Journal of Evolutionary Biochemistry and Physiology, Vol. 37, No. 4, 2001, pp. 444—450.

Translated from Zhurnal Evolyutsionnoi Biokhimii i Fiziologii,

Vol. 37, No. 4, 2001, pp. 338—343.
Original Russian Text Copyright © 2001 by Verkhovtseva, Filina, Pukhov.

PROBLEM PAPERS

Evolutionary Role of Iron in Metabolism of Prokaryotes

and in Biogeochemical Processes1
Yu. V. Verkhovtseva*, Ya. Yu. Filina**, and D. E. Pukhov**
* Lomonosov State University, Moscow, Russia
** Demidov State University, Yaroslavl, Russia
Received April 12, 2000

Abstract—The paper reviews data summarizing points of view about the “conceptual” role of iron
in the appearance and evolutionary formation of the Earth and its biosphere. Partici pation of iron
and its compounds in the appearance and development of processes of anaero- and aerobiosis as
fundamental “blocks” of metabolism is presented as a hierarchical scheme. Magnetically arrayed
iron compounds, in which the element is both in the Fe(II) and in the Fe(III) state, are considered
a connecting link between the hierarchical levels. It is shown that the energy transformation Fe(II) ↔
Fe(III) is an oxidation–reduction energy core of the most important metabolic iron complexes and
of processes of biogenesis both at the cellular level and in biogeosystems.

IMPORTANCE OF IRON IN FORMATION The subsequent accumulation of oxygen in at-

OF BIOSPHERE ON THE EARTH
According to modern concepts of some geolo-
gists, geochemists, biochemists, and microbiol-
ogists, the whole evolution of the Earth and pos-
sibility of origin of life on it depended essentially
on the ratio of Fe and FeO in the Earth crust [1–
4], i.e., on the ratio of amounts of metallic iron
and Fe(II), which in the primordial Earth mat-
ter amounted to 13 and 24%, respectively. In
the early Earth history these iron forms were the
main absorbents of oxygen formed at photolysis
of water, the metallic iron amount decreasing in
the process of its transformation into the Fe(II)
state. In the arising anoxygenic prokaryotic life
(about 4 bln years ago) the iron was used in the
form of Fe(II) compounds (for example, FeS-
proteins and ferredoxin (EC 1.8.7.1)) as electron
carriers in strict heterotrophic anaerobic fermen-
tators [5].
mosphere (due to oxygenic photosynthesis by cy-
anobacteria) was accompanied by its binding by
Fe(II) iron oxide and conversion to Fe(III).
With the accumulation of free oxygen in the Earth
atmosphere a shift to the next oxygenic stage of
life evolution, iron was included in metabolic
pathways both in the Fe(II), and in Fe(III)
state. The importance of such compounds is due
to ability of their “iron–oxide” core to accept
or release electrons in the course of transforma-
tion Fe(II) ↔ Fe(III), which, thereby, is the
energy center. Partici pation in diverse oxidation–
reduction reactions determines the essential im-
portance of iron complexes both at the level of
metabolic transformations in the cell, and in bio-
geochemical processes. These levels are consid-
ered below in a greater detail.
1 The paper is published as a matter of discussion.

0022-0930/01/3704-0444$25.00 © 2001 MAIK “Nauka/Interperiodica”

 EVOLUTIONARY ROLE OF IRON IN METABOLISM OF PROKARYOTES
BIOCHEMICAL PROCESSES OF IRON
TRANSFORMATIONS AT THE CELLULAR
LEVEL
The iron occupies a special place among 12
macroelements necessary for the life activity of
both prokaryotic, and eukaryotic organisms at a
relatively high (> 10–4 M) concentration. The
significant role of iron in metabolism is due to its
peculiarity as a transient element that can easily
change its oxidation degree, Fe(II) or Fe(III).
It allows iron to coordinate various electron do-
nors (coordination ligands) and to partici pate in
various oxidation–reduction processes. Ligands
form various surrounding of the core, which de-
termines formation of iron complexes, particu-
larly, of iron-containing proteins, in nature.
Owing to the wide occurrence, as well as to their
properties, these compounds were included in
fundamental processes of metabolism in organ-
isms: in DNA synthesis (ribonucleoside di phos-
phate reductase (EC 1.17.4.1) and ribonucleoside
tri phosphate reductase (EC 1.17.4.2)), in photo-
synthesis, respiration, nitrogen fixation, and some
others [6]. At present, more than 20 of such
metabolically important biocomplexes have been
known. As compared with other metals (Mg,
Ca, Co, Cu, Zn, Mo) forming metallobiomol-
ecules in organisms, only iron forms such diver-
sity of biocomplexes [7].
In the scheme I we have tried to present the
main stages of evolution of biosphere, anaerobic
and aerobic, and to specify fundamental process-
es of life activity of prokaryotes and the protein
iron complexes that determine the occurrence of
these processes.
Considering evolutionary development of
prokaryotes, bacteria and cyanobacteria, that
prevail on the Earth during more than 3 bln years
and are “test objects of the Nature” in its search
for optimal biochemical pathways of metabolism,
a direct partici pation of iron compounds can be
noted in this natural selection of metabolism. In-
deed, protein complexes of iron partici pate in all
fundamental processes of the life activity both at
anaerobic, and at aerobic stages of evolution of
biosphere (scheme I).
Thus, such “ancient” protein complexes of
iron, as FeS-proteins, ferredoxin, rubredoxin (EC
JOURNAL OF EVOLUTIONARY BIOCHEMISTRY AND PHYSIOLOGY
445
1.18.1.1), hydrogenase (EC 1.18.99.1), cyto-
chrome c oxidase (EC 1.9.3.1), are involved in
the most important energy processes of anaerobi-
osis, fermentation evolutionary more recent pho-
tosynthesis in eubacteria. The same enzymes pro-
vide anaerobic nitrogen fixation. The iron-con-
taining enzymes are also necessary in anaerobic
respiration: “nitrate” and “sulfate”: nitrate reduc-
tase (EC 1.7.99.4) and nitrite reductase (EC
1.7.99.3), and hydrogenase, respectively. Besides,
there are processes of biogenesis of iron compounds
at the anaerobic stage of life, i.e., extra- and in-
tracellular formation of iron minerals with par-
tici pation of bacteria. More than 10 such biom-
inerals have been described [8]. Of a special in-
terest is the ferrimagnetic mineral magnetite
(FeO2 • Fe2O3) that is composed both of Fe(II),
and of Fe(III), i.e., at the biochemical level the
gradient (microaerobic) conditions occur, under
which the energy balance in relation to the iron
mineral formation is determined by the ratio
Fe(II) : Fe(III). The problem of the biomineral-
ization process and the magnetite biogenesis is con-
sidered below in a greater detail.
In the oxygen atmosphere, spectrum of iron
complexes partici pating in metabolism of prokary-
otes was extended, while Fe-enzymes of anaero-
bic stage of the biosphere development were pre-
served. By their example it is possible to follow
“the evolution itself that looks like a gradual com-
plication, whose previous stage is included in the
subsequent one as a necessary constituent” [9].
Thus, the aerobic transport chain was composed
of iron-containing dehydrogenases: succinate de-
hydrogenase (EC 1.2.1.16), NADH dehydroge-
nase (EC 1.6.99.2), formiate dehydrogenase (EC
1.2.1.43), and cytochromes. The defense from
reaction oxygen compounds was required, so such
iron complexes as superoxide dismutase (EC
1.15.1.1), xanthine oxidase (EC 1.2.3.2), cata-
lase (EC 1.11.1.6), peroxidase (EC 1.11.1.7) were
“selected” in the process of evolution to perform
this task. Cofactor of the above-listed enzymes is
iron with the (III) oxidation state, whose revers-
ible reduction to Fe(II) determines their func-
tioning. Besides, catalase and peroxidase contain
iron in the heme structure.
The oxygen stage of the nitrogen cycle, apart
from Fe-enzymes partici pating in anaerobic ni-
Vol. 37 No. 4 2001
446 VERKHOVTSEVA et al.

JOURNAL OF EVOLUTIONARY BIOCHEMISTRY AND PHYSIOLOGY Vol. 37 No. 4 2001

 EVOLUTIONARY ROLE OF IRON IN METABOLISM OF PROKARYOTES
trogen fixation, includes leghemoglobin (EC
1.6.2.6), providing oxygen binding and defense of
the nitrogenase complex of symbiotic nitrogen fix-
ers from its reaction action. As a whole, at this
period there is an extension of spectrum of iron-
containing enzymes with the heme structure that
has begun to be the main structure for Fe-com-
plexes in eukaryotes in the process of evolution.
Thus, in the evolutionary hierarchy of iron-
containing compounds, which we have proposed
(scheme I), in fundamental “blocks” of metabo-
lism, it is possible to trace a transition from Fe(II)-
compounds in anaerobic processes to Fe(III)-com-
plexes in aerobic ones, whose fine energy balance
is associated with the Fe(II) : Fe(III) ratio and
determines the cell biochemical state. We con-
sider the hierarchy of iron compounds as a rise
of their rank owing to internal rearrangements,
particularly due to involvement of other ligands
in the complexes at change of the degree of the
metal oxidation.
When considering the iron cycle at the bio-
geochemical level, there seems reasonable the
concept of importance of the ratio of the iron
concentration and the (II) and (III) oxidation
states as an ecological factor that determines buffer
capability in microzones at the phase boundary,
under the so-called gradient conditions [10, 11]
forming geochemical barriers. These barriers are
peculiar “sites” of the earth crust, which have
been used by Nature to model geochemical con-
ditions (“situations”) with specific physical-chem-
ical characteristics that provide a local concen-
tration of some compounds: these are ecological
habitation areas that are the most favorable for
biogenesis.
BIOGEOCHEMICAL CYCLE OF IRON
Iron occupies the second place among metals
and the fourth among elements by the per cent
contents (4.65%) in the earth crust. Iron is an
important bioelement, its per cent contents in
the living matter amounts to 1 × 10–2%.
The richest in natural iron deposits are the pre-
Cambrian oxidized sediments, known as blended
irons formations (BIFs) that concentrate about
28% of the earth crust iron. They represent alter-
nating layers of siliceous rock containing magne-
JOURNAL OF EVOLUTIONARY BIOCHEMISTRY AND PHYSIOLOGY
447
tite and goethite [14]. These formations are of
sea origin, their age is 3.2–1.9 bln years [15].
Other crystalline iron reserves are bog iron ores,
”red stratas”—continental or marginal deposits,
in which fine grains of silicon dioxide are covered
by iron oxides [15], and also hydrothermal de-
posits formed as a result of volcanic and mag-
matic activity, and iron–manganese deposits of
lakes and oceans, but all of them are quantita-
tively less than BIFs.
The biogeochemical iron cycle in nature is a
complex of the global “slow” geochemical cycle
and minor biological turnovers performed with
partici pation of living organisms, mainly micro-
organisms. The main part of iron in the modern
geosphere is in the oxidized state that is stable for
this element under the most predominant aerobic
conditions with the neutral and low-alkaline re-
action. Therefore, the global iron cycle is a slow
geological cycle characterized by disposals, meta-
morphoses, volcanism, weathering, and other
physical-chemical mechanisms of transformation
and transport.
However, there are in the nature such condi-
tions, in which the iron cycle proceeds rather
fast. These are reductive or partly anaerobic
zones, in which reduction of iron and its mobili-
zation is observed. The examples of such zones
are bottoms of eutrophized and stratified lakes,
seasonal anaerobic ponds, partly anaerobic de-
posits of the marine and freshwater origin, moors,
etc.
In performance of stages of the “fast” iron cy-
cle, the determining role is played by microor-
ganisms interacting with iron of geological depos-
its at gradient geochemical barriers. Such micro-
organisms are believed to be the main partici pants
of the BIFs formation [16].
Taking into account biochemical transforma-
tions of iron at the cellular level and biogeochem-
ical processes of global geological cycle, the par-
tici pation of bacteria in the iron turnover may be
presented as follows (Scheme II). A link between
the cellular and biogeochemical levels are Fe(III)-
compounds that predominate in nature and are
low soluble at ðÍ > 3.0. Thus, concentration of
Fe(III) ions of hydroxides in water amounts to
around 10–17 M [19]. Prokaryotes have devel-
oped various mechanisms of the iron transport and
Vol. 37 No. 4 2001
448 VERKHOVTSEVA et al.

Scheme II. Partici pation of bacteria in iron cycle [16–18]

Få(II) –Få(III)-complexes absorbed by cell

B
I P
O R
BIOMINERALIZATION BASAL METABOLISM C
(hydroxides, bacterioferritin, ferrihydrite, Siderophoric binding O
(iron-containing enzymes) H
pyrrhothine, magnetite, gragite) C
E E
M S
I S
Clearing at dying Clearing at dying C E
A S
Fe(III)-compounds: L
oxides, hydroxides etc.
(neutral conditions of medium) Fe(II), Fe(III)
Precipitation B
and sedimentation metabolism
I
O
G P
Fe(III)-minerals, Fe(II) Fe(III) Fe3O4 Få(III) E R
FeOOH, Fe2O3, Fe(OH)3 dissolution deposition O O
Magnetite
C C
Reduction Oxidation production Fe-reduction Fe-oxidation
(neutral H E
Gragite (low pH S
Fe3S4 conditions of medium) E
production M S
of medium)
E
I
C S

FeS Få(II), A
Buried minerals deposition H2SO4 L
S-oxidizing bacteria

Disposal and diagenesis

absorption to provide their metabolic needs in iron
[20]. Many microorganisms are able to release
siderophores into the environment; these form
complexes at interaction with iron ions (sidero-
phoric binding). By such a way the transport of
iron into the bacterial cell is the most favorable.
The Fe-ion absorbed as a result of the specific
transport is used in basic processes of metabolism
and in biogenesis (in some species). After dying,
the cell and the iron-containing complexes formed
by the cell enter the environment and are involved
in the biogeochemical cycle. Besides, microor-
ganisms partici pate indirectly in transformation of
iron compounds due to a release of mucus, for-
mation of capsules and changes of extracellular
ðÍ in the process of metabolism. This results in
a passive deposition of several iron-containing
minerals on the cell surface and in the environ-
ment [21]. At the biogeochemical level, of sig-
nificance are processes of preci pitation, sedimen-
tation, and Fe(II)/Fe(III) metabolism in micro-
bial biocenoses. The closure of the biogeochemi-
cal cycle is provided by the Fe(II) oxidation and
Fe(III) reduction, the disposal and diagenesis

JOURNAL OF EVOLUTIONARY BIOCHEMISTRY AND PHYSIOLOGY Vol. 37 No. 4 2001

 EVOLUTIONARY ROLE OF IRON IN METABOLISM OF PROKARYOTES
being possible). On biogeochemical barriers (gra-
dients of alkaline-acidic and oxidation–reduction
potentials), a partial reduction of Fe(III) with
formation of magnetite is possible. In some cas-
es, in such compounds there is an isomorphous
substitution of oxygen by sulfur with formation of
pyrrhothine (Fe1–xS, where õ varies from 0 up
to 0.2) and gragite (Fe3S4). All these minerals
have strong magnetism [22].
It is to be reminded that when considering bio-
chemical processes of transformation of iron at
the cellular level, we also should note the extra-
and intracellular formation of magnetite, pyrroth-
ine, gragite, and some other minerals (scheme I).
Apparently, there are common mechanisms of
biogenesis of such minerals at the cellular level,
and also as a result of interaction of abiogenic
and biogenic pathways of formation of these min-
erals at the level of biogeochemical transforma-
tions. Comparing energy values (∆G0) of the dy-
namic system of magnetite formation with partic-
i pation of bacteria under anaerobic conditions
(1) and during weathering (2), a much higher
efficiency of the first process can be noticed:
(∆G0), kJ/mol
ÑÍ3ÑÎΖ + 24 Fe(OH)3 =
8 Fe3O4 + ÍÑÎ3– + 37 H2O (–712) [ 23 ], (1)
Fe2O3 + H2O + 2 e– =
2 Fe3O4 + 2 OH– (–442) [24]. (2)
The magnetite is considered in biomineralogy as
a mineral that is spread sufficiently wide in living
organisms [8], including human [25, 26], whereas
under abiogenic conditions it is formed in meta-
morphic and magmatic deep rocks, scarns, as well
as in middle- and high-temperature hydrothermal
deposits, i.e., where there is a source of exogenous
energy. It indicates once again an importance of
mechanisms of biological induction and control of
biogenesis of this mineral, which seem to have de-
veloped in the process of evolution. Besides, it is a
perfect example of a strong interrelation of living
organisms and the inert matter in the biogeosys-
tem, when considering the concept of the iron cy-
cle in nature. The basis of this interrelation is the
energy transformation Fe(II) ↔ Fe(III) that deter-
mines turnover of biochemical energy both at the
cellular and at the biogeosystemic levels and in the
process of evolutionary biochemical adaptation.
JOURNAL OF EVOLUTIONARY BIOCHEMISTRY AND PHYSIOLOGY
449
CONCLUSION
The exposed above allows stating the key role
of iron in the appearance and evolutionary devel-
opment of the Earth and its biosphere. The role
of this element may be expressed as the following
hierarchical chain of historical transformation:
metallic Fe (Earth core and mantle) → metallic
Fe and FeO oxide in primordial regolith and vol-
canic ashes (a catalyst of synthesis of organic com-
pounds) → Fe/Fe(II)-relation, where oxidation
occurs at binding of O2 formed at photolysis of
water → Fe(II)/Fe(III)-relation, where oxida-
tion of Fe(II) in Fe(III) occurs by binding of O2
formed as a result of oxygenic photosynthesis by
cyanobacteria → Fe(II) ↔ Fe(III) energy trans-
formation that is the oxidation–reduction energy
“core” of the most important metabolic iron com-
plexes and processes of biogenesis both at the cel-
lular level and in biogeosystems, which has de-
termined the princi pal role of iron in nature. As
a connecting link between these levels, magneti-
cally arrayed minerals of iron may be considered,
in which the element exists both in the Fe(II)
and in the Fe(III) state.
ACKNOWLEDGMENTS
The work is supported by the program “Univer-
sities of Russia” (project no. 1169).
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