2) and safety problems for personnel HS– + 0.5 O2 → S° ↓ + OH– (3)
who are involved in offshore activities113. To avoid
hydrogen sulphide formation in oil fields, it has been This process is in operation at a zinc smelter (Nyrstar,
proposed that nitrate should be injected to stimulate The Netherlands) to treat the zinc sulphate‑containing
nitrate-reducing activity to oxidize hydrogen sulphide process water. Sulphate reduction takes place in a
and suppress sulphate reduction by nitrite or by the full-scale (500 m3) sulphate-reducing gas-lift reactor.
higher redox potential that is created114,115. Synthesis gas, which is formed by steam-reforming
Sulphate reduction can be applied beneficially to natural gas, is the electron donor for sulphate reduc-
biotechnology, such as the removal of heavy metals tion. The gas that enters the reactor is composed of 76%

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CO2 In the first step, sulphur dioxide is scrubbed from

the flue gas with an alkaline solution to form sulphite
(Equation 4).
SO42– + 4 H2 + H+ HS– + 4 H2O SO2 + OH– → HSO3– (4)

Zn + HS
2+ –
ZnS + H+
Gas lift The presence of oxygen in the flue gas results in
Bioreactor one Bioreactor two the unavoidable oxidation of part of the sulphite to
Excess sulphate (Equation 5).
Waste water or process HS– Purified
water (SO42– and metals, effluent
such as Zn2+) HSO3– + 0.5 O2 → SO42– + H+ (5)
ZnS So In an anaerobic bioreactor, sulphite and sulphate

Air are reduced by SRB to sulphide (Equations 6,7).
2 HS– + O2 + 2 H+ 2 So + 2 H2O HSO3– + 6 [H] → HS– + 3H2O (6)

H2-rich gas
SO42– + 8 [H] → HS– + 3H2O + OH– (7)
Figure 5 | Schematic overview of the THIOPAQ process to remove sulphate
and heavy metals from waste water. Sulphate is reduced to sulphide by Then, in a micro-aerobic reactor, sulphide is par-
sulphate‑reducing bacteria in bioreactor one using hydrogen as an electron donor. tially oxidized to elemental sulphur by autotrophic
Subsequently, the sulphide is used to precipitate the heavy metals. An excess of sulphide-oxidizing bacteria (Equation 8).
sulphide is converted to elemental sulphur by sulphide-oxidizing bacteria in
bioreactor two. The precipitated metal sulphides and elemental sulphur can HS– + 0.5 O2 → S° ↓ + OH– (8)
be reused.
During sulphide oxidation, alkalinity is produced,
which can be led back to entrap sulphur dioxide.
hydrogen, 20% carbon dioxide, 3% nitrogen and 1% Alkalinity is only lost via the bleed stream. Therefore,
carbon monoxide. The zinc sulphide that precipitates Bio-FGD requires a low input of lime or limestone. At
with the sulphate-reducing biomass is collected in a the laboratory scale, the feasibility of Bio-FGD has been
settler and reused in the roasting process. More than demonstrated125,126. In China, a full-scale Bio-FGD has
95% of the dry weight of the sludge in the bioreactor is recently been initiated by Paques (see Further infor-
zinc sulphide. mation). This Bio-FGD is operated using a citric acid
The microbial community that is present mainly waste stream as electron donor for sulphate and sulphite
consists of heterotrophic SRB that belong to the genera reduction. Citrate is not a common electron donor for
Desulfovibrio and Desulfomicrobium, but low numbers SRB. Furthermore, initial experiments in our laborato-
of methanogens and homoacetogens are also present120. ries indicate that citrate is not the direct electron donor
The dominant SRB require acetate as a carbon source. for SRB: it is first fermented to acetate and formate by
When acetate (or another organic carbon source) is Trichococcus and Veillonella species. Thus, acetate and
not supplied, as in the full-scale reactor, these sul- formate are the direct electron donors for sulphate reduc-
phate reducers will depend on acetate formation by tion. The preferential use of acetate in sulphate reduction
homoacetogens. One of the dominant sulphate reduc- requires long-term operation of the bioreactors70. Several
ers, Desulfovibrio paquesii, which is also abundantly sludge samples from different sulphate-reducing biore-
present in other sulphate-reducing bioreactors121, was actors that are in operation contain high numbers of
isolated and classified as a novel species122. acetate-degrading SRB81.
Another biotechnological application for SRB is
the removal and reuse of sulphur compounds from Concluding remarks and future perspectives
waste water and off gases. Under oxygen-limitation, In this Review, we have discussed how SRB are ubiq-
sulphide-oxidizing bacteria mainly produce elemental uitous in anoxic habitats, where they use sulphate as a
sulphur instead of sulphate (Equation 3). This feature is terminal electron acceptor in the degradation of organic
applied to remove hydrogen sulphide from natural gas compounds. However, their energy metabolism is not
or biogas123,124. When combined with an anaerobic step, restricted to sulphate reduction and SRB can use a wide
it is possible to treat water and gas that contain oxidized range of other electron acceptors. In addition, they also
sulphur compounds. The oxidized sulphur compounds have fermentative metabolism. An important physio
are reduced to sulphide, which is then partially oxidized logical feature of SRB is that they can compete with
to elemental sulphur. One example of this application methanogens or grow syntrophically with them depend-
is flue-gas desulphurization (FGD). Lime or limestone ing on the availability of sulphate. These microorganisms
wet scrubbing is a commonly applied FGD technol- are not only versatile in their metabolism, but also in the
ogy. Biotechnological FGD (Bio-FGD) is an alterna- environmental conditions in which they thrive. The use
tive that makes use of the following conversions of the of different molecular methods has demonstrated that
sulphur cycle. their diversity is enormous and that there are still many

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Table 2 | General genomic features of different sulphate-reducing microorganisms*

Domain Phylum Class Genome G+C Number Predicted Genes Genes CDS Number Number
size (bp) (%) of genes CDSs with without (%) of 16S of
function function rRNAs tRNAs
prediction prediction
Archaeoglobus fulgidus DSM 4304
Archaea Euryarchaeota Archaeoglobi 2,178,400 49 2,519 2,468 1,798 670 98 1 46
Caldivirga maquilingensis IC‑167

Archaea Crenarchaeota Thermoprotei 2,077,575 34 1,986 1,943 1,307 63 98 0 42
Desulfotomaculum reducens MI‑1
Bacteria Firmicutes Clostridia 3,608,104 42 3,424 3,324 2,334 990 97 8 71
Desulfovibrio vulgaris subsp. vulgaris strain Hildenborough

Bacteria Proteobacteria Deltaproteo 3,773,159 63 3,640 3,545 2,216 1,329 97.4 5 68
bacteria
Desulfovibrio vulgaris subsp. vulgaris DP4
Bacteria Proteobacteria Deltaproteo 3,661,391 63 3,189 3,103 2,297 806 97.3 5 68
bacteria
Desulfovibrio desulfuricans G20
bacteria
Desulfotalea psychrophila LSv54
Bacteria Proteobacteria Deltaproteo 3,659,634 47 3,331 3,234 1,739 1,495 97 7 64
bacteria
Synthrophobacter fumaroxidans MPOB
bacteria
*Data from the Integrated Microbial Genomes (IMG) database (see Further information). CDS, coding sequences; rRNA, ribosomal RNA.
uncultured representatives. Apart from their importance One of the greatest challenges in microbial ecology
in nature, SRB, together with sulphur-oxidizing micro is to identify the function of microorganisms in their
organisms, can be successfully exploited in the sustainable natural habitats. MAR–FISH128 and SIP129 have been
clean-up of industrial waste streams. used successfully for this purpose. However, Li et al.130
Although we have generated a huge amount of infor- recently described a new approach, SIMSISH (second-
mation on the diversity, physiology and biochemistry of ary ion mass spectrometry in situ hybridization), that
SRB, we think that we have only scratched the surface. combines probe-based hybridization with isotope
So far, diversity studies have been mainly descriptive, measurements at the single-cell level using a NanoSIMS
and the physiology and biochemistry of SRB have been instrument. If this approach can also be combined with
studied primarily with just a few model organisms, such the in situ detection of mRNA131, it will soon be possible
as Desulfovibrio desulfuricans, Desulfovibrio vulgaris and to study the ecophysiology of SRB, particularly those
Desulfovibrio gigas. Therefore, future research should that have not yet been isolated, in greater detail.
move away from descriptive studies and focus on expla- The complete genomes of different SRB —
nations and predictions, using ecological concepts and Archaeoglobus fulgidus VC‑16132, Caldivirga maquilin-
innovative technologies, such as meta-transcriptomics gensis IC‑167, Desulfovibrio vulgaris subsp. vulgaris
and meta-proteomics. strain Hildenborough133, Desulfotalea psychrophila134,
Although difficult, isolation of microorganisms is still Desulfovibrio desulfuricans G20, Desulfotomaculum
necessary to obtain detailed insights into their physiol- reducens MI‑1 and Syntrophobacter fumaroxidans MPOB
ogy, behaviour and interactions with other organisms, — have been sequenced (TABLE 2), and the genomes of
as well as for biotechnological purposes. Novel high- other SRB, for example, Desulfobacterium autotrophi-
throughput technologies might increase the success of cum, Desulfovibrio magneticus, Thermodesulfovibrio
isolating ecologically important community members. yellowstonii and Thermodesulfobacterium commune, are
Recently, Ingham and co-workers127 described the devel- currently being sequenced. Comparative analysis of these
opment of the micro-Petri disk, a million-well dispos- genomes will provide detailed information on the energy
able chip for culturing and high-throughput screening and carbon metabolism of these organisms, and on the
microorganisms. The use of this revolutionary tool diversity and evolution of the enzymes that are involved
might result in the isolation of different novel SRB. in these processes (BOX 3). Moreover, these sequences

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Box 3 | Genomics of sulphate reducers

Archaeoglobus fulgidus DSM 4304 dsrA B
Caldivirga maquilingensis IC-167
Desulfotomaculum reducens MI-1
Desulfovibrio vulgaris subsp. vulgaris strain Hildenborough
Desulfovibrio vulgaris subsp. vulgaris DP4
Desulfovibrio desulfuricans G20
Desulfotalea psychrophila LSv54
Synthrophobacter fumaroxidans MPOB
The complete genome sequences of eight sulphate reducers have been deposited in publicNature Reviews
databases | Microbiology
to date —
Archaeoglobus fulgidus DSM 4304 (Euryarchaeota), Caldivirga maquilingensis IC-167 (Crenarchaeota), the Gram-
positive bacterium Desulfotomaculum reducens MI-1 (Firmicutes) and five Gram-negative Deltaproteobacteria,
Desulfovibrio vulgaris subsp. vulgaris strain Hildenborough, Desulfovibrio vulgaris subsp. vulgaris DP4, Desulfovibrio
desulfuricans G20, Desulfotalea psychrophila LSv54 and Syntrophobacter fumaroxidans MPOB. The genomes of these
sulphate-reducers have different features (TABLE 2). The genomes of the two archaea, A. fulgidus (~2.2 Mb) and
C. maquilingensis (~2.1 Mb), are much smaller than those of the sulphate-reducing bacteria (~3.6–4.9 Mb) and have a
lower number of transfer RNAs. Comparative analysis of the clusters of orthologous group (COG) profiles (see the
figure) shows a low correlation value of 0.30 or less between the sulphate-reducing archaea and the sulphate-reducing
bacteria. Intermediate values (0.54–0.74) were found among the six bacteria, whereas high correlation values (0.91– 0.99)
were found among the three Desulfovibrio strains. The low similarity between the genomes of A. fulgidus and
D. psychrophila was also observed by Rabus et al.134, who found that only genes that encode proteins which are involved
in sulphate reduction and some common hypothetical proteins were shared, which indicated that only a small fraction
of genes are necessary for sulphate reduction. Comparative analysis of the genomes of other sulphate reducers and
closely related microorganisms is needed to confirm this assumption.
The figure shows the gene neighbourhood of dsrAB genes in different sulphate-reducing bacteria. Genes in the same
colour (except for pale yellow) are from the same COG group.
open up the possibility for functional genomics. DNA allowing their performance in the removal of sulphur
microarrays have been used to study the expression of compounds from waste streams to be improved. SRB
genes under different environmental conditions, such as have been studied successfully for more than a century,
temperature, salinity135 and pH, and proteomics has been but the novel opportunities that have been created by
used to study the oxygen stress response136. With these the genomics revolution will generate enormous oppor-
tools in hand, we can not only obtain important informa- tunities for microbiologists to obtain detailed insights
tion on the niche differentiation of SRB, but also predict into the ecology and biotechnology of these important
their behaviour in engineered ecosystems, thereby microorganisms.

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REVIEWS

The ecology and biotechnology of

nature reviews | microbiology volume 6 | june 2008 | 441

Dry deposition of Acidic precipitation

Gypsum and pyrite in Microorganisms

442 | june 2008 | volume 6 www.nature.com/reviews/micro

As many SRB are able to grow on hydrogen and

lactate is first converted to acetate, carbon dioxide and

hydrogen; the hydrogen diffuses out of the cell

and is used as an electron donor for sulphate reduction.

This model, even today, is controversial, but has never

been refuted or confirmed convincingly. Hydrogen

formation during growth on lactate might reflect the

high investment of ATP to transport sulphate across

nature reviews | microbiology volume 6 | june 2008 | 443

444 | june 2008 | volume 6 www.nature.com/reviews/micro

Box 1 | Sulphate reducers and oxygen

nature reviews | microbiology volume 6 | june 2008 | 445

446 | june 2008 | volume 6 www.nature.com/reviews/micro

Desulfobacterium anilini (AJ237601)

Desulfonatronum lacustre (AF418171)

0.10 Thermodesulfovibrio islandicus (X96726) Nitrospirae

nature reviews | microbiology volume 6 | june 2008 | 447

Box 2 | Sulphate reducers and corrosion

the metal surface, which creates a niche for H2

448 | june 2008 | volume 6 www.nature.com/reviews/micro

CO2 In the first step, sulphur dioxide is scrubbed from

SO42– + 4 H2 + H+ HS– + 4 H2O SO2 + OH– → HSO3– (4)

ZnS So In an anaerobic bioreactor, sulphite and sulphate

2 HS– + O2 + 2 H+ 2 So + 2 H2O HSO3– + 6 [H] → HS– + 3H2O (6)

nature reviews | microbiology volume 6 | june 2008 | 449

Table 2 | General genomic features of different sulphate-reducing microorganisms*

Caldivirga maquilingensis IC‑167

Desulfovibrio vulgaris subsp. vulgaris strain Hildenborough

450 | june 2008 | volume 6 www.nature.com/reviews/micro

Box 3 | Genomics of sulphate reducers

Caldivirga maquilingensis IC-167

Desulfotomaculum reducens MI-1

Desulfovibrio vulgaris subsp. vulgaris strain Hildenborough

Desulfovibrio vulgaris subsp. vulgaris DP4

Desulfovibrio desulfuricans G20

Desulfotalea psychrophila LSv54

Synthrophobacter fumaroxidans MPOB

nature reviews | microbiology volume 6 | june 2008 | 451

452 | june 2008 | volume 6 www.nature.com/reviews/micro

nature reviews | microbiology volume 6 | june 2008 | 453

454 | june 2008 | volume 6 www.nature.com/reviews/micro

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