Biotechnology Advances 24 (2006) 427 – 451

www.elsevier.com/locate/biotechadv

Research review paper

Biosorption of heavy metals by Saccharomyces cerevisiae: A review
Jianlong Wang a,b,⁎, Can Chen a
a
Laboratory of Environmental Technology, INET, Tsinghua University, Beijing 100084, PR China
b
State Key Joint Laoboratory of Environment Simulation and Pollution Control, Tsinghua University, Beijing 100084, PR China
Accepted 8 March 2006
Available online 5 June 2006

Abstract

Heavy metal pollution has become one of the most serious environmental problems today. Biosorption, using biomaterials such
as bacteria, fungi, yeast and algae, is regarded as a cost-effective biotechnology for the treatment of high volume and low
concentration complex wastewaters containing heavy metal(s) in the order of 1 to 100 mg/L. Among the promising biosorbents for
heavy metal removal which have been researched during the past decades, Saccharomyces cerevisiae has received increasing
attention due to the unique nature in spite of its mediocre capacity for metal uptake compared with other fungi. S. cerevisiae is
widely used in food and beverage production, is easily cultivated using cheap media, is also a by-product in large quantity as a
waste of the fermentation industry, and is easily manipulated at molecular level.
The state of the art in the field of biosorption of heavy metals by S. cerevisiae not only in China, but also worldwide, is
reviewed in this paper, based on a substantial number of relevant references published recently on the background of biosorption
achievements and development. Characteristics of S. cerevisiae in heavy metal biosorption are extensively discussed. The yeast can
be studied in various forms for different purposes. Metal-binding capacity for various heavy metals by S. cerevisiae under different
conditions is compared. Lead and uranium, for instances, could be removed from dilute solutions more effectively in comparison
with other metals. The yeast biosorption largely depends on parameters such as pH, the ratio of the initial metal ion and initial
biomass concentration, culture conditions, presence of various ligands and competitive metal ions in solution and to a limited
extent on temperature. An assessment of the isotherm equilibrium model, as well as kinetics was performed. The mechanisms of
biosorption are understood only to a limited extent. Elucidation of the mechanism of metal uptake is a real challenge in the field of
biosorption. Various mechanism assumptions of metal uptake by S. cerevisiae are summarized.
© 2006 Elsevier Inc. All rights reserved.

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 428
2. Advantages of S. cerevisiae as biosorbents in metal biosorption. . . . . . . . . . . . . . . . . . . . . . . . . . . . 429
3. Forms of S. cerevisiae in biosorption research . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 430
4. Biosorption capacity of S. cerevisiae . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 430
4.1. Metal ion uptake . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 430
4.2. Biosorption capacity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 431
4.3. Selectivity and competitive biosorption by S. cerevisiae . . . . . . . . . . . . . . . . . . . . . . . . . . . . 432
4.4. Comparison with other biomaterials . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 433

⁎ Corresponding author. Laboratory of Environmental Technology, INET, Tsinghua University, Beijing 100084, PR China. Tel.: +86 10 62784843;
fax: +86 1062771150.

0734-9750/$ - see front matter © 2006 Elsevier Inc. All rights reserved.
doi:10.1016/j.biotechadv.2006.03.001
428 J. Wang, C. Chen / Biotechnology Advances 24 (2006) 427–451

5. Influential factors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 433

5.1. Properties of metal ions in solution . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 434
5.2. Environmental conditions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 434
5.2.1. pH . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 434
5.2.2. Temperature. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 435
5.2.3. Contact time . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 436
5.2.4. Competing ions/co-ions. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 436
5.2.5. Initial concentration of metal ions and biomass . . . . . . . . . . . . . . . . . . . . . . . . . . . 437
5.2.6. Composition of cultural medium . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 437
5.2.7. Cell age . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 438
6. Pretreatment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 439
7. Biosorption equilibrium isotherm models and kinetics models . . . . . . . . . . . . . . . . . . . . . . . . . . . . 439
7.1. Equilibrium isotherm models . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 439
7.2. Kinetics of biosorption. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 440
7.3. Process of metal uptake . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 441
7.4. Kinetic models for S. cerevisiae . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 442
8. Biosorption mechanism by the cell of S. cerevisiae. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 443
8.1. Extracellular accumulation/precipitation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 443
8.2. Cell surface sorption/precipitation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 444
8.3. Intracellular accumulation/ precipitation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 445
9. Instrumental tools and techniques used in metal biosorption studies . . . . . . . . . . . . . . . . . . . . . . . . . 446
10. Discussions and future directions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 447
10.1. Mechanism research . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 447
10.2. Application of biosorption technology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 447
10.3. Screening of biomaterials. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 448
10.4. Hybrid technology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 448
Acknowledgement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 448
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 448

1. Introduction The toxic characteristics of heavy meals are displayed

Heavy metals mentioned in the field of biosorption are
usually classified as the following three categories: toxic
metals (such as Hg, Cr, Pb, Zn, Cu, Ni, Cd, As, Co, Sn,
etc.), precious metals (such as Pd, Pt, Ag, Au, Ru etc.) and
radionuclides (such as U, Th, Ra, Am, etc.), whose
specific weight is usually more than 5.0 g/cm3 (Volesky,
1990a; Bishop, 2002). With the rapid development of
various industries (including mining and smelting of
metalliferous, surface finishing industry, energy and fuel
production, fertilizer and pesticide industry, metallurgy,
iron and steel, electroplating, electrolysis, electro-osmosis,
leatherworking, photography, electric appliance manufac-
turing, metal surface treating, aerospace and atomic
energy installation), wastes containing metals are directly
or indirectly discharged into the environment increasingly,
especially in developing countries, having brought serious
environmental pollution, and threatened biolife (Bishop,
2002; Volesky, 1990a; Wang, 2002a). Moreover, metal as
a kind of resource is becoming rare and rare.
as follows: (1) the toxicity can last for a long time in
nature; (2) some heavy metals even could be transformed
from relevant low toxic species into more toxic forms in a
certain environment, mercury is such a case; (3) the
bioaccumulation and bioaugmentation of heavy metal by
food chain could damage normal physiological activity
and endanger human life finally; (4) metals can only be
transformed and changed in valence and species, but
cannot be degraded by any methods including biotreat-
ment; (5) the toxicity of heavy metals occurs even in low
concentration of about 1.0–10 mg/L. Some strong toxic
metal ions, such as Hg and Cd, are very toxic even in
lower concentration of 0.001–0.1 mg/L (Alkorta et al.,
2004; Volesky, 1990a; Wang, 2002a).
Due to their increasing application and the above
immutable nature, the heavy metal pollution has naturally
become one of the most serious environmental problems
today. Conventional methods for removing metal ions
from aqueous solution have been studied in detail, such as
chemical precipitation, ion exchange, electrochemical
 J. Wang, C. Chen / Biotechnology Advances 24 (2006) 427–451
treatment, membrane technologies, adsorption on acti-
vated carbon etc. However, chemical precipitation and
electrochemical treatment are ineffective, especially when
metal ion concentration in aqueous solution is as low as 1
to 100 mg/L, they also produce large amount of sludge to
be treated with great difficulties. Ion exchange, membrane
technologies and activated carbon adsorption process are
extremely expensive, especially when treating a large
amount of water and wastewater containing heavy metal
in low concentration, so they cannot be used at large scale.
Alternative process is biosorption, which utilizes various
natural materials of biological origin, including bacteria,
fungi, yeast, algae, etc. These biosorbents possess metal-
sequestering properties and can decrease the concentra-
tion of heavy metal ions in solution from ppt to ppb level.
They can effectively sequester dissolved metal ions out of
dilute complex solutions with high efficiency and quickly.
Therefore biosorption is an ideal candidate for the
treatment of high volume and low concentration complex
wastewaters (Veglio and Beolchini, 1997; Volesky,
1990a).
Heavy metal removal by biosorption has been
extensively investigated during the last several decades
(Volesky, 1990a). Some reviews have been published
focusing on different aspects of heavy metal biosorption
(Davis et al., 2003; Gavrilesca, 2004; Kapoor and
Viraraghavan, 1995; Kratochvil and Volesky, 1998;
Malik, 2004; Tsezos, 2001; Volesky, 2001; Volesky and
Holan, 1995; Veglio and Beolchini, 1997; White et al.,
1995). From these reviews, we can see that the research on
biosorption is focused on the following three major fields.
First, the biosorbents. It is necessary to continue to search
for and select the most promising types of biomass from
an extremely large pool of readily available and
inexpensive biomaterials (Kratochvil and Volesky,
1998). Second, the mechanism of biosorption. The
mechanism involved in the metal biosorption is only
understood to a very limited extent to date. It is necessary
to identify the mechanism of metal uptake by biosorbents
and understand microbe–metal interactions. Third, large-
scale experiment. The biosorption process is basically at
the stage of laboratory-scale study. It is in great difficulties
and almost a failure in attempt of applying biosorption
process into practice (Tsezos, 2001). Great efforts should
be taken to improve biosorption process, including
immobilization of biomaterials, improvement of regener-
ation and re-use, optimization of biosorption process.
Some potential biomaterials with high metal-binding
capacity have been identified in part. Among those bio-
sorbents, there are marine algae (e.g. Sargassum natans),
bacteria (e.g. Bacillus subtillis), fungi (e.g. Rhizopus
arrhizus), yeast (e.g. S. cerevisae) and waste microbial
429
biomass from fermentation and food industry. For the
economical reason, researchers have paid much attention
to various by-products from fermentation industry,
because they are produced in large quantities. The
application of these waste microbes as biosorbents for
the biosorption of heavy metals and radionuclides is to kill
two birds with one stone. For it uses waste to dispose
waste. The enterprises can sell their waste biomass and
earn some money, at the same time, they can save the cost
associated with disposing the waste biomass they
produced (Kapoor and Viraraghavan, 1995). S. cerevisiae
is widely used in the food and beverage industry, it is also
a kind of solid waste. Although S. cerevisiae is a mediocre
biosorbent, it is still a concerned biomaterial in biosorp-
tion study because its unique characteristics in compar-
ison with other microorganisms for metal removal (for
details, see the section of “advantages of S. cerevisiae as
biosorbent”). However, few reviews on metal biosorption
by the yeast are published. This review focuses on metal
removal from aqueous solutions by S. cerevisiae based on
a substantial number of references on metal biosorption
and our research work.
2. Advantages of S. cerevisiae as biosorbents in
metal biosorption
Firstly, S. cerevisiae is easy to cultivate at large scale.
The yeast can be easily grown using unsophisticated
fermentation techniques and inexpensive growth media
(Kapoor and Viraraghavan, 1995). Moreover, the yield
of the biomass is also high.
Secondly, the biomass of S. cerevisiae can be obtained
from various food and beverage industries. S. cerevisiae
as a by-product, is easier to get from fermentation
industry, in comparison with other types of waste
microbial biomass. Microorganisms used in enzymatic
industry and pharmaceutical industry are usually involved
in the secret of their products, which makes industries
reluctant to supply the waste biomass. The supply of S.
cerevisiae as waste residuals is basically stable.
Thirdly, S. cerevisiae is generally regarded as safe.
Therefore, biosorbents made from S. cerevisiae can be
easily accepted by the public when applied practically.
Fourthly, but not the last, S. cerevisiae, is an ideal
model organism to identify the mechanism of biosorp-
tion in metal ion removal, especially to investigate the
interactions of metal–microbe at molecular level.
Perego1 and Howell (1997) reported that the use of
yeasts as model systems is particularly attractive because
of the ease of genetic manipulation and the availability of
the complete genomic sequence of S. cerevisiae. In fact,
S. cerevisiae, as a model system in biology, has been
430 J. Wang, C. Chen / Biotechnology Advances 24 (2006) 427–451
explored fully in molecular biology (Zhou, 2002).
Knowledge accumulated on the molecular biology of
the yeast is very helpful to identify the molecular me-
chanism of biosorption in metal ion removal (Eide, 1997,
1998). At the same time, S. cerevisiae can be easily
manipulated genetically and morphologically, which is
helpful to genetically modify the yeast more appropriate
for various purposes of metal removal.
3. Forms of S. cerevisiae in biosorption research
S. cerevisiae in different forms has been studied for
different purposes of research. For example, living cell/
dead cell (Kapoor and Viraraghavan, 1995), intact cell/
deactivated cell, immobilized cell/free cell (Veglio and
Beolchini, 1997), raw material/pretreated cell by
physicochemical process, wild type/mutant cell, floccu-
lent/non-flocculent cell (Marques et al., 1999), engi-
neered/non-engineered cell, lab culture/waste industrial
cell, and cells from different industries (Park et al.,
2003).
Comparing the results of metal biosorption using the
different forms of the yeast can give useful information
for understanding the mechanism of metal uptake by S.
cerevisiae. For example, Ramsay and Gadd (1997), by
examining and comparing the responses of vacuole-
deficient mutants and wild type of S. cerevisiae to
several toxic metals, found that vacuole-deficient strains
are more sensitive to Zn, Mn, Co, Ni with a largely
decreased capacity to accumulate these metals than wild
type, but no change for Cu or Cd. The results confirmed
the essential role of vacuole in detoxification for Zn,
Mn, Co, Ni, but not for Cu and Cd. Immobilization
technique is one of the key elements for the practical
application of biosorption, especially by dead biomass.
Various kinds of immobilized S. cerevisiae have been
studied with different immobilizing materials (Veglio
and Beolchini, 1997). Park et al. (2003) compared two
strains of S. cerevisiae for the biosorption of cadmium.
One strain is ATCC 834 which is used for the production
of l-phenylacetyl carbinol (l-PAC) and another strain,
ATCC 24858 for ethanol production. They found that
the thicker mannan layer and the larger specific surface
layer seemed to benefit a larger cadmium uptake
capacity for the strain S. cerevisiae ATCC 834.
Free cells appear unsuitable in practical application,
largely due to solid/liquid separation problem. However,
Veglio and Beolchini (1997) pointed out that investiga-
tion on the performance of free cells for metal uptake
can provide fundamental information on the equilibrium
of the biosorption process, which is useful for practical
application. Meanwhile, flocculating cell has been
suggested for biosorption, attempting to overcome the
separation problem of free cells (Soares et al., 2002).
Whether to employ living cells or non-living cells for
biosorption is still at arguing stage (Suh and Kim, 2000).
In the early researches on biosorption of heavy metal
ions, living cells were used. However, dead cells have
been found to have the same or even higher uptake
capacity of metal ions, comparing with living cells.
Meanwhile, dead cells can overcome some limits that
living cells are used: nutrition demand, sensitivity to
extreme pH value or higher metal ion concentration, etc.
Therefore, biosorption studies involving dead/pretreated
biomass have dominated during 1980s–90s (Malik,
2004). However, the limitations of the industrial
application of biosorption with immobilized dead cells
have been realized from some pilot plants. For example,
the cost for producing the required biosorbents with
waste biomass was too expensive using immobilized
techniques and using various pre-treatment processes.
Process of regeneration and re-use on-line is complex
and very expensive. For real effluents, the co-existed
ions and organic matters in aqueous solution made
matters even more difficult and more complex. Hybrid
biotechnologies, such as biosorption, bioprecipitation,
and bioaccumulation, using living cells, even together
with physicochemical process, are suggested in recent
years (Malik, 2004; Tsezos, 2001). As for S. cerevisiae,
dead or living cells are the same important in
biosorption studies today. As a waste microbial biomass
from fermentation, study on dead cells of the yeast is
also dominant and necessary. In exploring the mecha-
nism of metal uptake, especially metal–microbe inter-
actions, living cells of S. cerevisiae should be used
inevitably for specific research at molecular level.
4. Biosorption capacity of S. cerevisiae
4.1. Metal ion uptake
A number of references have proved that S.
cerevisiae can remove toxic metals, recover precious
metals and clean radionuclides from aqueous solutions
to various extents. Schott and Gardner (1997) reported
the recovery of light metals, such as aluminium by S.
cerevisiae. Brady et al. (1994f) proved that the yeast
cells of S. cerevisiae treated with hot alkali were capable
of accumulating a wide range of heavy metal cations
(Fe3+, Cu2+, Cr3+, Hg2+, Pb2+, Cd2+, Co2+, Ag+, Ni2+,
and Fe2+). Some typical metals biosorption by S.
cerevisiae are listed in Table 1.
Lead, cadmium, copper, zinc, chromium, nickel,
silver and uranium, etc. have been studied much more
 J. Wang, C. Chen / Biotechnology Advances 24 (2006) 427–451 431

Table 1
Some metal ions studied in S. cerevisiae biosorption
Metal class Metal References
Toxic metals Pb Özer and Özer, 2003; Suh et al., 1999a,b; Goksungur et al., 2005
Cu Wang, 2002b; Bakkaloglu et al., 1998
Zn Bakkaloglu et al. (1998)
Cd Vasudevan et al., 2003; Goksungur et al., 2005; Gomes et al., 2002;
Park et al., 2003
Hg Al-Saraj et al., 1999; Zhu et al., 2004
Co Al-Saraj et al. (1999)
Ni Özer and Özer, 2003; Bakkaloglu et al., 1998
Cr Rapoport and Muter, 1995; Özer and Özer, 2003; Ferraz et al., 2004
As Nguyên-nhu and Knoops (2002)
Precious metals Pd Liu et al., 2003a; Xie et al., 2003a
Pt Xie et al. (2003b)
Au Karamushka and Gadd, 1999; Lin et al., 2005
Ag Bustard and McHale, 1998; Simmons and Singleton, 1996
Radionuclides U, Pu, Am, Ce; Cs, Sr Kedari et al. (2001)
241
Am Liu et al. (2003b)
Sr Avery and Tobin (1992)
U Nakajima, 2001; Popa et al., 2003; Riordan et al., 1997
Se, Sb Pérez-Corona et al. (1997)
U, Sr, Cs Kapoor and Viraraghavan (1997)
Light metal Al Schott and Gardner (1997)

than cobalt, molybdenum, iron, manganese, radium,
selenium, lanthanide, precious metals, etc. It should be
noted that S. cerevisiae can distinguish different metal
species based on their toxicity, such as selenium species
(Se(IV) and Se(VI)), antimony species (Sb(III) and Sb
(V)) and mercury species (CH3Hg and Hg(II)). This
kind of property makes S. cerevisiae useful not only for
the bioremediation, removal or recovery of metal ions,
but also for their analytical measurement (Madrid et al.,
1995; Madrid and Cámara, 1997; Pérez-Corona et al.,
1997).
4.2. Biosorption capacity
The determination of the metal uptake rate by the
biosorbent is often based on the equilibrium state of
sorption system. The sorption uptake rate, q, is usually
expressed in milligrams of metal sorbed per gram of the
(dry) sorbent (the basis for engineering process — mass
balance calculations), or mmol g− 1 or meq g− 1 (when
stoichiometry and/or mechanism are considered) (Kra-
tochvil and Volesky, 1998). Metal ion uptake by S.
cerevisiae has been reported in a substantial number of
references. Table 2 presents some data on the biosorp-
tive capacities of the yeast (in various forms) for
different metal ions.
Based on data presented in Table 2, the magnitude of
metal uptake by S. cerevisiae can be estimated as
follows: for lead, in the order of 2–3, above 10 and less
than 300 mg Pb/g dry weight biomass; for copper, in the
order of 1–2, less than 20 mg Cu/g dry weight yeast; for
zinc, in the order of 1–2, usually less than 30 mg Zn/g
dry weight; for cadmium, in the order of 2–3, usually
above 10 but less than 100 mg Cd/g dry mass; for
mercury, in the order of 2; for chromium and nickel,
usually in the order of 1; for seldom, more than 40 mg/g
dry mass; for precious metals , such as Ag, Pt, Pd, in the
order of 2, around 50 mg/g dry weight yeast.
Biosorptive capacity of radionuclide uranium by S.
cerevisiae is usually between 150 and 300 mg U/g dry
weight biomass. It should be noted that comparative
results from different references involved standardizing
the different ways which the sorption capacity was
expressed. At the same time, uptake rate, q, should be
compared in almost the same equilibrium concentration
of metal in solution, for the purpose of evaluating
performance of the biomaterial (Kratochvil and Volesky,
1998). In particular, there is no standard measurement of
dry weight of biomass, i.e. no standard of dry
temperature and dry hours when drying biomass. Park
et al. (2003) obtained the dry-cell weight by drying cells
at 70 °C until the weight of the cells became constant.
Liu et al. (2002a) measured the dry weight of S.
cerevisiae after drying in a stove for 2 h at 100–120 °C
to calculate the adsorption capacities. Özer and Özer
(2003) dried the yeast at 100 °C for 24 h. Rapoport and
Muter (1995) determined dry weight by drying the
sample at 105 °C until the constant weight was
432 J. Wang, C. Chen / Biotechnology Advances 24 (2006) 427–451

Table 2
Metal uptake by S. cerevisiae (mg metal/g dry weight biomass)
Metal Source or form Uptake capacity a References
Pb Free cells 79.2 Al-Saraj et al. (1999)
Pb Immobilized cellsin a sol–gel matrix 41.9 Al-Saraj et al. (1999)
Pb Whiskey distillery spent wash, lyophilized 189 Bustard and McHale (1998)
Pb Lab cultivated, then dried at 100 °C 270.3 Özer and Özer (2003)
Pb Ethanol-treated waste baker's yeast 17.5 Goksungur et al. (2005)
Cu Adapted and growing cells 2.04–9.05 Donmez and Aksu (1999)
Cu Dry cells from Commercial co. 2.98–12.03 Zhu and Xu (1998)
Cu Waste yeast from fermentation industry and then autoclaved at 120 °C 4.93 Bakkaloglu et al. (1998)
Cu Free cells 6.4 Al-Saraj et al. (1999)
Cu Whiskey distillery spent wash lyophilized 5.7 Bustard and McHale (1998)
Cu Immobilized cells on sepiolite 4.7 Bag et al. (1999)
Cu Waste yeast from brewery, formaldehyde cross-linked cells in column bioreactors 8.1 Zhao and Duncan (1997)
Zn Waste yeast from fermentation industry and then autoclaved at 120 °C 3.45–1.95 Bakkaloglu et al. (1998)
Zn2+ Free cells 23.4 Al-Saraj et al. (1999)
Zn Immobilized cells in a sol–gel matrix 35.3 Al-Saraj et al. (1999)
Zn Whiskey distillery spent wash, lyophilized 16.9 Bustard and McHale (1998)
Zn Immobilized cells on sepiolite 8.37 Bag et al. (1999)
Zn2+ Formaldehyde cross-linked cells in column bioreactors 7.1 Zhao and Duncan (1997)
Cd Deactivated protonated yeast from yeast co. 9.91–86.3 Vasudevan et al. (2003)
Cd Free cell suspended in solution lab culture 35.5–58.4 Park et al. (2003)
Cd Free cell suspended in solution lab culture 14.3–20.0 Park et al. (2003)
Cd Immobilized cells on sepiolite 10.9 Bag et al. (1999)
Cd Waste yeast from brewery, formaldehyde cross-linked cells in column bioreactors 14 Zhao and Duncan (1997)
Cd Ethanol-treated waste baker's yeast 15.6 Goksungur et al. (2005)
Cd Non-living and resting cells from aerobic culture 70 Volesky (1993)
Hg2+ Free cells 64.2 Al-Saraj et al., 1999; Madrid
et al., 1995
Co2+ Free cells 9.9 Al-Saraj et al. (1999)
Ni Waste yeast from fermentation industry and then autoclaved at 120 °C 1.47 Bakkaloglu et al. (1998)
Ni Free cells 8 Al-Saraj et al. (1999)
Ni Lab cultivated, then dried at 100 °C 46.3 Özer and Özer (2003)
Ni Deactivated protonated yeast from yeast co. oven at 80 °C for 24 h 11.4 Padmavathy et al. (2003)
Cr(VI) Lab cultivated, dehydrated at 30 °C, 15% of cell humidity; 80.5% of the viability About 5.5 Rapoport and Muter (1995)
Cr(VI) As a by-product from brewery, formaldehyde cross-linked 6.3 Zhao and Duncan (1998)
cells in fixed-bed column
Cr(VI) Lab cultivated, then dried at 100 °C 32.6 Özer and Özer (2003)
Fe Whiskey distillery spent wash, lyophilized 16.8 Bustard and McHale (1998)
Pd2+ Immobilized cells of waste yeast 40.6 Xie et al. (2003a)
Pt4+ immobilized cells of waste yeast 44 Xie et al. (2003b)
Ag Whiskey distillery spent wash lyophilized 59 Bustard and McHale (1998)
Ag Industrial strain, then lab cultivated and freeze-dried 41.7 Simmons and Singleton (1996)
241
Am Lab cultivated, free cell 7.45–1880.0 b Liu et al. (2002a,b)
U Whiskey distillery spent wash lyophilized 180 Bustard and McHale (1998)
U Beer yeast 8.75 mmol UO2+ 2 /g yeast 2082.5 c Popa et al. (2003)
Washed and unwashed non-viable spent yeast from a company in Greece 360–150 Riordan et al. (1997)
U 150 Tsezos (1997)
Th 63 Tsezos (1997)
a
Metal uptake is not necessarily maximum.
b
Unit: μg U/g.
c
The value calculated by the data 8.75 mmol UO2+
2 /g yeast in Popa et al. (2003).

achieved. Obviously, the numeric value of dry weight of
biomass obtained in different drying conditions is surely
different. Hence, attention should be paid to these
conditions when reading literatures and analysing the
results.
4.3. Selectivity and competitive biosorption by S.
cerevisiae
To determine which metal ions have high affinity
with S. cerevisiae, it is necessary to compare the
 J. Wang, C. Chen / Biotechnology Advances 24 (2006) 427–451
biosorptive capacity of different metal ions under the
same experimental conditions. Also, it has been found
that metal biosorption by S. cerevisia is selective and, in
some cases, competitive. Some relevant work has been
carried out (see Table 3). However, only very limited
information is available on the competitive sorption of
metal ions with fungal biomass (Kapoor and Virar-
aghavan, 1997), especially with the yeast of S.
cerevisiae. In principle, S. cerevisiae has higher affinity
with uranium, lead and mercury than copper, nickel or
other metal ions.
4.4. Comparison with other biomaterials
Table 4 represents a part of comparative results of
metal biosorption capacity between S. cerevisiae and
other biomaterials. Bakkaloglu et al. (1998) investigated
various types of waste biomass including bacteria (S.
rimosus), yeast (S. cerevisiae), fungi (P. chrysogenum),
activated sludge as well as marine algae (F. vesiculosus
and A. nodosum) for biosorption of metals. They
compared the removal efficiency for zinc, copper and
nickel ions at the stage of the biosorption, sedimentation
and desorption. The results showed that S. cerevisiae has
a mediocre efficiency for one- or multi-metal biosorp-
tion systems. By comparing the index qmax of Langmiur
equation with seven types of waste biomass for the
removal of lead ion, Kogej and Pavko (2001) indicated
that lead uptake capacity by S. cerevisiae is in the
middle, in comparison with the other six biomaterials
they used in their study. Vianna et al. (2000) studied the
biosorption capability for Cu, Cd and Zn, using three
kinds of waste biomass from fermentation industries,
Table 3
Comparison of the biosorptive capacity of metal ions (unit: mg/g)
Comparative results
Pb(II)(270.3) N Ni(II)(46.3) N Cr(VI)(32.6)
Hg2+(55.76) N Zn2+(30.27) N Pb2+(24) N Cd2+(20.91) N
Co2+(14.50) = Ni2+(13.50) N Cu2+(4.45)
Pb(189) N U(180) N Ag(59) N Zn(16.9) N= Fe(16.8) N
Cu(5.7) mg/g
Cd2+(14.0) N Cu2+(8.0) = Zn2+(7.1)
Cu(7.01) N Cd(5.50) = Zn(5.01)
Pb2+(60.24) N Cd2+(31.75)
More than 95% sorption of UO2+ 4+
2 , Pu , Am
3+
and Ce3+,
while sorption of Cs+ and Sr2+ were negligible
Cu2+ was significantly affected by presence of Cd2+ and/or Pb2+.
Cd2+ was only slightly affected by presence of both Cu2+ and
Pb2+, and Pb2+ removal was not affected by Cu2+ and/or Cd2+
433
that is, Bacillus lentus, Aspergillus oryzae and S.
cerevisiae. The results showed that protonated B. lentus
had the highest sorption capacity for Cu and Cd,
followed by protonated biomass of A. oryzae and S.
cerevisiae. Donmez and Aksu (1999) studied copper ion
bioaccumulation by adapted and growing cells of S.
cerevisiae, Kluyceromyces marxianus, Schizosacchar-
omyces pombe and Candida sp. They found that the
biosorptive capacity for Cu2+ decreased in the following
order: S. cerevisiae (7.11) N K. Marxianus (6.44) N
Candida sp. (4.80) N S. pombe (1.27). However, the
results indicated that Candida sp. and K. marxianus
were more efficient than S. cerevisiae and S. pombe for
heavy metal resistance and copper(II) bioaccumulation
at higher copper(II) concentrations. Compared with the
excellent biosorbent of fungi Rhizopus for the biosorp-
tion of lead, cadmium, copper, zinc, and uranium, the
common yeast S. cerevisiae is regarded as a ‘mediocre’
metal biosorbent (Volesky, 1994). Kapoor and Virar-
aghavan (1997) found that for the biosorption capacity
of cadmium, S. cerevisiae was higher in comparison
with other adsorbents such as aluminum oxide, activated
carbon, and activated charcoal. Comparing with other
fungi biomaterials, in spite of the mediocre metal
biosorption capacity, S. cerevisiae is a unique biomate-
rial in biosorption research and practical application.
5. Influential factors
Biosorptive capacity is influenced by many factors,
including the status of S. cerevisiae (cell age), properties
of metal ions (radius of ion, valence, etc.) in aqueous
solution, cultural conditions (carbon source, nutrition
Remark References
Mono solute biosorption system, Özer and Özer (2003)
comparison based on Langmiur equation
Mono solute biosorption system, based Al-Saraj et al. (1999)
on the same experimental condition
Mono solute biosorption system Bustard and McHale (1998)
based on Langmiur equation
Mono solute biosorption system, based Zhao and Duncan (1997)
on the same experimental condition
Mono solute biosorption system based on Lu and Wilkins (1996)
Langmiur equation
Mono solute biosorption system based Goksungur et al. (2005)
on Langmiur equation
Mono solute biosorption system based Kedari et al. (2001)
on the similar experimental conditions
Two- or three-metal sorption systems Marques et al. (1999)
(Cu, Pb or Cr), results were compared
with one-solute system
434 J. Wang, C. Chen / Biotechnology Advances 24 (2006) 427–451

Table 4
Compartive study for different biomaterials with S. cerevisiae
Metal Biosorptive capacity (mg metal/g dry weight biomass) References
Zn2+
A.nodosum(25.6) N P. chrysogenum N (19.2) N F. vesiculosus(17.3) N Activated Bakkaloglu et al. (1998)
sludge(9.7) N S. rimosus(6.63) N S. cerevisiae(3.45)
Cu2+ S. rimosus(9.07) N P. chrysogenum(8.62) N F. vesiculosus(7.37) N Activated Bakkaloglu et al. (1998)
sluge(5.54) N S. cerevisiae(4.93) N A. nodosum(4.89)
Ni2+ F. vesiculosus(2.85) N S. rimosus(1.63) N S. cerevisiae(1.47) N A. nodosum(1.11) Bakkaloglu et al. (1998)
Pb2+ Phanerochaete chrysosporium(419.4) N R. nigricans(403.2) N Kogej and Pavko (2001)
M. purpurea(279.5) N S. cerevisiae(211.2) N A. terreus(201.1) N
M. inyoensis(159.2) N Streptomyces clavulgerus(140.2)
Cd2+ or Cu2+ Protonated biomass: Bacillus lentus (≈ 30) N Aspergillus oryzae N S. cerevisiae (b5) Vianna et al. (2000)
Cu2+ Growing cells: S. cerevisiae(7.11) N K. Marxianus(6.44) N Candida sp.(4.80) N S. pombe(1.27). Donmez and Aksu (1999)

supply, composition of growth media, etc.), biosorption
conditions (such as pH, temperature, contact time, co-ions
in solution, initial concentration of metal and biomass,
availability of metal ions and micronutrition etc.).
5.1. Properties of metal ions in solution
Tobin et al. (1984) reported that biosorption of metal
ions by R. arrhizus was linearly influenced by the ionic
radius, but independent of the ionic charge or electro-
static strength (Kapoor and Viraraghavan, 1997).
Pearson classified metallic ions based on a “hardness
scale” defined by their binding strength with F− and I−
under the consideration of the thermodynamics but not
the kinetics (Avery and Tobin, 1993; Remacle, 1990).
Nierboer and Ruchardson proposed a refined classifi-
cation of metals for biological systems in determining
their relativeness, considering the electronegativity,
charge and ionic radius of metals. Hard metals, such
as Na+, K+, Ca2+, Mg2+, are usually nontoxic and often
are essential macronutrients for microbial growth, they
bind preferentially to oxygen-containing (hard) ligands,
such as OH−, HPO42−, CO32−, R–COO−, and _C_O,
whereas soft metals, such as Hg2+, Cd2+ and Pb2+,
which often display greater toxicity, form stable bonds
with nitrogen- or sulfur-containing (soft) ligands, such
as CN−, R–S−, –SH−, NH2− , and imidazol. Borderline or
intermediate metals are less toxic and can even be
detected in certain biomolecules where they assist in
mediating specific biochemical reactions, e.g., Zn2+,
Cu2+ and Co2+ (Avery and Tobin, 1993).
Based on the hard-and-soft principle of acids and
bases, Avery and Tobin (1993) investigated the metal
adsorption characteristics for metabolism-independent
uptake of the metal ions, including Sr2+, Mn2+, Zn2+,
Cu2+, Cd2+, Ti2+ by S. cerevisiae. The results showed that
the complex characteristics of microbial metal uptake
conformed well to the hard-and-soft principle (Avery and
Tobin, 1993).
Biosorption of metals ions such as Sr2+, Mn2+, Zn2+,
Cu , Cd2+ and Pb2+ by freeze-dried R. arrhizus is
2+
2
observed to be related to covalent index (Xm r), where
Xm is electronegativity and r is the ionic radius (Brady
and Tobin, 1995). The greater the covalent index value
of metal ion is, the greater is the potential to form
covalent bonds with biological ligands.
5.2. Environmental conditions
To understand the effects of nutritional and environ-
mental factors on the quality of microbial biomass could
help us to cultivate the desirable types of organisms on
cheap nutrients, and to use it in a suitable form for direct
application (Goyal et al., 2003).
5.2.1. pH
For biosorption of heavy metal ions, pH is one of the
most important environmental factors. The pH value of
solution strongly influences not only the site dissocia-
tion of the biomass' surface, but also the solution
chemistry of the heavy metals: hydrolysis, complexation
by organic and/or inorganic ligands, redox reactions,
precipitation, the speciation and the biosorption avail-
ability of the heavy metals (Esposito et al., 2002; Wang,
2002a: 233–248).
The biosorptive capacity of metal cations increases
with increasing pH of the sorption system, but not in a
liner relationship. On the other hand, too high pH value
can cause precipitation of metal complexes, so it should
be avoided during experiments. For different biosorp-
tion system of metal ions, the optimal pH is different. It
is reported that the optimal pH value is 5–9 for copper
biosorption by S. cerevisiae, and 4–5 for uranium
(Volesky, 1990b: 156). Mapolelo and Torto (2004)
proved that the biosorption capacity of Cd2+, Cr3+, Cr6+,
Cu2+, Pb2+ and Zn2+ is dependent on pH. For all metal
ions they studied, the optimal pH values are all greater
than 5. The optimal pH for Cd and Pb biosorption is 5.8,
 J. Wang, C. Chen / Biotechnology Advances 24 (2006) 427–451
while for Cr(III) and Pb is 5.2. As the pH further
increased, the biosorption capacity subsequently de-
creased. The reason may be that at low pH, the affinity
with the proton at the binding site of yeast is much
greater than that of the metal ion (H+ ≫ M2+), compared
with that at higher pH, where M2+ ≫ H+. Vianna et al.
(2000) obtained a similar conclusion. They found that
the biosorption capacity of metal cations strongly
depends on pH value. For Cu, Cd and Zn, the
biosorption capacity at pH 4.5 is far higher than that at
pH 2.5 and pH 3.5. Electrostatic attraction to negatively
charged functional groups may be one of the specific
biosorption mechanisms. At pH 4.5, the most important
group is phosphate, and the other two main active
molecular groups are carboxyl and sulphate.
Marques et al. (2000) studied the pH effects (i.e.
initial pH value and the pH shift and control) on the
removal of Cu2+ ,Cd2+ and Pb2+ from unbuffered
aqueous solution by non-viable S. cerevisiae (brewery
waste biomass). A pH shift from 4.5–5.0 (optimal pH
range) to a final value of 7.0–8.0 range was observed.
They suggested different removal mechanisms for each
cation: Cu2+ was removed by both the metal sorption
and precipitation due to the pH shift that occured during
the process, while Cd 2+ removal was completely
dependent on this pH shift. Pb2+ was totally and quickly
removed by precipitation in the presence of the yeast
suspension at pH 4.5.
Özer and Özer (2003) found that optimal pH value
for Pb(II) and Ni(II) ion uptake is 5.0. At lower pH, cell
wall ligands were closely associated with the hydronium
ions [H3O+] and restricted the approach of metal cations
as a result of the repulsive force. At higher pH, e.g. 5.0,
divalent positive ions are suitable to interact with
negatively charged groups in biomass. On the other
hand, the outer layer of the cell wall of S. cerevisiae
consists of a coat protein, which can cause a charge
through dissociation of ionisable side groups of the
amino acids. The ionic state of ligands such as carboxyl,
phosphate, imidazole and amino groups will promote
reaction with the positively charged metal ions.
However, metal anions, such as chromium(VI),
exhibit different pH features from metal cations.
Generally speaking, a low pH value is favorable for
biosorption.
The effect of pH on biosorption of Cr(VI) has been
investigated by various researchers using a variety of
different types of biomass. Optimal biosorptive removal
of Cr(VI) at low pH (2.0) has been reported for Rhizopus
nigricans, Bacillus sp. and Dunaliella sp. Özer and Özer
(2003) found that the optimal pH for Cr(VI) biosorption
by S. cerevisiae is 1.0. Anions could be sorbed on the
435
protonated active sites of the biosorbent primarily
electrostatically in nature. At very low pH values, the
surface of sorbent would also be surrounded by the
hydronium ions which enhance the chromium(VI)
interaction with binding sites of the biosorbent by
greater attractive forces. As the pH increased, however,
the overall surface charge on the cells became negative
and decreased the biosorption capacity.
Ferraz and Teixeira (1999) found that the initial pH
adjustment (at pH 5.0) seems to decrease the compet-
itive effect between Pb(II) and Cr(III). In mixed sol-
utions of Pb(II) and Cr(III), biosorption of lead
proceeded faster. This may be due to changes in yeast
binding sites or to different chromium species present in
solution.
The yeast was modified by a cationic surfactant to
improve chromate anions (CrO42−) removal (Bingol et al.,
2004). Zeta potential of modified cells was all positive
and reached a maximum at pH 5.5, which corresponded
to the maximal sorption efficiency for CrO42. Comparing
with the unmodified biomass, the modified biomass not
only increased the maximal removal efficiency of Cr
from 55% to 99.5%, but also shifted the optimal pH from
2 to 5.5. At higher pH values (pH N 7.0), at least four
times more chromium uptake occurred for the modified
cells than unmodified cells (Bingol et al., 2004). This
kind of modification method for the yeast may offer a
promising way to utilize the biomass at high pH for the
removal of metal anions.
The explanation of the pH shift in biosorption
process and the pH effect is helpful to identify the
mechanism of metal biosorption. Jones and Gadd (1990)
discussed the pH effects on ion transport into the yeast
cells at molecular level. More efforts should be devoted
to this problem.
5.2.2. Temperature
Temperature has also an influence on the biosorption
of metal ions, but to a limited extent under a certain
range of temperature, which indicates that ion exchange
mechanism exists in biosorption to some extent.
Biosorption process is usually not operated at high
temperature because it will increase the operational cost
(Wang, 2002a).
Brady and Duncan (1994b) found that temperature
(5–40 °C) had minor effect on the accumulation level of
Cu2+, Co2+ or Cd2+ by free cells of S. cerevisiae in
suspension.
Adsorption reactions are normally exothermic, so
biosorption capacity increases with decrease of temperature
(Kapoor and Viraraghavan, 1997). In the range of 15–
40 °C, the maximum equilibrium biosorption capacity for
436 J. Wang, C. Chen / Biotechnology Advances 24 (2006) 427–451
Pb(II), Ni(II) and Cr(VI) ions by the inactive S. cerevisiae
was reached at temperature of 25 °C. The decrease in
capacity at higher temperature between 25 and 40 °C
revealed that the processes of biosorption for these metal
ions by S. cerevisia are exothermic. The decrease of
biosorption capacity at higher temperature may be due to
the damage of active binding sites in the biomass (Özer and
Özer, 2003). However, Goyal et al. (2003) found that the
metal biosorption of Cr(VI) by S. cerevisiae increases with
increasing temperature in the range of 25–45 °C, they
explained that higher temperature would lead to higher
affinity of sites for metal or binding sites on the yeast. The
energy of the system facilitates Cr(VI) attachment on the
cell surface to some extent. When the temperature is too
high, there is a decrease in metal sorption due to distortion
of some sites of the cell surface available for metal
biosorption.
Also, Zhao and Duncan (1998) have found that the
regeneration of the saturated yeast (immobilized S.
cerevisiae by formaldehyde) is satisfying at 4 °C by the
combination of reduction and desorption using formal-
dehyde and HNO3.
5.2.3. Contact time
The biosorption process of heavy metal by S.
cerevisiae usually completes rapidly. The biosorption
of metals such as copper, zinc, lead and uranium by non-
growing cells of S. cerevisiae is a rapid process and
often reaches equilibrium within several hours (Kapoor
and Viraraghavan, 1997). The biosorption of Pu, Am
and Ce with immobilized cells of S. cerevisiae reached
equilibrium within 60 min. In the case of U, equilibrium
attained in 100 min (Das et al., 2002).
Generally speaking, the biosorption capacity and the
removal efficiency of metal ions by S. cerevisiae
became higher with prolonging the contact time. How-
ever, in practice, it is necessary to optimize the contact
time, considering the efficiency of desorption and
regeneration of the biomass. Ferraz et al. (2004)
optimised the sorption time for Cr(III) by S. cerevisiae
from a brewery company in the sorption–desorption
process, the result showed that a 30 min sorption period
was the best option to ensure the metal removal from
solution and good recovery from biosorbent.
The biosorption of metals discussed in a number of
references was mainly through the passive mode.
However, Malik (2004) pointed out that short biosorption
studies have low possibility to observe and assess the
delayed intracellular accumulation and hence, most of
such studies concluded with the surface adsorption of the
metal. Moreover, the studies, monitoring metal removal
by growing cells, often realize the metabolically linked
intracellular accumulation. Recent studies have proved
that in spite of low apparent growth rate, growing cells are
able to remove metal ions continuously through internal
detoxification mechanisms (Malik, 2004). The active
mode can contribute significantly to metal ion removal for
yeast (Kapoor and Viraraghavan, 1997). It is meaningful
to carry out research on the application of growing cells of
yeast in bioremediation. In a word, equilibrium time of
active cells of yeast in solution needs more research in the
future.
5.2.4. Competing ions/co-ions
Real industrial effluent usually contains various ionic
components, including metal cations and anions. Some
studies indicated that cations and anions additional to
the ions of interest have a generally detrimental impact
on metal accumulation (Suh and Kim, 2000).
Usually, the biosorption capacity of one metal ion is
interfered and reduced by co-ions, including other metal
ions and anions presenting in solution, however the
gross uptake capacity of all metals in solutions remains
almost unchangeable.
Competitive effect occurred in a mixed solution
containing lead and chromium during the biosorption
process by flocculating brewer's yeast (Ferraz and
Teixeira, 1999). The yeast of S. cerevisiae seems to have
more affinity, higher selectivity and biosorption capacity
to Pb(II) than to Cr(III) in aqueous solutions.
Goksungur et al. (2005) observed that the compet-
itive biosorption capacities of the ethanol-treated yeast
for all metal ions of Pb, Cu and Cd were lower than that
under the non-competitive conditions. The decrease of
metal uptake in competitive conditions was thought to
be a response to increased competition between same
charged species for binding sites of the ethanol-treated
yeast cells. The order of the sorption capacity was found
as follows: Pb N Cu N Cd for both pH 4.0 and 5.0.
Pb2+ accumulation by S. cerevisiae was seriously
inhibited by Hg2+. Uptake capacity of Pb2+ decreased
(from 0.22 to 0.02 mmol Pb2+/g cell dry weigh in the
presence of Hg2+), but the total amount of accumulated
metals was not changed (Suh and Kim, 2000).
Goyal et al. (2003) also found that the biosorptive
capacity of Cr(VI) ion by dried S. cerevisiae at the end
of 24 h was significantly reduced by lower concentra-
tion of Fe(III), whereas this concentration of Fe(III) did
not appreciably affect the ultimate uptake of Cr(VI) by
S. equisimilis and A. niger.
As for radionuclides, the influence of co-ions on the
biosorption capacity is not obvious. Liu et al. (2002a)
reported that there were no significant differences for the
biosorption of 241Am by S. cerevisiae in the mixed
 J. Wang, C. Chen / Biotechnology Advances 24 (2006) 427–451
solution containing Au3+ or Ag+, even at 2000 times
higher concentration than 241Am concentration. Similar
results were obtained by Das et al. (2002). Accumulation
of long-lived radionuclides such as 233U, 239Pu, 241Am by
immobilized S. cerevisiae was also investigated in
presence of the cationic impurities such as Al, Be, Cd,
Cr, Fe, Mn, Pb, Ce, Dy, Eu, Gd and Sm. The results
showed that there were no significant differences.
Light metal ions, such as Ca2+, Na+, K+, are present
in industrial wastewater. The experimental data has
shown that those light metal ions have little effect on
heavy metal biosorption, which indicates that the
affinity of the yeast for those light metal ions is far
less than for heavy metal ions. Alkali metal ions and
alkaline earth metals were not absorbed by the yeast,
possibly because they lack the ability to form complexes
with the various ligand groups present on the fungal
surface (Wang, 2002a; Kapoor and Viraraghavan, 1997;
Brady and Duncan, 1994c).
The presence of anions also affects the biosorption of
metal ions. Kapoor and Viraraghavan (1997) reported
that the biosorption capacity decreased in the presence
of ethylenediamine tetraacetate (EDTA), sulphate,
chloride, phosphate, carbonate, glutamate, citrate and
pyrophosphate. The anions existing in the solution may
form a complex with the metal ions, which would reduce
the metal biosorption capacity seriously. However, Das
et al. (2002) found that anionic impurities such as Cl−,
NO3−, SO42−, C2O42− and CH3COO− do not have any
adverse influence on Pu4+ sorption, while the presence
of PO34− reduces the sorption by the yeast S. cerevisiae
(the effect of anions was studied separately up to 0.5 mol
l− 1 of their individual concentrations) (Das et al., 2002).
Nowadays only limited information is available on
the competition of fungal biosorption, including S.
cerevisiae. More research is required in competitive
study for the biosorption systems containing two- or
multi-metal ions by utilizing multi-parameter mathe-
matical models (Wang, 2002a).
5.2.5. Initial concentration of metal ions and biomass
The uptake rate of the metal ion will increase
along with increasing the initial concentration if the
amount of biomass is kept unchanged. Contrary to
that, biosorptive capacity of the metal ions is
inversely proportional to the initial concentration of
the biomass when the initial concentration of metal
ions is kept constant. Increase of the biomass
concentration of the biosorption system could result
in increasing the sorption site interactions. When the
biomass concentration is low, metal ions in the
solution would not only be adsorbed to the surface
437
of the biomass, but also enter into intracellular part
through facilitating the concentration gradient of metal
ion (Wang, 2002a).
As a matter of fact, biosorptive capacity of metal ions
was reported to be related to the ratio of the concentration
of initial metal ions to the concentration of biomass.
Vasudevan et al. (2003) found that equilibrium uptake
for cadmium(II) ion by deactivated protonated yeast was
directly proportional to the ratio of the initial metal ion
concentration to the sorbent mass. Therefore, both
aspects cannot be neglected when assessing the influence
of concentration of the metal ion and biomass on
biosorption, otherwise, error would occur (Schiewer and
Volesky, 1995a).
5.2.6. Composition of cultural medium
5.2.6.1. Glucose. Glucose is a carbon source widely
used. It also acts as an important source of energy. For the
biosorption of living cells, the addition of glucose and
other trace elements into a biosorption system will
enhance the growth of living cells, and facilitate metal
biosorption. Some researches were carried out on the role
of glucose in metal ion biosorption. Stoll and Duncan
(1996) investigated the removal of Cu2+, Cr6+, Cd2+, Ni2
+
and Zn2+ from electroplating effluent by living cells of
S. cerevisiae. The results showed that pretreatment of the
yeast cells with glucose increased the amount of metal
removed, whilst direct addition of glucose into the yeast–
effluent mixed solution had no effect on the amount of
metal accumulated. Avery and Tobin (1992) reported a
different result. In their experiment, glucose was added
into the living yeast cells solution just 5 min before the
addition of Sr2+, the presence of glucose resulted in a
stimulation of Sr2+ uptake due to metabolism-dependent
intracellular Sr2+ accumulation, mainly in the vacuole.
Mapolelo and Torto (2004) also reported that the
pretreatment of the S. cerevisiae by using 10–20 mM
glucose increased the removal efficiency for Cd2+, Cr3+,
Cu2+, Pb2+ and Zn2+ by 30–40%, but the pretreatment
by using 60 mM glucose decreased the removal of Cr6+
by almost 50%. The mechanism of Cr6+ uptake may be
different from other metal ions. Higher uptake of Cr(III)
may be related to the glucose tolerance factor (GTF), a
cationic Cr 3+ complex of lower molecular weight
consisting of Cr3+, nicotinic acid, and the amino acids
glycine, glutamic acid and cysteine. It is believed that
GTF is mainly connected with carbohydrate metabolism
by yeast cells. Carbohydrate metabolism by S. cerevi-
siae, requiring Cr(III) as a component of GTF, con-
tributed to such a high uptake of Cr(III) in the presence
of glucose (Mapolelo and Torto, 2004).
438 J. Wang, C. Chen / Biotechnology Advances 24 (2006) 427–451
In order to compare S. cerevisiae with other yeast cells,
Gomes et al. (2002) investigated five strains of Rhodotor-
ula mucilaginosa for accumulating free and complexed
silver ion by metabolism-dependent and independent
processes. Silver dicyanide [Ag(CN)2−] uptake was
strictly dependent on glucose. Ag+ uptake by both live
and dead biomass was studied. The addition of glucose
decreased the Ag+ uptake rate by living UFMG-Y02,
Y27, and Y35 cells by 16% to 25%, while an improved
uptake rate of Ag+ (115% and 13%, respectively) by
strains CBS 316 and UFMG-Y01 was observed. It seems
that the role of glucose in biosorption depends on the type
of strains and the status of metal ions (free or complex),
even for the same biomass and for the same metal ions.
A strain of Trichoderma atroviride, isolated from
sewage sludge, was found to be capable of surviving at
high concentrations of heavy metal ions (copper, zinc and
cadmium). Lín and Vazquez (2003) found that when the
mycelia were transferred to medium without dextrose, T.
atroviride was capable of uptaking more metal ion than in
the presence of dextrose. The lowest uptake of metal ions
in the presence of glucose, and the highest values of
metals removal were achieved for autolysed mycelia
(Errasquín and Vázquez, 2003). They proposed two
possible and alternative explanations for the above results.
Firstly, an active detoxification process would occur in the
presence of an abundant source of carbon such as glucose,
therefore reducing the amount of metal ion in the cell.
Secondly, no carbon source available will trigger the cell
autolysis. Cell autolysis will not only increase surface area
of the autolysed cell wall, but also lead to exposure of
intracellular binding sites of the biomass, thus providing
access to additional negatively charged binding sites.
Therefore, physical binding of metal ions by cell surfaces
rather than an active process is the main uptake
mechanism, in which, fungal cell wall plays an important
role in metal biosorption. Also, Chojnacka et al. (2004)
found that the decrease in initial glucose concentration
favored synthesis of cells, hence elevating the biosorption
uptake of cations by microalgae Spirulina sp. For
Chryseomonas sp. MGF-48, a bacterium isolated from
electroplating effluent, uranium uptake by washed
bacterial cells (198.2 mg/g dry weight, under starvation
conditions for 16 h) was reduced by the addition of
glucose but remained higher than (156.5 mg/g) that
observed for cells not subject to starvation (151 mg/g)
(Malekzadeh et al., 2002). Uranium uptake in the
presence of selected carbohydrates decreased as follows:
xylose N arabinose N mannose N maltose N glucose.
5.2.6.2. Other compositions of cultural medium. Goyal
et al. (2003) found that supplement of cysteine, glucose,
ammonium sulphate, phosphate and ammonium chloride
in the fermentation media for the growth of S. cerevisiae
increased the Cr(VI) uptake. The best recovery of Cr(VI)
was obtained when the cells were grown in cysteine-
supplemented medium, followed by glucose-, phos-
phate-, ammonium sulphate-, and ammonium chloride-
rich media. Different nutrients clearly led to different
functional groups in the corresponding cell surface.
Cysteine inserts –S and –N ligands, glucose C-ligands,
ammonium N-ligands and phosphates P-ligands (Engl
and Kunz, 1995). All of these, cysteine-rich media gave
the highest adsorption capacity. Simmons and Singleton
(1996) have also obtained similar results. They found
that addition of L-cysteine into the growth medium
increased the biosorption capacity for silver, and also
increased the content of protein and sulphydryl group in
the freeze-dried and viable yeast cells. However,
increasing the concentration of L-cysteine (from 1 to
5 mM) resulted in decreasing the cell numbers in the
medium, in comparison with the control test without
adding L-cysteine. Precipitates on exposed cells to silver
ion were analysed by TEM with X-ray microanalysis, the
peak occurred due to silver or sulphur, indicating that the
silver ion may have bound to sulphur-containing
molecules and precipitated around them.
Dostaleka et al. (2004) investigated the biosorption
of Cd2+, Cu2+ and Ag+ ions by C-, N-, P-, S-, Mg- and
K-limited cells of S. cerevisiae. The binding capacity of
yeast cells for cadmium decreases in the following order:
K-limited ≥ Mg-limited ⋍ C-limited N N-limited ⋍ S-
limited N P-limited. For Ag+ ions: P-limited N K-limit-
ed N C-limited ≥ N-limited ⋍ Mg-limited N S-limited. For
copper ion: K-limited N Mg-limited ≥ C-limited N N-lim-
ited ⋍ P-limited N S-limited.
5.2.7. Cell age
Cell age of biomass has also an influence on metal
biosorption (Goyal et al., 2003; Kapoor and Virar-
aghavan, 1997). Usually, the cells at lag phase or early
stages of growth have a higher biosorptive capacity
for metal ions than that of stationary phase (Kapoor
and Viraraghavan, 1997). Simmons and Singleton
(1996) reported that Ag+ uptake capacity by younger
cells (24 h old) of an industrial strain of S. cerevisiae
was almost twice of that older cells (96 h old) (0.187
and 0.387 mmol Ag+/g dry mass, respectively). Their
work also proved that intracellular components bind
more Ag+ than the cell wall. The addition of sulphur-
containing amino acids into the growth medium
improved the silver biosorption capacity by increasing
cell protein levels and/or altering levels of sulphur
containing compounds within the cell.
 J. Wang, C. Chen / Biotechnology Advances 24 (2006) 427–451
6. Pretreatment
Yeast cells killed by extreme chemical and physical
conditions may also show very different properties for
metal accumulation, compared with the original yeast (Lu
and Wilkins, 1996). Now various pretreatment methods are
reported to deal with the yeast cells of S. cerevisiae.
Physical methods include vacuum and freeze-drying,
boiling or heat, autoclaving, and mechanical disruption.
Chemical methods include treatment with various organic
and inorganic reagents, such as acid and caustic, methanol,
formaldehyde, etc. Those methods are found to improve
metal biosorption to some extent. Alkali treatment of
fungal biomass has increased the metal uptake capacity
significantly, whereas acid treatment of biomass almost has
no influence on metal biosorption (Wang, 2002a; Wang et
al., 2000; Kapoor and Viraraghavan, 1997).
Because of the important role of cell wall in the metal
biosorption by non-viable cells, metal biosorption may be
enhanced by heat or chemical sterilization or by crushing.
Thus degraded cells would offer a larger available surface
area and expose the intracellular components and more
surface binding sites due to the destruction of the cell
membranes (Errasquín and Vázquez, 2003).
The biosorption of cadmium and lead ions from
synthetic aqueous solutions using yeast biomass was
investigated (Goksungur et al., 2005).The waste baker's
yeast cells were treated with caustic, ethanol and heat
methods, and the highest metal uptake values for Ca2+
and Pb2+ were obtained by ethanol-treated yeast cells.
However, Suh and Kim (2000) obtained the different
results on pretreatment. The equilibrium uptake capacity
of lead (mg Pb2+/g cell dry weight) decreased in the
following order: original cell (260) N 5 times autoclaved
cell for 15 min (150) N grinded cell after drying (100) N
autoclaved cell for 5 min (30).
7. Biosorption equilibrium isotherm models and
kinetics models
The assessment of a solid–liquid sorption system is
usually based on two types of investigations: equilib-
rium batch sorption tests and dynamic continuous-flow
sorption studies (Volesky and Holan, 1995).
Several reviews on the equilibrium models and
kinetics models can be found in a number of references
(Gavrilesca, 2004; Kratochvil and Volesky, 1998; Vol-
esky, 2003; Veglio and Beolchini, 1997). Volesky, in his
book entitled “sorption and biosorption” (http://www.
Biosorption.mcgill.ca), also offered a detailed introduc-
tion to biosorption equilibrium and kinetics, e.g. single-
sorbate isotherms, multi-sorbate sorption equilibrium
439
(multi-component Langmiur models considering electro-
static binding, the effect of pH, surface complex model,
Donnan model considering ionic strength, Wilson model
for ion exchange etc.), biosorption batch dynamics (mass
transfer model for biosorption rate), dynamic continuous-
flow reactor/contactor systems modelling of column
performance including equilibrium column model, mass
transfer model and derivation of mass transfer model to
evaluate column sorption performance.
7.1. Equilibrium isotherm models
Equilibrium isotherm models are usually classified
into the empirical equations and mechanistic models,
based on the mechanism of metal ion biosorption.
Mechanistic models can be used not only to represent,
but also to explain and predict the experimental
behavior (Liu et al., 2002b; Pagnanelli et al., 2002).
The empirical models for single solute systems used
to describe the biosorption equilibrium are Langmuir,
Freundlich and Brunauer–Emmett–Teller (BET) mod-
els. Langmuir model (L type, based on monolayer ad-
sorption of solute) and Freundlich model (F type,
developed for heterogeneous surfaces) are the most
widely accepted and used in a number of references. The
BET model describes the multi-layer adsorption at the
adsorbent surface, and assumes that a Langmuir iso-
therm applies to each layer. These models can provide
information of metal-uptake capacities and differences
in metal uptake between various species (Kapoor and
Viraraghavan, 1995; Pagnanelli et al., 2002; Volesky
and Holan, 1995). Scatchard plots, which originally
describe the attractions of proteins for small molecules
and ions, were developed to describe the biosorption
equilibrium (Kapoor and Viraraghavan, 1995; Wang et
al., 2000; Wang, 2002a). The total number of binding
sites may be extrapolated from Scatchard plots if
complete saturation of the biomass binding sites is not
observed (Kapoor and Viraraghavan, 1995). To describe
two- or multi-metal ion biosorption systems, various
extended Langmiur models (also called comparative
Langmiur model) or Freundlich type models have been
developed. More details or examples on those empirical
models can be referred to the relevant references (Aksu
et al., 1997; Chong and Volesky, 1995; Kapoor and
Viraraghavan, 1995; Liu et al., 2002b; Pagnanelli et al.,
2002; Volesky and Holan, 1995).
However, those empirical models can hardly explain
the biosorption behaviour with a meaningful physical
interpretation, and predictive conclusions cannot be drawn
for systems operating under different conditions. The
mechanistic conclusions from the good fit of the models
440 J. Wang, C. Chen / Biotechnology Advances 24 (2006) 427–451
alone should be avoided. Moreover, biosorption isotherms
may exhibit an irregular pattern due to the complex nature
of both the biomaterials and its varied multiple active sites,
as well as the complex solution chemistry of some metallic
compounds (Volesky and Holan, 1995; Kapoor and
Viraraghavan, 1995; Kim et al., 1998).
Therefore, mechanical models have been proposed
not only to represent experimental behaviour but also to
explain and predict biosorption behaviour based on the
understanding of biosorption mechanisms. Two kinds of
mechanistic models are ion exchange model and surface
complexation model.
A research group led by Volesky at McGill
University has carried out a series of work on modelling
biosorption, considering metal biosorption mechanisms
involving ion exchange and/or complexation (Chong
and Volesky, 1995; Figueira et al., 2000; Schiewer and
Volesky, 1995a,b, 1996, 1997; Volesky, 2003; Yang and
Volesky, 1999). Uranium uptake was strongly pH
dependent by a non-living protonated Sargassum
fluitans seaweed biomass. Based on the mechanism of
ion exchange between protons in the biomass and
hydrolysed uranium ion species, a mathematical model
was developed for predicting biosorption isotherm of
uranium at different pH values (Yang and Volesky,
1999). Figueira et al. (2000) also found that the ion
exchange model was suitable for the cadmium-seaweed
biomass system.
However, Pagnanelli et al. (2002) pointed out that the
ion exchange model assumed that the free site
concentration was always the same, but in the case of
weakly acidic site, this quantity changed with pH.
Metal biosorption by non-living biomaterials was
involved in the complexation between metal ions and
various functional groups presenting on the surface of
the biomass cell wall, such as carboxyl, phosphate,
hydroxyl etc. Hence, surface complexation models have
been applied to quantify metal adsorption by bacterial or
algae surfaces (Kim et al., 1998; Volesky, 2003; Liu et
al., 2002b). Fein et al. (1997) utilized acid/base titrations
to determine deprotonation constants for the important
surface functional groups, i.e. carboxyl, phosphate, and
hydroxyl sites displayed on the cell wall surface. Site-
specific stability constants for the important metal–
bacteria surface complexes were also obtained through
the metal–bacteria adsorption experiments using Cd,
Cu, Pb, and Al. The results showed that cell wall func-
tional groups display a high affinity for aqueous metals.
This approach can quantitatively assess the effects of
bacteria on metal adsorption in a wide range of near-
surface fluid–rock systems (Fein et al., 1997). Kim et al.
(1998) used a surface complexation model to simulate
lead biosorption by a kind of marine brown algae,
considering adsorption phenomena as chemical reac-
tions in solution, especially metal ion-surface complex-
ation mechanism. A surface active site and equilibrium
constants for binding parameters were determined by
titration. The model can predict the pH effect and
explain the effects of multi-ion and the other competent
chemicals such as EDTA.
The theory of surface coordination chemistry, widely
used in micro interface of solid–liquid of particles in
aqueous solution (Tang, 2000), is becoming an important
quantitative computing method for the adsorption
process. However, how to use the surface complexation
models based on the theory of surface coordination
chemistry to describe the metal biosorption, a few
references are available.
As for the biomaterial of the yeast S. cerevisiae, the
classical Langmiur model and Freundlich model have
been employed to describe single metal biosorption
system, and in most cases, both models fitted the
experimental data very well (correlation coefficient was
usually larger than 0.99).
Ting and Sun (2000) successfully employed the
Langmiur model and the Freundlich model to simulate
copper uptake by the non-viable immobilized cells of S.
cerevisiae. However, some researches indicated that the
simple Langmuir model is not applicable for metal-
biosorption systems (Vasudevan et al., 2003).
Table 5 gives a part of biosorption isothermal
constants for Langmiur model and Freundlich model
reported in the references. From Table 5, we can see that
in most cases, classical Langmiur model and Freundlich
model can represent single metal biosorption behaviour
for various forms of S. cerevisiae. The mechanistic
model or empirical competitive model was also used to
model multi-metal or competitive biosorption systems
in the case of the biomass of S. cerevisiae. However, the
biosorbents relevant to these works are mainly bacteria
and algae, not S. cerevisiae.
In conclusion, researches on co-ions biosorption
models are only at their preliminary stage. How to
utilize mathematical model with multi-parameters to
describe competitive biosorption is a future direction of
biosorption research (Wang et al., 2000).
7.2. Kinetics of biosorption
Kinetics studies and dynamic continuous-flow inves-
tigations, offering information on the rate of metal
uptake, together with the hydrodynamic parameters, are
very important for biosorption process design (Volesky
and Holan, 1995). However, biosorption kinetics studies
 J. Wang, C. Chen / Biotechnology Advances 24 (2006) 427–451 441

Table 5
Isothermal constants of Langmiur model and Freundlich model for metal–S. cerevisiae system
Form of S. cerevisiae Metal Langmiur model Freundlich model References
2 2
qmax b r KF 1/n r
mg g− 1 1 mg− 1
Untreated Cu(II) 1.25 0.34 0.97 Wang (2002b)
Methanol-treated Cu(II) 0.99 0.31 0.96 Wang (2002b)
Formaldehyde-treated Cu(II) 0.75 0.22 0.98 Wang (2002b)
Glutaraldehyde-treated Cu(II) 2.4 0.2 0.85 Wang (2002b)
Dried at 100 °C for 24 h Pb(II) 270.3 0.0155 0.98 Özer and Özer (2003)
Dried at 100 °C for 24 h Ni(II) 46.3 0.0436 0.96 Özer and Özer (2003)
Dried at 100 °C for 24 h Cr(VI) 32.6 0.0478 0.98 Özer and Özer (2003)
Free cell Cd 132.5 0.0027 0.98 Park et al. (2003)
Free cell Cd 125 0.0012 0.98 Park et al. (2003)
Intact and free cell Am 0.41 0.996 0.999 Liu et al. (2002a)
Ethanol-treated waste yeast Cd2+ 31.75 0.092 0.998 3.08 0.69 0.967 Goksungur et al. (2005)
Ethanol-treated waste yeast Pb2+ 60.24 0.0661 0.9986 3.865 0.84 0.99 Goksungur et al. (2005)
Cationic surfactant-modified Cr(VI) 94.34 0.41 0.98 31.15 0.31 0.97 Bingol et al. (2004)
Lab cultivated, free cell Cr(VI) 6.08 0.5 Goyal et al. (2003)
Flocculating brewer's yeast Cr(III) 13.26 0.0704 0.998 Goyal et al. (2003)
Flocculating brewer's yeast Pb(II) 1.072 0.33 0.898 Ferraz et al. (2004)
Deactivated protonated Ni(II) 11.4 0.0315 0.993 Padmavathy et al. (2003)
Protonated cell 0.1 N HCl 3 h For Cu, Zn, Cd, Langmuir transformations were nonlinear Vianna et al. (2000)
Langmiur model: q = qmax b Ce / (1 + bCe), where qmax (mg/g) is the maximum metal specific uptake and b is the Langmuir constant (l/mg), ratio of the
adsorption/desorption rates, Ce is the metal equilibrium concentration (mg/l), q the metal specific uptake q (mg of metal/g of biomass).
Freundlich model: qe = KFc1/n
e , where, K is a biosorption equilibrium constant, representative of the uptake capacity, and n is a constant indicative of
biosorption intensity.

are insufficient according to the references published so
far (Wang, 2002a).
7.3. Process of metal uptake
Generally speaking, the metal biosorption process by
living cells is regarded as a two-step process. First, metal
ions are adsorbed to the surface of cells by interactions
between metal–functional groups displayed on the
surface of cells, such as carboxyl, phosphate, hydroxyl,
amino, sulphur, sulphide, thilo functional group, etc. The
first step, also called passive biosorption, is metabolism-
independent and proceeds rapidly within several minutes
by any one or a combination of the following metal-
binding mechanisms: coordination, complexation, ion
exchange, physical adsorption (e.g. electrostatic) or
inorganic microprecipitation. Passive biosorption is a
dynamic equilibrium of reversible adsorption–desorp-
tion. Metal ions bound on the surface can be eluted by
other ions, chelating agent or acid. The second step,
metal ions penetrate the cell membrane and enter into the
cells, also called active biosorption. Metal uptake by
non-living cells is mainly in the passive mode (Volesky,
1990a,b; Madrid and Cámara, 1997; Veglio and
Beolchini, 1997; Wang et al., 2000). By investigating
the biosorption of Cr(VI) and Fe(III) on Streptococcus
equisimilis, S. cerevisiae and Aspergillus niger, Goyal et
al. (2003) confirmed that the metal uptake by micro-
organisms occurred in two stages: passive uptake which
takes place immediately, and active uptake which takes
place slowly. The first stage is thought to be physical
adsorption or ion exchange at the cell surface, reaching
the adsorption equilibrium within 30–40 min at the end
of rapid physical adsorption. Ferraz et al. (2004)
observed the similar phenomenon in Cr(III) uptake by
S. cerevisiae over a period of 24 h.
Suh et al. (1998a,b) investigated the process of Pb2+
accumulation in the cells of S. cerevisiae by TEM
(Transmission Electron Microscope) technology. The
results showed that the biosorption process for Pb2+ was
dependent upon the initial concentration of the metal ion
and biomass in unbuffered aqueous solution. The time to
reach an equilibrium state was significantly shortened
from 96 to 24 h as the cell dry weight increased from
0.56 to 5.18 g/l. With a cell concentration of 1 g/L and
initial Pb2+ concentration of 100 mg/L, the first step,
metabolism-independent one, was a rapid process, in
which Pb2+ binds to the cell wall within 3–5 min. In the
second step, Pb2+ penetrated through the cell membrane
and entered into the cytoplasm, but this step cannot be
clearly labelled as metabolism-dependent or -independent.
Third step was Pb2+ accumulation in the cell cytoplasm
442 J. Wang, C. Chen / Biotechnology Advances 24 (2006) 427–451
after 24 h, which was obviously independent on meta-
bolism even though the cells have already entered a dead
phase.
Vasudevan et al. (2003) also found that the process of
cadmium biosorption by inactive and protonated cells of
S. cerevisiae was dependent on the availability of metal
ion in aqueous solution or metal ion concentration.
However, the researchers suggested that the adsorption
process occurred in four distinct steps and the rates for
these steps decreased sequentially. The rate of cadmium
uptake in each case was pseudo-second-order, with
respect to the metal ion concentration (Vasudevan et al.,
2003).
Therefore, the process of metal ion biosorption was
related to the ratio of initial metal ion concentration to
sorbent. Hence, assessing biosorption process of metal
ion must be carefully done.
7.4. Kinetic models for S. cerevisiae
Kinetic models can be helpful to better understand
the mechanisms of metal biosorption and to evaluate
performance of biosorbents for metal removal. Gavri-
lesca (2004) developed the models to determine both the
required number of binding sites for metal ions and the
rate of adsorption under various environmental condi-
tions, using a batch reactor mass balance and the
Langmuir theory of adsorption to surfaces of continuous
dynamic systems. Volesky (http://www.biosorption.
mcgill.ca/) offers a detailed summarization on how to
model dynamic continuous-flow reactor/contactor sys-
tems and evaluate column performance. These sorption
column models include equilibrium column model
(ECM), mass transfer model (MTM) and derivation of
mass transfer model. However, this review only
introduced some examples on biosorption kinetics,
especially related to the biomass of S. cerevisiae.
Pseudo-first-order and pseudo-second-order equations
were used to fit the experimental data of Ni(II) uptake by
the deactivated protonated yeast of S. cerevisiae (Padmav-
athy et al., 2003). In comparison with the adsorption rate
constants of both the models, we can see that the second-
order model provided the best correlation of the data. The
correlation coefficient for the second-order kinetics at all Ni
(II) concentrations was greater than 0.97. The values of qe
increased from 2.23 to 9.31 (mg/g), as the initial
concentrations of nickel(II) varied from 10 to 200 mg/l.
The values of rate constants k2 (g/mg min) decreased from
2.15 × 10− 2 to 8.27 ×10− 3 and initial sorption rate k (mg/g
min) increased from 0.717 to 0.107, as the initial nickel(II)
concentration (C0) increased from 10 to 200 mg/l
(Padmavathy et al., 2003).
Singh et al. (2001) reported a similar results in the
study of Ni(II) and Cr(VI) sorption kinetics by
Microcystis in a single and multi-metallic system:
pseudo-second-order equation provided a more appro-
priate description than the first-order equation. More-
over, the initial sorption rate was not proportional to
the initial concentration and intraparticle diffusion rate
parameters were not directly related to initial concen-
tration, suggesting that neither external mass transfer
nor intraparticle diffusion were rate-limiting factors.
Thus, external mass transfer and intraparticle diffusion
together were involved in the sorption process.
According to Madrid et al. (1995), methylated
mercury CH3Hg+ was completely absorbed within 2–
3 min, and no saturation phenomenon was observed. The
reason may be that the yeast was able to rapidly reduce
CH3Hg + to other more volatile mercury species.
However, Hg(II) was accumulated very slowly and
only about 20% had been captured after 3 h. The process
of the Hg(II) uptake followed Michaelis–Menten
kinetics for the period of 24 h, i.e. having larger but
saturable binding sites or transport channels.
Vasudevan et al. (2003) investigated the kinetics of
biosorption of cadmium(II) ions by deactivated pro-
tonated yeast converted to sodium from over different
stages of sorption at varying solution concentrations
and sorbent dosages. Each step followed a pseudo-
second-order rate kinetic. The results showed the
adsorption process of cadmium(II) ions displayed in
four distinct steps. The initial process of external mass
transfer was fast within a few minutes, termed as the
first stage of sorption. The second, third and fourth
stages of sorption were found to be clearly separated by
a plateau, depending on the concentration or availabil-
ity of metal ions in the solutions. Vasudevan et al.
(2003) obtained the initial rate of the first stage by
considering liquid–solid mass transfer coefficient
(initial sorption rate is from 0.1120 min− 1 at initial
metal concentrations of 10 mg/l to 0.0455 min− 1 at
initial metal concentration of 100 mg/l). The research-
ers calculated the sorption rate of the second, the third
and the fourth stage, according to the pseudo-second-
order model. The rates for these steps decreased
sequentially and related to the ratio of metal ion
concentration to sorbent. From the viewpoint of
application, the second stage with low contact time
was likely to be useful.
Based on the mechanism of reduction of Cr(VI) at
pH b 2, Park et al. (2005) suggested a simple rate
equation as follows:
d½CrðVIÞ=dt ¼ −k½CrðVIÞ½OC
 J. Wang, C. Chen / Biotechnology Advances 24 (2006) 427–451
where: OC represents the equivalent organic compound
capable of reducing Cr(VI) (mmol/l), k represents the
rate of reduction.
The equation could fit the Cr(VI) biosorption by dead
fungal biomass (such as A. niger, Rhizopus oryzae, S.
cerevisiae and Penicillium chrysogenum), which sup-
ports the mechanism that Cr(VI) was removed via a
redox reaction (Park et al., 2005).
Various metal-biomass biosorption systems have
been reported to follow the pseudo-second-order equa-
tion, for example, lead (Pb2+) by Rhodotorula auran-
tiaca (Cho et al., 2005), Cu(II) ions by algae Cladophora
crispate (Özer et al., 2004), and Cr(VI) by Mucor
hiemalis (Tewari et al., 2005), which indicates that the
external mass transfer limitations in the system can be
neglected and the chemical sorption is the rate-limiting
step (Özer et al., 2004).
Dodic et al. (2001) utilized the KEKAM (Kolmo-
gorov–Erofeev–Kozeeva–Avrami–Mampel) equation
to describe the kinetics of zinc biosorption by the living
cells of S. cerevisiae (mean correlation coefficient is
0.96). KEKAM model, a global kinetic equation,
developed by a group of Russian scientists, can describe
the topochemical reactions. Topochemical reactions are
localized (Dodic et al., 2001).
8. Biosorption mechanism by the cell of S. cerevisiae
The mechanism of metal biosorption is complicated
and not fully understood. The status of biomass (living
or non-living), types of biomaterials, properties of
metal-solution chemistry, ambient/environmental con-
ditions such as pH, will all influence the mechanism of
metal biosorption. In the last few years, some reviews
have been published focusing on different aspects of
biosorption mechanism, such as physical–chemical
mechanism, metal detoxification, transfer mechanism
and molecular biology development (White et al.,
1995; White and Gadd, 1995; Lovley and Coatest,
1997; Rosen, 2002; Eide, 1997; Perego1 and Howell,
1997; Wang and Yang, 1996). Two types of metal
sequestering are passive mode by dead or inactive cells
of S. cerevisiae and active mode by living cells.
Passive mode is independent of energy, mainly through
chemical functional groups of the material, comprising
the cell and particularly cell wall. Active mode is
metabolism-dependent and related to the metal trans-
port and deposition. Of course, passive metal uptake
may occur when the cell is metabolically active
(Volesky, 1990b). In this review, the mechanisms of
metal biosorption will be discussed according to the
location where the metal removed from the solution:
443
extracellular accumulation/precipitation, cell surface
sorption/precipitation, intracellular/accumulation (Veglio
and Beolchini, 1997).
8.1. Extracellular accumulation/precipitation
Some prokaryotic (bacteria, Archaea) and eukary-
otic (algae, fungi) microorganisms can produce or
excrete exracellular polymeric substances (EPS), such
as polysaccharides, glucoprotein, lipopolysaccharide,
soluble peptide etc. These substances possess a
substantial quantity of anion functional groups which
can adsorb metal ions. References published on metal
biosorption with EPS mainly focus on the bacterial
organism, such as Bacillus megaterium, Acinetobacter,
Pseudomonas aeruginosa, sulphate-reducing bacteria
(SRB), Cyanobateria or activated sludge (Liu et al.,
2001), whereas EPS study for fungi and algae is
limited (Flemming and Wingender, 2001; Wang and
Yang, 1996; Pirog, 1997).
Suh et al. (1999b) investigated the effect of EPS on
Pb2+ removal by a polymorphic fungus Aureobasidium
pullulans. The results showed that Pb2+ only accumu-
lated on the surface of the intact cells of A. pullulans due
to the existence of EPS, whereas Pb2+ penetrated into
the inner cellular parts of the EPS-extracted cells of A.
pullulans. The longer the storage of cells, the higher the
uptake capacity of Pb2+ by intact cells due to the
increase in the amount of excreted EPS. More than 90%
of the Pb2+ removal was due to excreted EPS, based on
maximal Pb2+ accumulation amount. The biosorptive
capacity of Pb2+ by the EPS-extracted cells was much
less than that of the intact cells and remained constant,
irrespective of the storage time. Suh et al. (1998b) also
discovered that initial rate of Pb2+ uptake by live cells of
S. cerevisiae is lower than that of dead cells, while in the
case of A. pullulans, both the capacity and the initial rate
of Pb2+ accumulation in the live cells are higher than
those in the dead cells, due to the presence of EPS for
live A. pullulans.
The roles of EPS on metal removal in a biosorption
system are usually neglected or ignored, especially in the
case of fungi and yeast. Among the limited studies on
metal removal by EPS, most of them are related to the
EPS extracted from intact organism cells, but not the EPS
in living cells. Although conspicuous extracellular layers
are mainly associated with bacterial cells, whether the
yeast of S. cerevisiae excretes EPS is unclear. Suh et al.
(1998b) implied that the strain of S. cerevisiae used in
their experiment did not excrete EPS. However, floc-
culent strain of S. cerevisiae has been suggested to be
used in metal biosorption due to higher uptake capacity
444 J. Wang, C. Chen / Biotechnology Advances 24 (2006) 427–451
than that of non-flocculent strain and other unique traits
(Soares et al., 2002). The mechanism of flocculation is
believed to be related to a specific protein on the surface
of cell wall, i.e. lectin. Proteins on the surface of the yeast
cells could be extracted by EDTA. With respect to that
feature, lectin seems to be considered as a kind of EPS.
Flocculation of the yeast may vary significantly, even
disappear under certain conditions.
8.2. Cell surface sorption/precipitation
The cell wall tends to be the first cellular structure to
come in contact with metal ions, excluding a possible
existing extracelluar layer mainly related to bacterial
cells. Two basic mechanisms of metal uptake by cell
wall are as follows: stoichiometric interaction between
functional groups of cell wall composition, including
phosphate, carboxyl, amine as well as phosphodiester;
and physicochemical inorganic deposition via adsorp-
tion or inorganic precipitation. Nowadays, complexa-
tion, ion exchange, adsorption (by electrostatic interaction
or van der Waals force), inorganic microprecipitation,
oxidation and/or reduction have been proposed to explain
metal uptake by organism (Volesky, 1990a,b; Liu et al.,
2002b).
The role of separate cell part in the removal of metals
was investigated in the 1990s. Brady and Duncan (1994a)
found that the blocking of amino, carboxyl, or hydroxyl
groups of isolated cell walls from the yeast S. cerevisiae
reduced the uptake capacity of Cu2+, indicating that these
groups play a role in the binding of Cu2+, and implying
that both the protein and the carbohydrate fractions of the
cell walls are involved in the binding of heavy metal
cations. Wang (2002b) also confirmed that the carboxyl
and amino group in the cell wall played an important role
in Cu2+ removal by the waste yeast cells of S. cerevisiae
modified by methanol and formaldehyde. Brady et al.
(1994g) obtained partially purified products (glucan,
mannan, and chitin, respectively) from the isolated cell
walls of the yeast S. cerevisiae by chemical enzymatic
methods. Metal biosorption by the isolated components
revealed that each component accumulated greater
quantities of the cations than the intact cell wall. The
yeast cell wall without the protein component (removed
by pronase) decreased the metal accumulation by 29.5%,
indicating that protein is involved in the heavy metals
removal, such as cobalt, copper and cadmium. The data
also showed that the outer mannan–protein layer of the
yeast cell wall is more important than the inner glucan–
chitin layer in the accumulation of heavy metal cation.
Simmons and Singleton (1996) also confirmed that the
sulphydryl groups of these amino acid residuals are major
metal-binding components of the protein. However, they
pointed out that an isolated cell wall contributes little for
Ag+ removal, in comparison to the whole cell.
Many evidences have proved that ion exchange
mechanism exists in biosorption system (Schneider et
al., 2001; Veglio and Beolchini, 1997). Rapid release of
70% of cellular K+, followed by a slower release of
approximately 60% of cellular Mg2+, but little loss of
Ca 2+ , was observed in Cu2+ accumulation by S.
cerevisiae (Brady and Duncan, 1994c), indicating the
existence of an ion exchange mechanism.
According to Vasudevan et al. (2002), biosorption of
monovalent ions, such as Na+ and K+ by deactivated
protonated yeast of S. cerevisiae was accompanied by
H+ release, whereas biosorption of divalent ions, such as
Ca2+ and Mg2+ was sorbed not only by proton dis-
placement, but also by additional mode, which was not
accompanied by the release of H+. The total maximal
biosorptive capacity of divalent ions was higher than
that of the monovalent ions. The release of Ca2+, Mg2+
or H+ was also observed in the biosorption process of
heavy metal ions (Sr2+, Mn2+, Zn2+, Cd2+, Cu2+, Ti+) by
living, non-metabolising cells of S. cerevisiae, which
also confirmed the existence of ion exchange.
Is ion exchange the main mechanism in metal
biosorption removal? Brady and Tobin (1995) found
that the total ions displaced (H+ + Mg2+ + Ca2+) ac-
counted for only a small portion of the metal ions taken
up in the biosorption of metal ions (Sr2+, Mn2+, Zn2+,
Cd2+, Cu2+, Pb2+) by freeze-dried R. arrhizus. This
indicates that ion exchange is neither the sole nor the
main mechanism for metal biosorption by fungi. How-
ever, Davis et al. (2003) believed the ion exchange was
the main mechanism for metal ion uptake by brown
algae. In particular, they thought the term ion exchange
was rather an umbrella term to describe the experimental
observations, probably the precise binding mechanism
range from physical binding (i.e. electrostatic or
London–van der Waals forces) to chemical binding
(i.e. ionic and covalent).
Therefore, the definition of ion exchange should be
clear before discussing the importance of its role in
biosorption.
Veglio and Beolchini (1997) observed that complex-
ation was involved in metal removal. Davis et al. (2003)
defined complexation or coordination as combination of
cations (often called as central atom) with molecules or
anions containing free electron pairs (bases, often called
as the ligand (s)). A multidentate ligand contains more
than one ligand atom which can be responsible for
combining the metal cation(s). Avery and Tobin (1993)
applied the hard-and-soft principle of acids and bases to
 J. Wang, C. Chen / Biotechnology Advances 24 (2006) 427–451
explore the interactions between metal ions with living,
non-metabolising cells of S. cerevisiae for metabolism-
independent biosorption. Ca2+, Mg2+ or H+ displace-
ment was observed in the biosorption process. The
release of Ca2+ and Mg2+ (indication of ionic binding of
the metal) and H+ displacement (indication of covalent
binding of the metal) seemed to be clearly dependent on
the metal concentration. Brady and Tobin (1995)
observed that Ca2+ and Mg2+ were released for each
tested ions, whereas H+ displacement occurred for the
borderline tested ions only. They concluded that hard
metal Sr2+ adsorption was only due to ionic binding and
the borderline ions (Mn2+, Zn2+, Cd2+, Cu2+, Pb2+)
exhibited a significant degree of covalent binding.
Precipitation and redox reactions in the cell surface
are also reported by many researchers. A research group
at Xiamen University in China has carried out a series of
experiments to explore the removal mechanism of
precious metal ions by dead cells of S. cerevisiae using
various modern instrumental tools and surface technol-
ogy. They found that precious metal ions, such as Pd2+
(Liu et al., 2003a; Xie et al., 2003a), Pt4+ (Xie et al.,
2003b), Au3+ (Lin et al., 2005), Ag+ (Lin et al., 2001)
and Rh3+ (Lin et al., 2001), were unexceptionally bound
to the cell wall of the yeast and then in situ reduced
into the corresponding metal particle. Taking Au3+ as
an example, Lin et al. (2005) studied the mechanism of
Au3+ biosorption by utilizing XRD, FTIR and XPS
techniques. The results demonstrated that the Au3+
bound on the cell wall of the biomass was reduced to
Au0, where Au3+ binding sites may be the hydroxyl and
the carboxylate anions, and the aldehyde group of the
reducing sugar may serve as the electron donor.
Biosorption often involves in many kinds of mechan-
isms. Kratochvil et al. (1998) proved that the maximal
uptake of Cr(VI) by protonated Sargassum biomass at
pH 2 was due to simultaneous anion exchange and the
reduction of Cr(VI) to Cr(III).
8.3. Intracellular accumulation/ precipitation
When the extracellular concentration of metal ions
was higher than that of intracellular, metal ions could
penetrate into the cell across the cell wall and
membrane of the biomass by free diffusion. Metal
ions can also enter into the cell if the cell wall was
disrupted by natural force (e.g. autolysis) or artificial
force (mechanical force or alkali treatment etc.). The
above process is independent of metabolism. However,
the process of intracellular accumulation/precipitation
discussed here mainly relates to the living cells of
biomass, and is an energy-driven process and depen-
445
dent on active metabolism. Metal ions transported
across the cell membrane, are transformed into other
species or precipitated within the cell by active cells,
including transportation.
In the past decades, a large number of references on
metal ion transport in the baker's yeast S. cerevisiae
have been published (Eide, 1997, 1998; Portnoy et al.,
2001). Baker's yeast has been found to possess two or
more substrate-specific transport systems for accumu-
lating any single metal ion. When the metal ion was in
short supply, the activity of high-affinity systems of S.
cerevisiae is limited. In contrast, one or more low-
affinity systems play the predominant roles when the
metal ion was abundant. Recent genetic studies have
identified the transcriptional and post-transcriptional
regulatory mechanisms in mediating metal ion uptake
systems of S. cerevisiae. Also a large number of yeast
genes that function in metal ion transport (including
intracellular transport) or its regulation have been con-
firmed (Eide, 1997, 1998). This review will not discuss
these research findings about metal transport on S.
cerevisiae and more details can be attached in other
reviews (Eide, 1997, 1998).
After entering into the cell, the metal ions are
compartmentalized into different subcellular organelles
(e.g. mitochondria, vacuole etc.). Vijver et al. (2004)
summarized the metal ion accumulation strategies, es-
pecially internal compartmentalization strategies. Per-
ego1 and Howell (1997) reviewed the identified genes
and mechanisms of the controlling sensitivity to toxic
metal ions for both the budding yeast S. cerevisiae and
the fission yeast Schizosaccharomyces pombe. They
discussed the influencing factors (e.g. cellular thiols)
and relevant genes on transport and the products of
genes directly and indirectly involved in the transport or
sequestration of the metal ions. Metal accumulation
strategies for essential and non-essential metal ions may
be different. For essential metals, limiting metal uptake
or strategies with active excretion, storage in an inert
form and/or excretion of stored metal are the main
strategies. For non-essential metals, excretion from the
metal excess pool and internal storage are the major
strategies and metal concentration of the cells will in-
crease with the elevating external concentration (Vijver
et al., 2004). They pointed out that the cellular
sequestration mechanisms mainly have two types: the
formation of distinct inclusion bodies and binding
metals to heat-stable proteins. The former includes
three types of granules: type A, amorphous deposits of
calcium phosphates, e.g. Zn; type B, mainly containing
acid phosphatase, accumulating Cd, Cu, Hg and Ag; and
type C, excess iron stored in granules as haemosiderin.
446 J. Wang, C. Chen / Biotechnology Advances 24 (2006) 427–451
The latter mechanism mainly relates to a specific metal-
binding protein, metallothioneins (MT), which has a low
molecular weight and is cysteine-rich, usually occurring
in the animal kingdom, plants, eukaryotic microorgan-
isms and some prokaryotes. MT can be induced by
many substances, including heavy metal ions, such as
Cd, Cu, Hg, Co, Zn etc. (Vijver et al., 2004). However,
MT in the yeast of S. cerevisiae can only be induced by
Cu, hence called Cu-MT. MT has received much
attention recently because of the potential application
in metal removal (Wang and Yang, 1996). In addition to
MT, other cellular thiols influencing the sensitivity to
toxic metals include glutathione (GSH), phytochelatins
(≡ cadystins (γ-Glu-Cys)nGly), and labile sulfide (Per-
ego1 and Howell, 1997; Gharieb and Gadd, 2004). Tri-
peptide glutathione (GSH) is a typical low molecular
weight cellular thiol and functions as a storage form of
endogenous sulphur and nitrogen as well as detoxification
of metal ions. GSH in S. cerevisiae may account for 1% of
the cell dry weight (Gharieb and Gadd, 2004). In
comparison with a wild type strain of S. cerevisiae, for
the GSH-deficient mutant strain, the cellular content of
Te, Co, Cu, and Mn is basically constant, and the content
of Zn and Ni increases, but that of Cr decreases, without
changing the tolerance of the cells of S. cerevisae.
However, the mutant strain displayed a high sensitivity to
selenite and cadmium, and the cellular content of both Se
and Cd increased. The experimental data indicated that
the tolerance of Te, Zn, Co, Cu, Mn, Ni and Cr is
independent of cellular glutathione activity, which
confirms the role of GSH on detoxification of Se and Cd.
The role of vacuole in the detoxification of metal ions
was investigated, and the results showed that vacuole-
deficient strain displayed much higher sensitivity and
the biosorption capacity for Zn, Mn, Co and Ni de-
creased (Ramsay and Gadd, 1997). However no sig-
nificant difference for Cd and Cu biosorption or
sensitivity to both the metal ions was observed between
wild type and mutant of S. cerevisiae. Gharieb and Gadd
(1998) found that the vacuolar-lacking strains and the
defective mutants of S. cerevisiae display higher
sensitivity to chromate and tellurite with a decrease in
the cellular content of the each metal, whereas the
tolerance to selenite increased with the cellular content
of Se. Avery and Tobin (1992) also confirmed that Sr2+
accumulation occurs mainly in the vacuole of the living
yeast cell of S. cerevisiae.
Brady et al. (1994d) proved that the copper-resistant
mechanism of S. cerevisiae was to reduce the intracel-
lular uptake of copper because the total copper content
in the copper-tolerant yeast was lower than the parent
strain when exposed to similar conditions.
The detoxification of metal ions can also be realized
by oxidation, reduction, methylation or demethylation.
Rosen (2002) proved that one mechanism of detoxifi-
cation of As(V) was the reduction of As(V) to As(III), a
process catalyzed by arsenate reductase enzymes. More
information on metal remediation by reduction mech-
anism can be referred to the review by Lovley and
Coatest (1997), including Cr6+, U6+, Te3+, Co3+, Se6+,
Pu3+ and Hg2+ reduced by various metal reducing
organisms.
Many genes involved in the uptake or detoxification
or tolerance to metal ions have been identified (Rosen,
2002). For example, the S. cerevisiae Arr4p plays an
important role in the tolerance to metal ions (As3+, As5+,
Co2+, Cr3+, Cu2+, VO43−) (Shen et al., 2003).
Based on the understanding of metal uptake
mechanism, genetic technologies, including the cell
surface display technology have been applied to
improve the performance of biomass in metal removal
from solution (Bae et al., 2003; Kuroda et al., 2002;
Wang, 2005). Kuroda et al. (2002) have constructed a
cell surface-modified yeast S. cerevisiae which dis-
plays histidine hexa-peptide, the engineered yeast that
can chelate copper ion, and possesses the property of
self-aggregation, which indicates the potential appli-
cation for bioremediation of heavy metal pollution.
9. Instrumental tools and techniques used in metal
biosorption studies
Various analytical techniques are used in metal bio-
sorption by S. cerevisiae, including atomic absorption
spectrometry (AAS), infrared absorption spectroscopy or
Fourier Transformed Infrared Spectroscopy (IR or FTIR),
scanning electron microscopy (SEM), Transmission Elec-
tron Microscopy (TEM), TEM-EDS, SEM-EDS, and X-
ray photoelectron spectrum (XPS) (Lin et al., 2005; Suh et
al., 1998a; White and Gadd, 1995).
Fourest and Volesky (1996) proved that cadmium and
carboxyl groups of the alginate formed bidentate
complex by FTIR spectra analysis. Gomes et al. (2002)
confirmed the major functional group in silver removal
was carboxyl by FTIR. Brady et al. (1994d) observed
that the copper-tolerant cell surface of S. cerevisiae
became convoluted, more spherical and smaller by SEM.
Suh et al. (1998a) confirmed the process of Pb2+ accu-
mulation in S. cerevisiae using a TEM technique. Lin et
al. (2005) utilized multi-techniques, including AAS, X-
ray powder diffraction analysis (XRD) and XPS, to
elucidate the mechanism of Au3+ removal by S. cere-
visiae. The results proved that Au3+ was reduced to Au0
by an aldehyde group of the reducing sugars as the
 J. Wang, C. Chen / Biotechnology Advances 24 (2006) 427–451 447

hyrolyzates of the polysaccharides on the peptidoglycan
layer of the yeast cells.
Techniques applied for investigation of metal bio-
sorption by other organisms can also be used in the yeast
of S. cerevisiae biosorption (see Table 6). Different
techniques provide different information on metal
biosorption, which are complementary to each other. It
is necessary to combine different techniques to explore
the mechanisms of metal biosorption.
10. Discussions and future directions
S. cerevisiae, a promising biomaterial for metal re-
moval because of its unique characteristics, has received
increasing attention during the past decades. The exis-
ting problems and progress in metal biosorption tech-
nology naturally fit for the organism of S. cerevisiae.
Therefore, the future direction in biosorption is dis-
cussed here but not limited to S. cerevisiae.
Biosorption is basically at lab scale in spite of its
development for tens of years. The mechanism was not
fully understood, and the shortcomings of biosorption
technology are two major factors that limit biosorption
application. In future work several aspects may be
considered.
studied with great efforts by utilizing various
techniques and the combination of them (Kra-
tochvil and Volesky, 1998). Important influencing
factors, such as pH and selectivity of co-ions for
biosorption systems, have not been fully
understood.
(2) Mathematical models of equilibrium and kinetics,
in particular mechanical models such as the sur-
face complexation models, to simulate multi-
metal ions or co-ions biosorption system, are
important aspects in future biosorption studies
(Liu et al., 2002a,b; Wang, 2002a) and should be
developed.
(3) Development of dynamic models to simulate the
biosorption process and offer useful information
for its application should receive more attention
(Tsezos, 2001).
(4) Molecular biotechnology, a powerful tool to
elucidate the mechanism at molecular level should
be considered more in the future to construct an
engineered organism with higher sorption capac-
ity and specificity for target metal ions (Wang and
Yang, 1996).
10.2. Application of biosorption technology

10.1. Mechanism research In spite of the failure in attempt to commercialize the

biosorption in wastewater treatment, it is necessary to
(1) The mechanisms involved in biosorption or continue to explore the various aspects relevant to the
metal–microbe interactions should be further application.

Table 6
Some techniques applied in metal biosorption studies
Main techniques References
Differential pulse polarography, directly monitoring the biosorption of metal ions from Savvaidis et al. (2003)
solution by live bacteria from mixed metal solutions, e.g. Cd(II), Zn(II) and Ni(II)
Using atomic-force microscopy (AFM) in conjunction with transmission electron microscopy (TEM) and Macaskie et al. (2000)
electron-probe X-ray microanalysis, to visualize deposition of uranyl phosphate at the cell surface
X-ray diffraction (XRD) and proton induced X-ray emission (PIXE) analyses confirmed the Ni removal Bonthrone et al. (1996)
mechanism: via intercalation to preformed the crystalline HUO2PO4·4H2O, forming Ni(UO2PO4)2·7H2O
Transmission Electron Microscopy (TEM) and Energy Dispersive X-ray (EDAX) microanalysis, to obtain information Tsezos et al. (1997)
on the localization of the metals palladium, silver, yttrium, and nickel in the different parts of the bacteria cells
Potentiometric titrations and time-resolved laser-induced fluorescence spectroscopy were used to determine Texier et al. (2000)
the binding sites of the biomass Pseudomonas aeruginosa for lanthanide ions: possible metal binding groups
are diverse, and carboxyl and phosphate groups may be main groups
The electron spin resonance (ESR) spectra of Cu(II) in bacterial cells revealed Cu(II) in the cells has the Nakajima (2002)
tetragonally distorted octahedral structure with nitrogen and oxygen as ligand atoms, indicating Cu(II)
binding with amino acid residues in the cell surface proteins
The high-resolution potentiometric titration was used to study the acid properties of the biomass in Naja et al. (2005)
order to identify the functional acidic groups for sorption and to determine their affinities by considering their
partial or total ionization equilibrium reactions
Extended X-ray absorption fine structure (EXAFS) spectroscopy to study the chemical forms of mercury, Arai et al. (2004)
cadmium and selenium accumulated in mammals, and reveal the detoxification mechanisms of these metals
Atomic force microscopy (AFM) with chemically functionalized probes to investigate the local electrostatic Ahimou et al. (2002)
properties of microbial cell
448 J. Wang, C. Chen / Biotechnology Advances 24 (2006) 427–451
(1) The physicochemical conditions, for example,
pH, multi-ionic composition should be chosen to
simulate the real wastewater on the basis of
thermodynamics and reaction kinetics studies.
(2) Optimization of the parameters of biosorption
process, including reuse and recycling by study-
ing diffusion resistance and fluid dynamics on a
sorption column or chemical engineering reactors,
such as fluidized bed reactor.
(3) Immobilization of biomaterials is another key
aspect for the purpose of biosorption application.
It is important for decreasing the cost of immo-
bilization and consequently distribution, regener-
ation and reuse of biosorbents (Tsezos, 2001).
Although continuous process of immobilized cells
has been realized at lab scale, there is still a long
way to go for biosorption commercialization.
Selection of good and cheap support materials for
biomaterial immobilization, improvement of
reuse methods, and enhancement on properties
of immobilized biosorbents such as pore ratio,
mechanical intensity and chemical stability are
also important factors for application (Hu and
Wang, 2003).
10.3. Screening of biomaterials
To screen and select the most promising biomass with
sufficiently high metal-binding capacity and selectivity for
heavy metal ions are prerequisites for a full-scale
biosorption process, this is the first major challenge in the
biosorption field (Kratochvil and Volesky, 1998).
10.4. Hybrid technology
The difficulties existing for biosorption application
urge people to consider applying hybrid technologies,
which involved combining various processes to treat real
effluent. Various biotechnology-based processes, such as
biosorption, bioreduction and bioprecipitation were
suggested. Consequently, application of living cells
rather than dead cells in biosorption received more
attention again (Malik, 2004). The above-mentioned
bioprocesses along with other non-biotechnology-based
processes, for example, chemical precipitation, flotation,
electrochemical processes, membrane technology, may
also be helpful for treating wastewater in large-scale, for
simultaneous removal of organic substances and heavy
metal ions from aqueous solution. Some examples have
been reported (Brady et al., 1994e; Riordan et al., 1997;
Thomas and Macaskie, 1998). All these processes may
possibly be realized in a single reactor, hence the
corresponding novel reactors should be designed
(Tsezos, 2001).
To sum up, S. cerevisiae, a unique and promising
biomass for metal biosorption, and is expected to
contribute to elucidate the biosorption mechanism.
Future study should be based on the advancements of
biomass processes following the well-tried and docu-
mented methodologies. Duplicating or repeating work
must be avoided (Tsezos, 2001).
Acknowledgement
The authors are grateful to the precious comments and
careful correction made by anonymous reviewers. The
authors also would like to acknowledge the financial
support from the National Natural Science Foundation of
China (Grant No. 50278045) and the Basic Research Fund
of Tsinghua University (Grant No. JC2002054).
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