271
Bioremedial potential of microbial mechanisms of metal
mobilization and immobilization
Geoffrey Michael Gadd
Microorganisms play important roles in the environmental fate
of toxic metals and radionuclides with a multiplicity of
mechanisms effecting transformations between soluble and
insoluble forms. These mechanisms are integral components of
natural biogeochemical cycles and are of potential for both in
situ and ex situ bioremedial treatment processes for solid and
liquid wastes.
Addresses
Department of Biological Sciences, University of Dundee, Dundee,
DD1 4HN, UK; e-mail: g.m.gadd@dundee.ac.uk
Current Opinion in Biotechnology 2000, 11:271–279
0958-1669/00/$ — see front matter
© 2000 Elsevier Science Ltd. All rights reserved.
Abbreviations
APCR air pollution control residues
CP Gly–Cys–Gly–Cys–Pro–Cys–Gly–Cys–Gly
DMSe dimethyl selenide
HP Gly–His–His–Pro–His–Gly
PCS phytochelatin synthase
SRB sulfate-reducing bacteria
Introduction
The mechanisms by which microorganisms effect changes
in metal speciation and mobility are fundamental compo-
nents of biogeochemical cycles for metals, as well as all other
elements including carbon, nitrogen, sulfur and phosphorus,
with additional implications for plant productivity and
human health [1••]. Almost all metal–microbe interactions
have been examined in the context of environmental
biotechnology as a means for removal, recovery or detoxifi-
cation of inorganic and organic metal or radionuclide
pollutants [2,3,4••,5••]. Although much research is laborato-
ry based, there have been many developments to
pilot/demonstration scale, with some processes apparently
in successful commercial operation. It should also be noted
that metal removal/transformation processes are intrinsic
though less appreciated components of traditional means of
water/sewage treatment as well as reed bed, lagoon and wet-
lands technologies [4••,6]. Molecular and genetic analysis is
now furthering our understanding of microbial metal metab-
olism, including those aspects that have potential in
biotechnology [7••,8••]. This review details recent advances
in understanding of the most important mechanisms of
microbial metal transformations within the contexts of
bioremediation and environmental biogeochemistry.
Metal mobilization
Leaching
Leaching of mineral ores by acidophilic sulfur-oxidizing bac-
teria is an established bioindustry and although most interest
arises from a hydrometallurgical perspective, leaching of
contaminating metals from soils and other matrices is also
possible [9••]. In fact, bioleaching using elemental sulfur as
substrate can be better than sulfuric acid treatment for metal
solubilization from contaminated aquatic sediments [10].
Iron- and sulfur-oxidizing bacteria, Thiobacillus ferrooxidans
and Thiobacillus thiooxidans, respectively, were enriched
from contaminated soil and were able to leach >50% of the
metals present (As, Cd, Co, Cu, Ni, V, Zn, B, and Be). Strains
of T. ferrooxidans were able to remove all of the Cd, Co, Cu
and Ni [11]. T. ferrooxidans has also been used to treat air pol-
lution control residues (APCR; fly ash and used lime).
Although viability of the Thiobacilli was poor, removal of as
much as 95% of Cd (from APCR of ~270 mg Cd kg–1) was
achieved with ~69% Pb removal (from APCR of
~5 g Pb kg–1) and similar removals for Zn and Cu [12]. Note
that the metals in such leachates can be subsequently
removed using bacterial sulfate reduction where the soluble
metals are precipitated as insoluble sulfides [9••].
The microbial production of organic acids, under the gen-
eral heading of ‘heterotrophic leaching’, is also of
developing interest in pollution treatment and metal recov-
ery, and also in the context of mineral fertilization to
increase plant productivity [1••,13,14]. In the soil, het-
erotrophic metal mobilization is of more importance than
autotrophic leaching and solubilization of, for example,
phosphate- and sulfate-containing minerals, as this leads to
release of these important nutrients [1••]. Heterotrophic
leaching may be particularly appropriate for wastes of high
pH, such as filter dusts/oxides from metal processing as
most Thiobacilli cannot solubilize effectively above pH 5.5.
Fungi need good aeration and a carbon source so their use
is generally envisaged to be in bioreactors. In most fungi,
leaching is mediated by the production of organic acids,
which provide a source of protons and metal-complexing
organic acid anions [1••,13,14]. Trichoderma harzianum can
solubilize MnO2, Fe2O3, metallic zinc and rock (mostly Ca)
phosphate with both chelation and reduction being
involved in oxide solubilization [15]. Effective leaching of
a variety of wastes and low-grade minerals has been
demonstrated, for example, soils and muds, filter
dust/oxides, lateritic ores, copper converter slag, fly ash and
electronic waste materials [1••,16•]. For the latter, a two-
stage process was proposed where initial growth of
solubilizing organisms is followed by separate contact of
electronic scrap with metabolite-containing medium,
which would overcome any inhibition of organism growth
by high scrap concentrations [16•]. Another method for
treatment of metal-contaminated sandy soil relies on
siderophore-mediated metal solubilization by Alcaligenes
eutrophus. Solubilized metals are adsorbed to the biomass
and/or precipitated, with biomass separated from a soil slur-
ry by a flocculation process. This process resulted in a large
decrease in the bioavailability of Cd, Zn and Pb [17••].
272 Environmental biotechnology
Heterotrophic solubilization can also have consequences for
other remedial treatments for contaminated soils.
Pyromorphite, Pb5(PO4)3Cl, is a stable lead mineral and can
form in urban and industrially contaminated soils. Such insol-
ubility reduces lead bioavailability and the formation of
pyromorphite has been suggested as a remediation technique
for lead-contaminated land, if necessary by means of phos-
phate addition. Pyromorphite can be solubilized, however, by
phosphate-solubilizing fungi (e.g. Aspergillus niger) and plants
grown with pyromorphite as a sole phosphorus source accu-
mulate both P and Pb [18••]. Further, during the fungal
transformation of pyromorphite, the biogenic production of
lead oxalate dihydrate was observed for the first time [18••].
This study emphasises the importance of considering micro-
bial processes when developing remediation techniques for
metal-contaminated soils. Mechanisms of lead solubilization
or immobilization such as lead oxalate may have significant
consequences for metal mobility and transfer between envi-
ronmental compartments and organisms.
Related to heterotrophic solubilization is fungal transloca-
tion of, for example, Cs, Zn and Cd, which can lead to
concentration of metals and radionuclides in specific
regions of the mycelium and/or in fruiting bodies. Whether
the concentration factors observed in vitro can be repro-
duced in the field and whether such amounts can
contribute to soil bioremediation remains a topic for further
investigation [19]. Some parallels may be drawn with the
phytoremediatory approach to soil contamination. Other
possibilities relate to the use of copper-tolerant wood decay
fungi to degrade copper-treated wood products, although
results obtained so far have shown only slight effects on the
copper content of wood before or after decay [20].
Alkylation and reduction
Volatile dimethyl selenide (DMSe) can form naturally
from inorganic Se species in soils and this can regulate geo-
chemical cycling of Se and therefore influence Se
bioremediation. Many environmental and soil factors
(e.g. organic amendments and frequent tillage) can be
optimized to increase diffusive transport through soil and
enhance volatilization [21–23]. In moist or flooded soil,
however, less DMSe is lost to the atmosphere, with a sig-
nificant proportion of DMSe being converted to
non-volatile forms of Se. Oxidized dimethylated Se
(ODMSe), including dimethyl selenoxide (DMSeO)
and/or dimethyl selenone (DMSeO2), was the dominant
form of non-volatile Se, and accounted for up to 90% of Se
in flooded soils. Such transformation of DMSe can be
responsible for the low Se volatilization rate in flooded
soils and sediments [24••]. Microorganisms enhance
arsenic mobilization from soil by up to 24%, compared to
formaldehyde-treated controls, with only low formation of
dissolved methylated arsenic species [25]. Other bacteria
are capable of the oxidation of As(III) to As(V) [26].
Fe(III) and Mn(IV) oxides absorb metals strongly and this
may hinder metal extraction from contaminated soils.
Microbial reduction of Fe(III) and Mn(IV) could be one
way for releasing such metals and this process can be
enhanced with the addition of humic materials, or related
compounds. Such compounds may act as electron shuttles
for, for example, U(VI) and Cr(VI), converting them to
less soluble forms, especially if located in tight pore
spaces where microorganisms cannot enter [3]. Bacterial
Fe(III) reduction resulted in release of, for example, Mn
and Co, from goethite, where 5% of the iron was substi-
tuted by these metals [27]. Mercuric reductase, from a
recombinant Escherichia coli strain, has been immobilized
on a chemically modifed diatomaceous earth support, with
immobilization enhancing stability and reusability: maxi-
mal activity was 1.2 nmol Hg mg–1 protein s–1 at an initial
Hg2+ concentration of 50 µmol dm–3 [28••].
Metal immobilization
Biosorption
Biosorption encompasses those physico-chemical mecha-
nisms by which metal species, radionuclides and so on, are
removed from aqueous solution by microbial biomass or
products. The approach can be multidisciplinary as the
basic nature of experimentation is attractive to biologists,
chemists and engineers, although inadequate dialogue
between such groups has lead to considerable superficiali-
ty and duplication in the literature. It has also lead to the
submersion of many other metal accumulation or transfor-
mation processes, including those dependent on
metabolism, under this general subject heading.
Biosorption technology has been extended to almost every
other kind of biological material and even to describe
removal of certain organic substances. Many studies have
demonstrated the efficiency of metal removal by microbial
biomass in a range of reactor formats and under a range of
physical and chemical conditions [29–35]. Recent develop-
ments in reactor formats and/or biomass separation include
hollow-fibre crossflow microfiltration using Pseudomonas
aeruginosa for biosorption of Pb, Cu and Cd in single and
ternary metal systems. Removal efficiencies in both single
and ternary biosorption were Pb>Cu>Cd and at an influent
metal concentration of 200 µM and flow rate of 350 ml h–1,
effluent concentrations of Cu and Pb satisfied discharge
regulations. This study developed a rapid-equilibrium
model and a mass-transfer model that accurately described
single- and multi-metal biosorption, respectively [36•].
Dispersed air flotation was used to separate Cd-laden
actinomycete biomass. At a Cd concentration of 5 mg L–1
and 0.5 g L–1 freeze-dried biomass, Cd removal was virtu-
ally 100% over a wide range of pH values [37]. Arthrobacter
sp. was entrapped within a macroporous and microporous
matrix of (poly)hydroxyethyl methacrylate cross-linked
with trimethylolpropanetrime-thacrylate. The rate of
biosorption was controlled by the rate of diffusion through
the shell of saturated material and a maximum Cu uptake
of ~7 mg g–1 dry wt was observed for a resin–biomass com-
plex containing 8% w/w biomass [38,39]. In P. aeruginosa,
lanthanide biosorption obeyed the Brunauer-Emmett-
Teller isotherm for multilayer adsorption with uptake of
 Bioremedial potential of microbial mechanisms of metal mobilization and immobilization Gadd
La3+, Eu3+ and Yb3+ unaffected by K+, Ca2+, Cl–, SO42–, or
NO3– but strongly inhibited by Al3+ [40].
As well as bacteria, fungi and algae also receive sustained
biosorptive examination. Zn biosorption to Rhizopus
arrhizus was predominantly to wall chitin and chitosan
(maximum sorptive capacity of 312 µmol g–1 dry wt com-
pared to 213 µmol g–1 dry wt for the wall) [31]. Sorption
activity for UO22+, Pb2+, Mn2+, and Co2+ of A. niger struc-
tural cell-wall polysaccharides also depends on the
chitin : glucan ratio [41]. Freeze-dried biomass of white-rot
fungi was found to be the most efficient biomass type for
Cd2+ biosorption [42], whereas dry biomass of Phanerochaete
chrysosporium sorbed mercury and alkylmercury species
with an affinity of CH3HgCl>C2H5HgCl>Hg(II) and max-
imum capacities of 79, 67 and 61 mg g dry wt–1,
respectively [43]. Saccharomyces cerevisiae accumulated toxic
metals when immobilized in a sol–gel matrix with the
entrapment matrix contributing to overall uptake. Affinities
of the biogel for metal accumulation were
Hg>Zn>Pb>Cd>Co, Ni>Cu [35]. Immobilized biomass of
Rhizopus oligosporus was more effective than freely sus-
pended biomass for Cd2+ with a possible contribution to
sorption by the immobilization matrix [34]. A reliable
model based on Fick’s Law of Diffusion and the Langmuir
adsorption model was able to predict experimentally deter-
mined kinetics of Ni2+ removal by fungal biosorbents [44].
Removal of Pb, Cd and Cu by alkali-treated A. niger bio-
mass was greater than removal achieved by granular
activated carbon (F-400) [45]. Benzaldehyde was an effec-
tive fixation agent for Saccharomyces cerevisiae biomass,
which retained its original lead biosorption capacity [46].
For algae, in situ metal removal from the marine environ-
ment has been postulated, with one Chlorella strain
exhibiting maximal removal of 39.4 mg Cd g–1 dry cells
from 50 µM Cd under marine conditions [47]. Cu was pref-
erentially adsorbed to Zn by Cymodocea nodosa, a brown
alga [48]. Little attention has been given to metal–organic
mixtures, though equilibrium uptakes of, for example, Ni
and phenol are changed due to initial pH and other reaction
components [49]. Bacterial, algal and fungal biomass were
able to remove and concentrate gold from solutions in
which it occurred as a gold–cyanide complex [50]. Note that
extracellular polymers are intimately involved in Cd
adsorption by activated sludges [51].
Chemical modification of biomass may create derivatives
with altered metal-binding abilities and affinities. A. niger
mycelium was modified by introducing additional carboxy
or ethyldiamino groups, with the latter substitution
increasing the maximal capacity for nickel from ~70 to
~1060 mmol kg–1. An increase in maximal binding capaci-
ties for Cd, Ni and Zn also resulted in carboxy-modified
biomass (Cd 100–274; Ni 71–182; Zn 185–303 mmol kg–1,
respectively) [52•].
The earlier commercial promise and development of
biosorption appears to have largely ceased and there is no
273
adoption of biosorption as a commercially viable treat-
ment method to date. The lack of commercial
development is somewhat perplexing, although the lack
of specificity and lower robustness of biomass-based sys-
tems compared to ion exchange resins is often cited as a
reason [5••]. Unless such reasons and the reluctance of
industry to adopt and develop biosorption technology are
overcome, the rationale for continued research activity is
weak as few studies provide novel insights or under-
standing of the fundamental physical and chemical
mechanisms involved. This perhaps contrasts with a bio-
geochemical perspective where microbial metal sorption
is contributing to an understanding of metal fate and
cycling in the environment. For example, the application
of surface complexation modeling or a linear program-
ming approach to specific chemical and electrostatic
interactions occurring at the solution–cell-wall interface
can also be applied to biosorption phenomena. Such stud-
ies can relate variations in surface properties with
variations in metal affinity in order to predict metal
mobilities in complex natural systems [53,54,55••,56].
Metal-binding molecules
Eukaryotic metallothioneins and other metal-binding pep-
tides have been expressed in E. coli as fusions to
membrane or membrane-associated proteins such as
LamB, an outer membrane protein that functions as a
coliphage surface receptor and is involved in maltose/mal-
todextrin transport. Such in vivo expression of
metallothioneins provides a means of designing biomass
with specific metal-binding properties [8••,57,58]. Metal-
binding peptides of sequences Gly–His–His–Pro–His–Gly
(HP) and Gly–Cys–Gly–Cys–Pro–Cys–Gly–Cys–Gly (CP)
were engineered into LamB protein and expressed in
E. coli. Cd2+ : HP and Cd2+ : CP stoichiometries were 1:1
and 3:1, respectively, indicating the potency of these pep-
tides for metal binding. Surface display of CP increased
the Cd2+ binding ability of E. coli fourfold. Some competi-
tion of Cu2+ with Cd2+ for HP resulted from strong Cu2+
binding to HP, indicating that the relative metal-binding
affinities of inserted peptides and the wall to metal ion
ratio were important in the design of peptide sequences
and their metal specificities [59••]. Another gene encoding
a de novo peptide sequence containing the metal-binding
motif Cys–Gly–Cys–Cys–Gly was chemically synthesized
and expressed in E. coli as a fusion with the maltose-bind-
ing protein. Such cells possessed enhanced binding of
Cd2+ and Hg2+ compared to cells lacking the peptide [57].
Related to the application of metal-binding molecules is
the identification of genes encoding phytochelatin syn-
thases (PCSs) as phytochelatins play major roles in metal
detoxification in plants and fungi. Heterologous expres-
sion of PCS genes dramatically enhanced metal tolerance,
providing molecular evidence for the role of phytochelatin
molecules in metal tolerance. Future research with
microorganisms and plants may allow testing of the poten-
tial of PCS genes for bioremediation [60•]. A variety
274 Environmental biotechnology
of other metal-binding molecules may have future poten-
tial for metal recovery. Culture filtrates from the
sulfate-reducing bacterium Desulfococcus multivorans
exhibited copper-binding with 12 day old filtrates having
a copper-binding capacity of 3.64 ± 0.33 µmol ml–1 with a
stability constant log10K of 5.68 ± 0.64 (n = 4) [61•]. A hol-
low-fibre reactor containing an engineered E. coli strain
expressing Hg2+ transport and metallothionein accumu-
lated Hg2+ effectively, reducing a concentration of
2 mg L–1 to ~5 µg L–1 [62]. It can be noted that biosen-
sors for the detection of metal bioavailability have been
developed based on the direct interaction between
metal-binding proteins and metal ions. Here, capacitance
changes of the proteins (e.g. a synechococcal metalloth-
ionein and a mercury resistance regulatory protein) were
detected in the presence of femtomolar to millimolar
metal ion concentrations [63•].
Reductive precipitation
Where reduction of a metal to a lower redox state occurs,
mobility and toxicity can be reduced, thus offering
potential bioremediation applications. Such processes
can also accompany other indirect reductive metal pre-
cipitation mechanisms, for example, in sulfate-reducing
bacterial systems where reduction of Cr(VI) can be a
result of indirect reduction by Fe2+ and the produced sul-
fide. Aerobic or anaerobic reduction of Cr(VI) to Cr(III)
is widespread in microorganisms and both ex situ reactor
systems and in situ treatment approaches have been doc-
umented. Problems can occur with Cr(VI) toxicity as well
as maintenance of anaerobic conditions for anaerobic
Cr(VI) reducers. However, several aerobic and anaerobic
Cr(VI) reducers are known with some being able to use
organic contaminants as electron donors for Cr(VI) reduc-
tion, pointing to the potential for in situ treatment of
mixed wastes [3]. U(VI) can be reduced to U(IV) by cer-
tain Fe(III)-dissimilatory microorganisms, for example,
Geobacter metallireducens, and this reduction in solubility
can be the basis of U removal from contaminated waters
and leachates [3]. Sulfur and sulfate-reducing bacteria
are geochemically important in reductive precipitation of
toxic metals, for example, U(VI) and Cr(VI), a process
mediated by multiheme cytochrome c proteins. The
gene encoding cytochrome c7 from Desulfuromonas ace-
tooxidans has been cloned and expressed in Desulfovibrio
desulfuricans, the recombinant organism exhibiting
enhanced expression of metal reductase activities. Such
overproduction of active cytochrome c7 could be impor-
tant in fixed-enzyme reactors or in the production of
organisms with enhanced metal reductase activities for
bioremediation [64]. D. desulfuricans can couple the oxi-
dation of a range of electron donors to reduction of
Tc(VII), mediated by a periplasmic dehydrogenase,
which is precipitated as an insoluble oxide at cell periph-
eries. Resting cells, immobilized in a flowthrough
membrane bioreactor, accumulated substantial quantities
of Tc when supplied with formate as the electron donor,
suggesting the potential of this organism in treatment of
Tc-contaminated wastewater [65••]. In fact, compared to
an E. coli mutant that overexpressed the reductase, the
D. desulfuricans enzyme exhibited a higher specific activ-
ity, reduced anion sensitivity, a better temperature
optimum, and a lower Ks for formate, the electron
donor [66•]. D. desulfuricans can also reduce Pd(II) to cell-
bound Pd(0) with the hydrogen-dependent reduction
being O2-insensitive, thus providing a means of Pd
recovery under oxic conditions [67]. Mention should be
made of those sulfate-reducing bacteria such as
Desulfotomaculum reducens that share physiological proper-
ties with both sulfate- and metal-reducing groups of
bacteria and can grow with Cr(VI), Mn(IV), Fe(III) and
U(IV) as sole electron acceptors [68].
Se(VI) reduction to elemental insoluble Se(0) has been
employed in the remediation of contaminated waters and
soils. Some bacteria can use such a reduction to support
growth, making this a natural process for in situ applica-
tions. Though reduction of oxyanions of As and Se can
occur by different mechanisms, the most environmental-
ly significant process is dissimilatory reduction.
Oxyanions of As and Se can be used in microbial anaero-
bic respiration as terminal electron acceptors providing
enough energy for growth and metabolism. Their reduc-
tion can be coupled to a variety of organic substrates (e.g.
lactate, acetate and aromatics) and the bacteria are found
in a range of habitats and are not confined to any specific
genus. These organisms, and perhaps even the enzymes
themselves, may have applications for bioremediation of
Se- and As-contaminated environments [69••].
Sulfurospirillum barnesii can simultaneously reduce both
NO3– and Se(VI). Kinetic experiments with cell mem-
branes of S. barnesii suggest the presence of constitutive
selenate and nitrate reduction as well as an inducible,
high affinity nitrate reductase, which also has a low affin-
ity for selenate, in nitrate-grown cells. Simultaneous
reduction of micromolar Se(VI) in the presence of mil-
limolar nitrate suggests a role for these organisms in
bioremediating nitrate-rich seleniferous waste-
waters [70••]. It has been shown that selenate does not
sorb to soil components during transport in saturated soil
columns, but rapidly reduces to forms that are strongly
retarded. Such findings suggest that Se will be retained
near the soil surface even under extreme leaching condi-
tions [71]. It should be noted that the reoxidation of Se(0)
is now known to be under microbial control and where
alteration in physico-chemical conditions leads to, for
example, drying out of Se(0)-contaminated sediments or
aerobiosis of subsurface soils, Se(0) can be reoxidized to
form toxic selenite and selenate [72]. The incidental abil-
ity of a variety of microorganisms from all major groups to
reduce Se(VI) and Te(VI) by additional often uncharac-
terized mechanisms offers additional scope for
bioreactor-based approaches [73•]. Finally, rhizosphere
bacteria increased the efficiency of Se and Hg phytore-
mediation by promoting the accumulation of Se and Hg
in tissues of wetland plants [74•].
 Bioremedial potential of microbial mechanisms of metal mobilization and immobilization Gadd
Sulfide precipitation
Sulfate-reducing bacteria (SRB) are anaerobic het-
erotrophs utilizing a range of organic substrates and SO42–
as a terminal electron acceptor. The sulfide produced
from sulfate reduction plays a major role in metal sulfide
immobilization in sediments but has also been applied to
bioremediation of metals in waters and leachates. In one
integrated microbial process for the bioremediation of soil
contaminated with toxic metals, sulfur-oxidizing bacteria
were used to leach metals from contaminated soils by
breakdown of minerals and liberation of acid-labile
sorbed or chemical forms [9••]. A large proportion of the
acidity and almost the entirety of released metals were
removed using bacterial sulfate reduction in an internal
sedimentation reactor. The combination of soil bioleach-
ing followed by separate bioprecipitation of leached
metals by SRB proved to be effective in removing and
concentrating a range of metals, including Zn, Cu and Cd,
from metal-contaminated soil. The resultant solids, once
separated, represented a ~200-fold reduction in volume
compared to the soil, whereas the low toxic metal content
of the clarified liquor produced would permit discharge or
allow recycling of the liquor to the bioleaching stage and
permit water conservation during process operation.
Where soil and water conditions are appropriate, the soil-
leaching component of such an integrated process could
possibly be carried out in situ, although conditions allow-
ing this would be prone to uncertainty. In other cases, a
range of ex situ options, including heap leaching or slurry
bioreactors, could be used to solubilize metals and feed
the bioprecipitation reactor [9••]. Sulfate-reducing bacte-
rial biofilm reactors may offer a means of process
intensification and entrap or precipitate metals, for exam-
ple, Cu and Cd, at the biofilm surface [75••,76]. Mixed
SRB cultures were more effective than pure cultures for
metal removal, which was enhanced by greater produc-
tion of exopolymers in the mixed culture [75••,76].
Anaerobic solid substrate bioreactors have also been used
in laboratory treatment of acid mine drainage with sul-
fate-reducing activity being an important component of
the overall removal process, which exceeded 99% for Cu
and Cd [77]. Note also that a complex array of microbial,
plant and soil interactions, including sulfate reduction,
contribute to metal immobilization and/or removal from
contaminated wetlands [6,78].
Phosphate precipitation
Microbially enhanced chemisorption where biologically
produced phosphate (Pi) precipitates metals as phosphates
is effective for a range of metals as well as radionuclides: a
mixture of accumulative and chemisorptive mechanisms
contribute to the overall process. Zirconium was mineral-
ized by a Citrobacter sp. to a mixture of Zr(HPO4)2 and
hydrated zirconia (ZrO2); biomineralization of uranium as
HUO2PO4 was repressed by zirconium in the presence of
excess Pi. Cell-bound HUO2PO4 facilitated Ni2+ removal
by intercalative ion exchange into the polycrystalline lat-
tice and also promoted Zr removal [79]. Similar
275
phenomena were enhanced in biofilm-immobilized cells
in an air-lift bioreactor; the uranyl phosphate accumulated
by the bacterial biomass has potential as a bio-inorganic
ion exchanger [80]. The Citrobacter acid phosphatase has
been characterized and found to exist in two isoforms (CPI
and CPII) with differing stabilities, which would relate to
their suitability for metal phosphate precipitation [81••]. A
strain of Acinetobacter was isolated from a wastewater-treat-
ment plant operating a biological phosphate removal
process. Polyphosphate was degraded under anaerobic
conditions in the presence of Cd or UO22+, the latter stim-
ulating the reaction that produced free Pi. Cd was
associated with phosphate-containing intracellular gran-
ules [82]. In E. coli, manipulation of polyphosphate
metabolism by overexpression of the genes for polyphos-
phate kinase (ppk) and polyphosphatase (ppx) can result in
lower intracellular polyphosphate, phosphate secretion
and increased metal tolerance [83].
Conclusions
Mechanisms of microbial solubilization and immobiliza-
tion of metal(loid)s, radionuclides and related substances
are of clear potential for bioremediation, with some
processes being integral to the operation of several suc-
cessful in situ and ex situ processes. Although biosorption
research is rather introverted with little recent develop-
ment in an industrial context, work on metal leaching
from contaminated matrices, metal(loid) transformation
and bioprecipitation is leading to ‘field’ development, as
well as providing fundamental scientific insights into
metal–microbe interactions. As well as the biotechnologi-
cal significance, it should be emphasized that this work
also provides understanding of the biogeochemistry of
metal(loid) cycling in the environment and the central
role of microorganisms in affecting metal mobility and
transfer between different biotic and abiotic locations.
Update
Developments of relevance to this topic, published since
submission of the manuscript, have occurred in several
areas including biosorption, alkylation and reduction.
In an attempt to create a metal-capturing system for use in
bioremediation, a peptide library was screened for binding
to ZnO. By engineering a structural component of the fim-
brae of E. coli to display a random peptide library,
metal-chelating bacteria were isolated. Sequences respon-
sible for Zn adherence were identified and characterized
and found to have varying affinities and specificity for
ZnO. Bacterial surface display of such ‘designer’ chelators
in connection with type 1 fimbrae may offer a system for
engineering bacteria with particular biosorptive
properties [84]. Other studies have generated recombinant
strains of Staphylococcus xylosus and Staphylococcus carnosus
with surface-exposed chimeric proteins containing poly-
histidyl peptides for metal ion binding, the first
demonstration using Gram-positive bacteria. Such cells
exhibited enhanced Ni2+ and Cd2+ binding capacity and
276 Environmental biotechnology
may have potential in bioremediation or biosensor applica-
tions [85]. Although the significance of extracellular
polymeric substances in metal biosorption is well known, it
has recently been demonstrated that the composition of
cell-associated polymers and their ability to sorb toxic met-
als are functions of culture media and conditions, which
offers manipulative possibilities for biosorption-type appli-
cations [86]. Citrate-utilizing strains of Pseudomonas
aeruginosa and Pseudomonas putida have been isolated that
can remove Cd, Zn, Cu, Fe, Co and Ni from solution with
incorporation of inorganic phosphate as a precipitant. Such
organisms may have potential in the treatment of metal-
citrate wastes: citrate is used as a complexing agent in
certain decontamination processes [87]. Sulfate-reducing
bacterial strains, isolated from uranium mine tailings, have
been shown to be capable of Al biosorption with Al depo-
sition being exclusively associated with cell surfaces [88].
Further elucidation of the mechanisms of Se methylation
has resulted from monitoring of volatile and dissolved
DMSe and dimethydiselenide, and dimethylselenonium
compounds in soil spiked with selenomethionine. It is sug-
gested that selenomethionine undergoes a methylation
pathway via methylselenomethionine and dimethylselenon-
iopropionate to DMSe, and that oxidation of dissolved
DMSe to dimethylselenoxide may occur in soil [89].
Aqueous phase flux can have an impact on the metabo-
lism of Fe(III)-reducing bacteria, which is important for
predicting their influence on transport and fate of metals
and radionuclides in the subsurface. It has been shown
that removal of biogenic Fe(II) via aqueous phase trans-
port dramatically increased Fe(III) reduction and
associated bacterial growth. As such organisms can
reductively liberate or immobilize metal contaminants,
as mentioned previously, these findings have implica-
tions for understanding the fate of metal, and organic,
pollutants whose geochemical behaviour is linked to
Fe(III) oxide reduction, and the implementation of
bioremediation strategies involving metal-reducing bac-
teria [90]. A diversity of hyperthermophilic microbes can
reduce Fe(III), Mn(IV) and other metals at 100°C.
Reduction was hydrogen-dependent with U(VI) reduced
to U(IV) uraninite, Tc(VII) to Tc(IV) or Tc(V), Cr(VI) to
insoluble Cr(III), and Mn(IV) to Mn(II) with the forma-
tion of manganese carbonate. The metal reduction
activities of these organisms or their enzymes may have
potential for bioremediation of metal-laden water [91]. It
has been shown for the first time that a mesophilic fac-
ultatively anaerobic Fe(III)-reducing bacterium, closely
related to Pantoea (Enterobacter) agglomerans, can couple
the oxidation of acetate or H2 to dissimilatory reduction
of Fe(III), Mr(IV) and Cr(VI) [92]. Some SRB strains
have been shown to reduce arsenate. A Desulfomicrobium
sp. can use arsenate as a terminal electron acceptor in
minimal medium in the absence of SO42–, the mecha-
nism of reduction being a c-type cytochrome or enzyme
associated with such a cytochrome. A Desulfovibrio strain
was unable to use arsenate as a terminal electron accep-
tor but possessed an arsenate reductase [93].
Acknowledgements
The author gratefully acknowledges financial support for his own work
described from the Biotechnology and Biological Sciences Research Council
(SPC 02812, SPC05211, BSW 05375) and the Royal Society.
References and recommended reading
Papers of particular interest, published within the annual period of review,
have been highlighted as:
• of special interest
••of outstanding interest
1. Gadd GM: Fungal production of citric and oxalic acid: importance
•• in metal speciation, physiology and biogeochemical processes.
Adv Microb Physiol 1999, 41:47-92.
This review discusses the physiology and chemistry of citric and oxalic acid
production in fungi and the intimate association of these acids and process-
es with metal speciation, physiology and mobility and their importance and
involvement in key fungal-mediated processes, including metal biogeochem-
istry and biotechnology.
2. Francis AJ: Biotransformation of uranium and other actinides in
radioactive wastes. J Alloys Comp 1998, 271-273:78-84.
3. Lovley DR, Coates JD: Bioremediation of metal contamination.
Curr Opin Biotechnol 1997, 8:285-289.
4. Stephen JR, MacNaughton SJ: Developments in terrestrial bacterial
•• remediation of metals. Curr Opin Biotechnol 1999, 10:230-233.
This review focusses on the bioremediation of mixed wastes and the recent
developments designed to increase metal removal efficiency from contami-
nated sites by, for example, precipitation.
5. Eccles H: Treatment of metal-contaminated wastes: why select a
•• biological process? Trends Biotechnol 1999, 17:462-465.
Key attributes and advantages of microbial systems for metal/radionuclide
bioremediation are compared with proven physico-chemical technologies.
6. Webb JS, McGinness S, Lappin-Scott HM: Metal removal by
sulphate-reducing bacteria from natural and constructed
wetlands. J Appl Microbiol 1998, 84:240-248.
7. Nies DH: Microbial heavy-metal resistance. Appl Microbiol
•• Biotechnol 1999, 51:730-750.
This review details current understanding of known metal resistance sys-
tems in microorganisms, and describes mechanisms of metal ion homeosta-
sis and transport for important toxic metal elements.
8. Chen W, Brühlmann F, Richins RD, Mulchandani A: Engineering of
•• improved microbes and enzymes for bioremediation. Curr Opin
Biotechnol 1999, 10:137-141.
This review emphasises new approaches for genetically engineering microor-
ganisms and enzymes for bioremediation of inorganic and organic pollutants.
9. White C, Sharman AK, Gadd GM: An integrated microbial process
•• for the bioremediation of soil contaminated with toxic metals. Nat
Biotechnol 1998, 16:572-575.
This paper describes an integrated microbiological process for treatment of
metal-contaminated soil. Initial leaching of contaminated soil by sulfur-oxidiz-
ing bacteria was followed by separate precipitation of leachate metals as
insoluble sulfides by a mixed culture of sulfate-reducing bacteria.
10. Seidel H, Ondruschka J, Morgenstern P, Stottmeister U: Bioleaching
of heavy metals from contaminated aquatic sediments using
indigenous sulfur-oxidizing bacteria: a feasibility study. Water Sci
Technol 1998, 37:387-394.
11. Gomez C, Bosecker K: Leaching heavy metals from contaminated
soil by using Thiobacillus ferrooxidans or Thiobacillus thiooxidans.
Geomicrobiol J 1999, 16:233-244.
12. Mercier G, Chartier M, Couillard D, Blais JF: Decontamination of fly
ash and used lime from municipal waste. Environ Management
1999, 24:517-528.
13. Reyes I, Bernier L, Simard RR, Antoun H: Effect of nitrogen source
on the solubilization of different inorganic phosphates by an
isolate of Penicillium rugulosum and two UV-induced mutants.
FEMS Microbiol Ecol 1999, 28:281-290.
14. Reyes I, Bernier L, Simard RR, Tanguay P, Antoun H: Characteristics
of phosphate solubilization by an isolate of a tropical Penicillium
rugulosum and two UV-induced mutants. FEMS Microbiol Ecol
1999, 28:291-295.
 Bioremedial potential of microbial mechanisms of metal mobilization and immobilization Gadd
15. Altomare C, Norvell WA, Björkman T, Harman GE: Solubilization of
phosphates and micronutrients by the plant-growth-promoting
and biocontrol fungus Trichoderma harzianum Rifai 1295-22. Appl
Environ Microbiol 1999, 65:2926-2933.
16. Brandl H, Bosshard R, Wegmann M: Computer-munching
• microbes: metal leaching from electronic scrap by bacteria and
fungi. Process Metallurgy 1999, 9B:569-576.
This paper demonstrates the microbial leaching of metals from electronic
scrap materials.
17. Diels L, DeSmet M, Hooyberghs L, Corbisier P: Heavy metals
•• bioremediation of soil. Mol Biotechnol 1999, 12:149-158.
This paper describes a microbial process for soil bioremediation based on
siderophore-mediated metal solubilization and subsequent metal adsorption
and precipitation by Alcaligenes eutrophus. A large decrease in Cd, Zn and
Pb bioavailability resulted on biotreatment of a sandy soil.
18. Sayer JA, Cotter-Howells JD, Watson C, Hillier S, Gadd GM: Lead
•• mineral transformation by fungi. Curr Biol 1999, 9:691-694.
This work demonstrates the ability of certain soil fungi, for example,
Aspergillus niger, to solubilize and transform the stable lead mineral,
pyromorphite. Such transformation also resulted in the production of
insoluble lead oxalate dihydrate, the first recorded biogenic formation of
this mineral.
19. Gray SN: Fungi as potential bioremediation agents in soil
contaminated with heavy or radioactive metals. Biochem Soc
Trans 1998, 26:666-670.
20. DeGroot RC, Woodward B: Wolfiporia cocos — a potential agent
for composting or bioprocessing Douglas-fir wood treated with
copper-based preservatives. Material Organismen 1998,
32:195-215.
21. Guo L, Frankenberger WT, Jury WA: Adsorption and degradation of
dimethyl selenide in soil. Environ Sci Technol 1999, 33:2934-2938.
22. Stork A, Jury WA, Frankenberger WT: Accelerated volatilization
rates of selenium from different soils. Biol Trace Element Res
1999, 69:217-234.
23. Zhang YQ, Frankenberger WT: Effects of soil moisture, depth, and
organic amendments on selenium volatilization. J Environ Qual
1999, 28:1321-1326.
24. Zhang YQ, Frankenberger WT, Moore JN: Effect of soil moisture on
•• dimethylselenide transport and transformation to non-volatile
selenium. Environ Sci Technol 1999, 33:3415-3420.
This paper determines the influence of soil moisture on the speciation of
organoselenium compounds and provides an explanation for low Se
volatilization rates in flooded soils and sediments.
25. Turpeinen R, PantsarKallio M, Haggblom M, Kairesalo T: Influence of
microbes on the mobilization, toxicity and biomethylation of
arsenic in soil. Sci Total Environ 1999, 236:173-180.
26. Weeger W, Lievremont D, Perret M, Lagarde F, Hubert JC, Leroy M,
Lett MC: Oxidation of arsenite to arsenate by a bacterium
isolated from an aquatic environment. BioMetals 1999,
12:141-149.
27. Bousserrhine N, Gasser UG, Jeanroy E, Berthelin J: Bacterial and
chemical reductive dissolution of Mn-, Co-, Cr-, and Al-substituted
geothites. Geomicrobiol J 1999, 16:245-258.
28. Chang JS, Hwang YP, Fong YM, Lin PJ: Detoxification of mercury by
•• immobilized mercuric reductase. J Chem Technol Biotechnol 1999,
74:965-973.
This paper reports on the successful immobilization and operation of mer-
curic reductase (Hg2+ → Hg0), from a recombinant Escherichia coli, on a
diatomaceous earth support.
29. Butter TJ, Evison LM, Hancock IC, Holland FS: The kinetics of metal
uptake by microbial biomass: implications for the design of a
biosorption reactor. Water Sci Technol 1998, 38:279-286.
30. Prakasham RS, Sheno Merrie J, Sheela R, Saswathi N,
Ramakrishna SV: Biosorption of chromium VI by free and
immobilized Rhizopus arrhizus. Environ Poll 1999, 104:421-427.
31. Zhou JL: Zn biosorption by Rhizopus arrhizus and other fungi. Appl
Microbiol Biotechnol 1999, 51:686-693.
32. Yin PH, Yu QM, Jin B, Ling Z: Biosorption removal of cadmium
from aqueous solution by using pretreated fungal biomass
cultured from starch wastewater. Water Res 1999, 33:1960-1963.
33. Mameri M, Boudries N, Addour L, Belhocine D, Lounici H, Grib H:
Batch zinc biosorption by a bacterial non-living Streptomyces
rimosus biomass. Water Res 1999, 33:1347-1354.
277
34. Aloysius R, Karim MIA, Ariff AB: The mechanisms of cadmium
removal from aqueous solution by nonmetabolizing free and
immobilized live biomass of Rhizopus oligosporus. World J
Microbiol Biotechnol 1999, 15:571-578.
35. Al Saraj M, AbdelLatif MS, El Nahal I, Baraka R: Bioaccumulation of
some hazardous metals by sol-gel entrapped microorganisms.
J Non-Crystalline Solids 1999, 248:137-140.
36. Chang J-S, Chen C-C: Biosorption of lead, copper and cadmium
• with continuous hollow-fiber microfiltration processes. Sep Sci
Technol 1999, 34:1607-1627.
In this work, hollow-fibre crossflow microfiltration using Pseudomonas
aeruginosa was used to biosorb Pb, Cu and Cd. The results lead to the
development of a rapid-equilibrium and a mass-transfer model for single- and
multi-metal biosorption.
37. Kefala MI, Zouboulis AI, Matis KA: Biosorption of cadmium ions by
Actinomycetes and separation by flotation. Environ Poll 1999,
104:283-293.
38. Veglio F, Beolchini F, Boaro M, Lora S, Corain B, Toro L:
Poly(hydroxiethyl methacrylate) resins as supports for copper (II)
biosorption with Arthrobacter sp.: matrix nanomorphology and
sorption performances. Process Biochem 1999, 34:367-373.
39. Veglio F, Beolchini F, Gasbarro A, Toro L: Arthrobacter sp. as a cop-
per biosorbing material: ionic characterisation of the biomass
and its use entrapped in a poly-hema matrix. Chem Biochem Eng
Quart 1999, 13:9-14.
40. Texier A-C, Andres Y, Le Cloirec P: Selective biosorption of lan-
thanide (La, Eu, Yb) ions by Pseudomonas aeruginosa. Environ
Sci Technol 1999, 33:489-495.
41. Tereshina VM, Marin AP, Kosyakov VN, Kozlov VP, Feofilova EP:
Different metal sorption capacities of cell wall polysaccharides of
Aspergillus niger. Appl Biochem Microbiol 1999, 35:389-392.
42. Cihangir N, Saglam N: Removal of cadmium by Pleurotus
sajor-caju basidiomycetes. Acta Biotechnol 1999, 19:171-177.
43. Saglam N, Say R, Denizli A, Patir S, Arica MY: Biosorption of inor-
ganic mercury and alkylmercury species onto Phanerochaete
chrysosporium mycelium. Process Biochem 1999, 34:725-730.
44. Suhasini IP, Sriram G, Asolekar SR, Sureshkumar GK: Nickel
biosorption from aqueous systems: studies on single and multi-
metal equilibria, kinetics, and recovery. Sep Sci Technol 1999,
34:2761-2779.
45. Kapoor A, Viraraghavan T, Cullimore DR: Removal of heavy metals
using the fungus Apergillus niger. Bioresource Technol 1999,
70:95-104.
46. Ashkenazy R, Yannai S, Rahman R, Rabinovitz E, Gottlieb L: Fixation
of spent Saccharomyces cerevisiae biomass for lead sorption.
Appl Microbiol Biotechnol 1999, 52:608-611.
47. Matsunaga T, Takeyama H, Nakao T, Yamazawa A: Screening of
marine microalgae for bioremediation of cadmium-polluted sea-
water. J Biotechnol 1999, 70:33-38.
48. Sanchez A, Ballester A, Blazquez ML, Gonzalez F, Munoz J,
Hammaini A: Biosorption of copper and zinc by Cymodocea
nodosa. FEMS Microbiol Rev 1999, 23:527-536.
49. Aksu Z, Akpinar D, Kabasakal E, Kose B: Simultaneous biosorption
of phenol and nickel (II) from binary mixtures onto dried aerobic
activated sludge. Process Biochem 1999, 35:301-308.
50. Niu H, Volesky B: Characteristics of gold biosorption from cyanide
solution. J Chem Technol Biotechnol 1999, 74:778-784.
51. Guibaud G, Baudu M, Dollet P, Condat ML, Dagot C: Role of extra-
cellular polymers in cadmium adsorption by activated sludges.
Environ Technol 1999, 20:1045-1054.
52. Krämer M, Meisch H-U: New metal-binding ethyldiamino- and
• dicarboxy-products from Aspergillus niger industrial wastes.
BioMetals 1999, 12:241-246.
In this work, fungal biomass was chemically modified with additional carboxy
or ethyldiamino groups, which increased the metal-binding capacity for Zn,
Cd and Ni.
53. Daughney CJ, Fein JB: Sorption of 2,4,6-trichlorophenol by Bacillus
subtilis. Environ Sci Technol 1998, 32:749-752.
54. Daughney CJ, Fein JB, Yee N: A comparison of the
thermodynamics of metal adsorption onto two common bacteria.
Chem Geol 1998, 144:161-176.
278 Environmental biotechnology
55. Small TD, Warren LA, Roden EE, Ferris FG: Sorption of strontium by
•• bacteria, Fe(III) oxide, and bacteria–Fe(III) oxide composites.
Environ Sci Technol 1999, 33:4465-4470.
This paper characterizes the sorption of Sr by bacteria, Fe(III) oxide, and bac-
teria–Fe(III) oxide composites and discusses the importance of these
processes in determining metal mobilities in the environment where hetero-
geneous bacteria–oxide composite solids are commonly found.
56. Cox JS, Smith DS, Warren LA, Ferris FG: Characterizing heteroge-
neous bacterial surface functional groups using discrete affinity
spectra for proton binding. Environ Sci Technol 1999, 33:4515-
4512.
57. Pazirandeh M, Wells BM, Ryan RL: Development of bacterium-
based heavy metal biosorbents: enhanced uptake of cadmium
and mercury by Escherichia coli expressing a metal binding
motif. Appl Environ Microbiol 1998, 64:4068-4072.
58. Valls M, González-Duarte R, Atrian S, De Lorenzo V:
Bioaccumulation of heavy metals with protein fusions of metal-
lothionein to bacterial OMPs. Biochimie 1998, 80:855-861.
59. Kotrba P, Doleková L, de Lorenzo V, Ruml T: Enhanced
•• bioaccumulation of heavy metal ions by bacterial cells due to
surface display of short metal binding peptides. Appl Environ
Microbiol 1999, 65:1092-1098.
This paper describes the engineering and expression of metal binding pep-
tides in E. coli and the resultant increase in Cd2+ binding ability.
60. Clemens S, Kim EJ, Neumann D, Schroeder JI: Tolerance to toxic
• metals by a gene family of phytochelatin synthases from plants
and yeast. EMBO J 1999, 18:3325-3333.
This work identifies the genes encoding phytochelatin synthases thus pro-
viding molecular evidence for the role of phytochelatins in metal tolerance.
61. Bridge TAM, White C, Gadd GM: Extracellular metal-binding
• activity of the sulphate-reducing bacterium Desulfococcus
multivorans. Microbiology 1999, 145:2987-2995.
This paper demonstrates that certain sulfate-reducing bacteria possess an
extracullular metal-binding capacity that is unrelated to the production of sulfide.
62. Chen SL, Kim EK, Shuler ML, Wilson DB: Hg2+ removal by
genetically engineered Escherichia coli in a hollow fiber
bioreactor. Biotechnol Prog 1998, 14:667-671.
63. Corbisier P, van der Leilie D, Borremans B, Provoost A, de Lorenzo V,
• Brown NL, Lloyd JR, Hobman JL, Csoregi E, Johansson G,
Mattiasson B: Whole cell- and protein-based biosensors for the
detection of bioavailable heavy metals in environmental samples.
Analytica Chimica Acta 1999, 387:235-244.
In this work, whole cells containing lux reporter systems, or metal-binding
proteins (e.g. synechococcal metallothionein and a mercury regulatory pro-
tein) were used as biosensors to examine metal bioavailability in environ-
mental samples.
64. Aubert C, Lojou E, Bianco P, Rousset M, Durand M-C, Bruschi M,
Dolla A: The Desulfuromonas acetoxidans triheme cytochrome c7
produced in Desulfovibrio desulfuricans retains its metal reduc-
tase activity. Appl Environ Microbiol 1998, 64:1308-1312.
65. Lloyd JR, Ridley J, Khizniak T, Lyalikova NN, Macaskie LE: Reduction
•• of technetium by Desulfovibrio desulfuricans: biocatalyst
characterization and use in a flow-through bioreactor. Appl
Environ Microbiol 1999, 65:2691-2696.
This paper describes the efficient reduction and precipitation of Tc(VII) by
Desulfovibrio desulfuricans. The metal is precipitated as an insoluble oxide
at cell peripheries.
66. Lloyd JR, Thomas GH, Finlay JA, Cole JA, Macaskie LE: Microbial
• reduction of technetium by Escherichia coli and Desulfovibrio
desulfuricans: enhancement via the use of high activity strains and
effect of process parameters. Biotechnol Bioeng 1999, 66:122-130.
This work compares the Tc(VII) reductive performance of Desulfovibrio
desulfuricans to that of an Escherichia coli mutant strain that overexpresses
the Tc reductase.
67. Lloyd JR, Macaskie LE: Enzymatic recovery of elemental palladium
using sulfate-reducing bacteria. Appl Environ Microbiol 1998,
64:4607-4609.
68. Tebo BM, Obraztsova AY: Sulfate-reducing bacterium grows with
Cr(VI), U(VI), Mn(IV), and Fe(III) as electron acceptors. FEMS
Microbiol Lett 1998, 162:193-198.
69. Stolz JF, Oremland RS: Bacterial respiration of arsenic and
•• selenium. FEMS Microbiol Rev 1999, 23:615-627.
This review highlights some of the recent advances and provides new
insights into the taxonomy, physiology and biochemistry of arsenic- and sele-
nium-respiring bacteria.
70. Oremland RS, Switzer Blum J, Burns Bindi A, Dowdle PR, Herbel M,
•• Stolz JF: Simultaneous reduction of nitrate and selenate by cell
suspensions of selenium-respiring bacteria. Appl Environ
Microbiol 1999, 65:4385-4392.
This paper describes the simultaneous reduction of Se(VI) and nitrate by the
selenium-respiring bacterium Sulfurospirillum barnesii and suggests a pos-
sible role in bioremediation of nitrate-rich seleniferous wastewaters.
71. Guo L, Frankenberger WT, Jury WA: Evaluation of simultaneous
reduction and transport of selenium. Water Resources Res 1999,
35:663-669.
72. Dowdle PR, Oremland RS: Microbial oxidation of elemental seleni-
um in soil slurries and bacterial cultures. Environ Sci Technol
1998, 32:3749-3755.
73. Gharieb MM, Kierans M, Gadd GM: Transformation and tolerance
• of tellurite by filamentous fungi. Mycol Res 1999, 103:299-305.
This paper examines accumulation and transformation of tellurite (TeO32–) by
filamentous fungi. Reduction and precipitation of elemental Te within vac-
uoles and around biomass was demonstrated with only small amounts of Te
being volatilized in a species-dependent process.
74. DeSouza MP, Huang CPA, Chee N, Terry N: Rhizosphere bacteria
• enhance the accumulation of selenium and mercury in wetland
plants. Planta 1999, 209:259-263.
This work demonstrates that rhizosphere bacteria can increase the efficien-
cy of Se and Hg phytoremediation by promoting the accumulation of Se and
Hg in the tissues of wetland plants.
75. White C, Gadd GM: Accumulation and effects of cadmium on
•• sulphate-reducing bacterial biofilms. Microbiology 1998,
144:1407-1415.
This work demonstrates that mixed sulfate-reducing bacterial biofilms are
more effective than pure cultures for metal removal, the mechanisms of accu-
mulation being entrapment and/or precipitation of CdS at the biofilm surface.
76. White C, Gadd GM: Copper accumulation by sulphate-reducing
bacterial biofilms. FEMS Microbiol Lett 2000, 183:313-318.
77. Drury WJ: Treatment of acid mine drainage with anaerobic solid-
substrate reactors. Water Environ Res 1999, 71:1244-1250.
78. O’Sullivan AD, McCabe OM, Murray DA, Otte ML: Wetlands for
rehabilitation of metal mine wastes. Biol Environ Proc Roy Irish
Acad 1999, 99b:11-17.
79. Basnakova G, Macaskie LE: Accumulation of zirconium and nickel
by Citrobacter sp. J Chem Technol Biotechnol 1999, 74:509-514.
80. Finlay JA, Allan VJM, Conner A, Callow ME, Basnakova G, Macaskie
LE: Phosphate release and heavy metal accumulation by biofilm-
immobilized and chemically-coupled cells of a Citrobacter sp.
pre-grown in continuous culture. Biotechnol Bioeng 1999,
63:87-97.
81. Jeong BC, Macaskie LE: Production of two phosphatases by a
•• Citrobacter sp. grown in batch and continuous culture. Enzyme
Microbial Technol 1999, 24:218-224.
This work characterizes the Citrobacter acid phosphatase, which was found
to exist in two isoforms of differing stability and suitability for metal phos-
phate precipitation.
82. Boswell CD, Dick RE, Macaskie LE: The effect of heavy metals and
other environmental conditions on the anaerobic phosphate
metabolism of Acinetobacter johnsonii. Microbiology 1999,
145:1711-1720.
83. Keasling JD, Van Dien SJ, Pramanik J: Engineering polyphosphate
metabolism in Escherichia coli: implications for bioremediation of
inorganic contaminants. Biotechnol Bioeng 1998, 58:231-239.
84. Kjærgaard K, Sørensen JK, Schembri MA, Klemm P: Sequestration
of zinc oxide by fimbrial designer chelators. Appl Environ
Microbiol 2000, 66:10-14.
85. Samuelson P, Wernérus H, Svedberg M, Ståhl S: Staphylococcal
surface display of metal-binding polyhistidyl peptides. Appl
Environ Microbiol 2000, 66:1243-1248.
86. Bonthrone KM, Quarmby J, Hewitt CJ, Allan VJM, Paterson-Beedle M,
Kennedy JF, Macaskie LE: The effect of the growth medium on the
composition of metal binding behaviour of the extracellular
polymeric material of a metal-accumulating Citrobacter sp.
Environ Technol 2000, 21:123-134.
87. Thomas RAP, Beswick AJ, Basnakova G, Moller R, Macaskie LE:
Growth of naturally occurring microbial isolates in metal-citrate
medium and bioremediation of metal-citrate wastes. J Chem
Technol Biotechnol 2000, 75:187-195.
 Bioremedial potential of microbial mechanisms of metal mobilization and immobilization Gadd
88. Hard BC, Walther C, Babel W: Sorption of aluminum by sulfate-
reducing bacteria isolated from uranium mine tailings.
Geomicrobiol J 1999, 16:267-275.
89. Zhang YQ, Frankenberger WT: Formation of dimethylselenonium
compounds in soil. Environ Sci Technol 2000, 34:776-783.
90. Roden EE, Urrutia MM, Mann CJ: Bacterial reductive dissolution of
crystalline Fe(III) oxide in continuous-flow column reactors. Appl
Environ Microbiol 2000, 66:1062-1065.
279
91. Kashefi K, Lovley DR: Reduction of Fe(III), Mn(IV) and toxic metals
at 100°C. Appl Environ Microbiol 2000, 66:1050-1056.
92. Francis CA, Obraztsova AY, Tebo BM: Dissimilatory metal reduction
by the facultative anaerobe Pantoea agglomerans SP1. Appl
Environ Microbiol 2000, 66:543-548.
93. Macy JM, Santini JM, Pauling BV, O’Neill AH, Sly LI: Two new
arsenate/sulfate-reducing bacteria: mechanisms of arsenate
reduction. Arch Microbiol 2000, 173:49-57.