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Review

Microbial and plant derived biomass for removal of


heavy metals from wastewater
Sarabjeet Singh Ahluwalia, Dinesh Goyal
*
Department of Biotechnology & Environmental Sciences, Thapar Institute of Engineering & Technology, Patiala 147 004, Punjab, India
Received 12 July 2005; received in revised form 29 November 2005; accepted 2 December 2005
Available online 19 January 2006
Abstract
Discharge of heavy metals from metal processing industries is known to have adverse eects on the environment. Conventional treat-
ment technologies for removal of heavy metals from aqueous solution are not economical and generate huge quantity of toxic chemical
sludge. Biosorption of heavy metals by metabolically inactive non-living biomass of microbial or plant origin is an innovative and alter-
native technology for removal of these pollutants from aqueous solution. Due to unique chemical composition biomass sequesters metal
ions by forming metal complexes from solution and obviates the necessity to maintain special growth-supporting conditions. Biomass of
Aspergillus niger, Penicillium chrysogenum, Rhizopus nigricans, Ascophyllum nodosum, Sargassum natans, Chlorella fusca, Oscillatoria
anguistissima, Bacillus rmus and Streptomyces sp. have highest metal adsorption capacities ranging from 5 to 641 mg g
1
mainly for
Pb, Zn, Cd, Cr, Cu and Ni. Biomass generated as a by-product of fermentative processes oers great potential for adopting an econom-
ical metal-recovery system. The purpose of this paper is to review the available information on various attributes of utilization of micro-
bial and plant derived biomass and explores the possibility of exploiting them for heavy metal remediation.
2005 Elsevier Ltd. All rights reserved.
Keywords: Biosorption; Non-living microbial biomass; Wastewater; Heavy metals
1. Introduction
Increased use of metals and chemicals in process indus-
tries has resulted in generation of large quantities of eu-
ent that contain high level of toxic heavy metals and
their presence poses environmentaldisposal problems
due to their non-degradable and persistence nature. In
addition mining, mineral processing and extractive-metal-
lurgical operations also generate toxic liquid wastes. Envi-
ronmental engineers and scientists are faced with the
challenging task to develop appropriate low cost tech-
nologies for euent treatment. Conventional methods for
removing metals from aqueous solutions include chem-
ical precipitation, chemical oxidation or reduction, ion
exchange, ltration, electrochemical treatment, reverse
osmosis, membrane technologies and evaporation recov-
ery. These processes may be ineective or extremely expen-
sive especially when the metals in solution are in the range
of 1100 mg l
1
(Nourbakhsh et al., 1994). Another major
disadvantage with conventional treatment technologies is
the production of toxic chemical sludge and its disposal/
treatment becomes a costly aair and is not eco-friendly.
Therefore, removal of toxic heavy metals to an environ-
mentally safe level in a cost eective and environment
friendly manner assumes great importance.
In light of the above, biological materials have emerged
as an economic and eco-friendly option. Biomaterials of
microbial and plant origin interact eectively with heavy
metals. Metabolically inactive dead biomass due to their
unique chemical composition sequesters metal ions and
metal complexes from solution, which obviates the neces-
sity to maintain special growth-supporting conditions.
Metal-sorption by various types of biomaterials can nd
enormous applications for removing metals from solution
and their recovery.
0960-8524/$ - see front matter 2005 Elsevier Ltd. All rights reserved.
doi:10.1016/j.biortech.2005.12.006
*
Corresponding author.
E-mail address: d_goyal_2000@yahoo.com (D. Goyal).
Bioresource Technology 98 (2007) 22432257
Rather than searching thousands of microbial species
for particular metal sequestering features, it is benecial
to look for biomasses that are readily available in large
quantities to support potential demand. While choosing
biomaterial for metal sorption, its origin is a major factor
to be taken into account, which can come from (a) micro-
organisms as a by-product of fermentation industry, (b)
organisms naturally available in large quantities in nature
and (c) organisms cultivated or propagated for biosorption
purposes using inexpensive media. Dierent non-living bio-
mass types have been used to adsorb heavy metal ions from
the environment (de Rome and Gadd, 1991; Tiemann
et al., 1999). Seaweed, mold, bacteria, crab shells and yeast
are among the dierent kinds of biomass, which have been
tested for metal biosorption or removal (Volesky and
Holan, 1995).
2. Metals in wastewater and their toxicity
Euents from textile, leather, tannery, electroplating,
galvanizing, pigment and dyes, metallurgical and paint
industries and other metal processing and rening opera-
tions at small and large-scale sector contains considerable
amounts of toxic metal ions. These toxic metals ions are
not only potential human health hazards but also to
another life forms. Toxic metal ions cause physical dis-
comfort and sometimes life-threatening illness including
irreversible damage to vital body system (Malik, 2004).
From the eco-toxicological point of view, the most danger-
ous metals are mercury, lead, cadmium and chromium(VI).
In many instances the eect of heavy metals on human is
not well understood. Metal ions in the environment bio-
accumulate and are biomagnied along the food chain.
Therefore, their toxic eect is more pronounced in animals
at higher trophic levels. Mine tailing and euents from
non-ferrous metals industry are the major sources of heavy
metals in the environment (Moore and Ramamoorthy,
1984). Among commonly used heavy metals, Cr(III), Cu,
Zn, Ni and V are comparatively less toxic then Fe and
Al. Cu is mainly employed in electric goods industry and
brass production. Major applications for Zn are galvaniza-
tion and production of alloys (Volesky and Schiewer,
2000). Cadmium has a half-life of 1030 years (Moore
and Ramamoorthy, 1984) and its accumulation in human
body aects kidney, bone and also causes cancer and its
use is increasing in industrial applications such as electro-
plating and making pigments and batteries. Chromium
compounds are nephrotoxic and carcinogenic in nature
(Chen and Hao, 1998). As a result of increasing awareness
about the toxicity of Hg and Pb, their large-scale use by
various industries has been either curtailed or eliminated
(Volesky and Schiewer, 2000). An euent treatment facil-
ity within the industry discharging heavy metals contami-
nated euent will be more ecient than treating large
volumes of mixed wastewater in a general sewage treatment
plant. Thus it is benecial to devise separate treatment pro-
cedures for scavenging heavy metals from the industrial
wastewater. Current research activity in this eld is focused
on evaluating, whether biosorption may eventually provide
such an eective and economic alternative treatment pro-
cess, while biological treatment is reasonably eective in
removing organic pollutants, heavy metals however tend
to accumulate in biological sludge, which is unt as fertil-
izers and require incineration for its disposal (Tien and
Huang, 1991).
3. Conventional methods for heavy metal removal
from industrial euents
To mitigate the heavy metal pollution, many processes
like adsorption, precipitation, coagulation, ion exchange,
cementation, electro-dialysis, electro-winning, electro-coagu-
lation and reverse osmosis have been developed.
1. Precipitation is the most common method for removing
toxic heavy metals up to parts per million (ppm) levels
from water. Since some metal salts are insoluble in water
and which get precipitated when correct anion is added.
Although the process is cost eective its eciency is
aected by low pH and the presence of other salts (ions).
The process requires addition of other chemicals, which
nally leads to the generation of a high water content
sludge, the disposal of which is cost intensive (Gray,
1999). Precipitation with lime, bisulphide or ion
exchange lacks the specicity and is ineective in
removal of the metal ions at low concentration.
2. Ion exchange is another method used successfully in the
industry for the removal of heavy metals from euents.
Though it is relatively expensive as compared to the
other methods, it has the ability to achieve ppb levels
of clean up while handling a relatively large volume.
An ion exchanger is a solid capable of exchanging either
cations or anions from the surrounding materials. Com-
monly used matrices for ion exchange are synthetic
organic ion exchange resins. The disadvantage of this
method is that it cannot handle concentrated metal solu-
tion as the matrix gets easily fouled by organics and other
solids in the wastewater. Moreover ion exchange is non-
selective and is highly sensitive to pH of the solution.
3. Electro-winning is widely used in the mining and metal-
lurgical industrial operations for heap leaching and acid
mine drainage. It is also used in the metal transforma-
tion and electronics and electrical industries for removal
and recovery of metals. Metals like Ag, Au, Cd, Co, Cr,
Ni, Pb, Sn and Zn present in the euents can be recov-
ered by electro-deposition using insoluble anodes (Gray,
1999).
4. Electro-coagulation is an electrochemical approach,
which uses an electrical current to remove metals from
solution. Electro-coagulation system is also eective in
removing suspended solids, dissolved metals, tannins
and dyes. The contaminants presents in wastewater are
maintained in solution by electrical charges. When these
ions and other charged particles are neutralized with
2244 S.S. Ahluwalia, D. Goyal / Bioresource Technology 98 (2007) 22432257
ions of opposite electrical charges provided by electro-
coagulation system, they become destabilized and pre-
cipitate in a stable form.
5. Cementation is a type of another precipitation method
implying an electrochemical mechanism in which a
metal having a higher oxidation potential passes into
solution e.g. oxidation of metallic iron, Fe(0) to ferrous
iron(II) to replace a metal having a lower oxidation
potential. Copper is most frequently separated by
cementation along with noble metals such as Ag, Au
and Pb as well as As, Cd, Ga, Pb, Sb and Sn can be
recovered in this manner.
6. Reverse osmosis and electro-dialysis involves the use of
semi-permeable membranes for the recovery of metal
ions from dilute wastewater. In electro-dialysis, selective
membranes (alternation of cation and anion mem-
branes) are tted between the electrodes in electrolytic
cells, and under continuous electrical current the associ-
ated ion migrates, allowing the recovery of metals.
4. Biosorption
Biosorption is a property of certain types of inactive,
non-living microbial biomass to bind and concentrate
heavy metals from even very dilute aqueous solution.
Biomass exhibits this property, acting just as chemical
substance, as an ion exchanger of biological origin. It is
particularly the cell wall structure of certain algae, fungi
and bacteria, which was found responsible for this phe-
nomenon (Volesky, 1990). Till now, research in the area
of biosorption suggests it an ideal alternative for decon-
tamination of metal containing euents. Advantages and
disadvantages of biosorption by non-living biomass are
as follows (Modak and Natarajan, 1995).
4.1. Advantages
Growth-independent, non-living biomass is not subject
to toxicity limitation of cells. No requirement of costly
nutrients required for the growth of cells in feed solu-
tions. Therefore, the problems of disposal of surplus
nutrients or metabolic products are not present.
Biomass can be procured from the existing fermentation
industries, which is essentially a waste after ferment-
ation.
The process is not governed by the physiological con-
straint of living microbial cells.
Because of non-living biomass behave as an ion exchan-
ger; the process is very rapid and takes place between
few minutes to few hours. Metal loading on biomass is
often very high, leading to very ecient metal uptake.
Because cells are non-living, processing conditions are
not restricted to those conducive for the growth of cells.
In other words, a wider range of operating conditions
such as pH, temperature and metal concentration is pos-
sible. No aseptic conditions are required for this process.
Metal can be desorbed readily and then recovered if the
value and amount of metal recovered are signicant and
if the biomass is plentiful, metal-loaded biomass can be
incinerated, thereby eliminating further treatment.
4.2. Disadvantages
Early saturation can be problem i.e. when metal interac-
tive sites are occupied, metal desorption is necessary
prior to further use, irrespective of the metal value.
The potential for biological process improvement (e.g.
through genetic engineering of cells) is limited because
cells are not metabolizing. Because production of the
adsorptive agent occurs during pre-growth, there is no
biological control over characteristic of biosorbent. This
will be particularly true if waste biomass from a fermen-
tation unit is being utilized.
There is no potential for biologically altering the metal
valency state. For example less soluble forms or even
for degradation of organometallic complexes.
Biosorption is a rapid phenomenon of passive metal uptake
sequestration by non-growing biomass (Beveridge and
Doyle, 1989). Results are convincing and binding capacity
of certain biomass is comparable with the commercial
synthetic cation exchange resins (Wase and Foster, 1997).
Volesky and Holan (1995) have reviewed the exhaustive list
of microbes and their metal binding capacities. Further,
sorption capacity is evaluated by sorption isotherms
described by Langmuir and Freundlich models. The uptake
of metal by two biosorbents must be compared at the
same equilibrium concentration. The adsorption is easy
to understand when it refers to a single metal situation;
however in a multi-ion situation, which is generally
encountered in euent, the assessment of sorption becomes
complicated. Most of the work exists with single metal
solution and realistic approach would be inferring results
in mixed metal solution at extreme pH and variable metal
concentration. Biosorption eciency depends upon many
factors, including the capacity, anity and specicity of
the biosorbents and their physical and chemical conditions
in euents.
5. Mechanism of metal uptake
The understanding of the mechanism by which microor-
ganisms accumulate metals is crucial to the development of
microbial processes for concentration, removal and recov-
ery of metals from aqueous solution. Metabolism-indepen-
dent metal binding to the cell walls and external surfaces is
the only mechanism present in the case of non-living bio-
mass. Metabolism-independent uptake essentially involves
adsorption process such as ionic, chemical and physical
adsorption. A variety of ligands located on the fungal walls
are known to be involved in metal chelation. These include
carboxyl, amine, hydroxyl, phosphate and sulfhydryl
S.S. Ahluwalia, D. Goyal / Bioresource Technology 98 (2007) 22432257 2245
groups. Metal ions could be adsorbed by complexing with
negatively charged reaction sites on the cell surface (Beve-
ridge and Murray, 1980; Gupta et al., 2000). The relative
importance of each functional group is often dicult to
resolve (Strandberg et al., 1981). Microbial cell wall is rich
in polysaccharide and glycoproteins such as glucans, chitin,
mannans and phospho-mannans. These polymers form
abundant source of metal binding ligands. Cell walls of
fungi present a multi-laminate architecture where up to
90% of their dry mass consists of amino or non-amino
polysaccharides (Farkas, 1980). In general, the fungal cell
wall can be regarded as a two-phase system consisting of
the chitin skeleton framework embedded in an amorphous
polysaccharide matrix (Farkas, 1980). Up to 30% of Asper-
gillus niger biomass is comprised of an association of chitin
and glucan (Muzzarelli and Tanfari, 1982). Chitin and
chitosan components of the cell wall are suggested to be
important for metal uptake (Tsezos and Volesky, 1981;
Fourest and Roux, 1992).
Muraleedharan and Venkobachar (1990) reported cop-
per binding wood-rotting fungi Ganoderma lucidum was
reduced only marginally when the biosorbent was devoid
of chitin, indicating that chitin does not play a major role
in the system. Electron paramagnetic-resonance spectra
of biosorbent indicated the presence of a free radical
(unidentied) in the cell wall matrix, which interacts with
metal, resulting in metal binding by the cells. Mechanism
of uranium binding Rhizopus arrhizus involves the amine
nitrogen of chitin crystallites and takes place in a sequence
of events (Tsezos and Volesky, 1982). Other studies in fungi
have implicated the phosphate and carboxylate groups
of the cell wall in primary binding (Akthar et al., 1996;
Kapoor and Viraraghavan, 1997).
Determination of the exact mechanism is further com-
plicated by complex solution chemistry of the metals and
the inability to determine the precise metal complex present
in the solution (Tobin et al., 1984), which is not readily
amenable to instrumental analysis (Kuyucak and Volesky,
1989). Dierences in anities between elements and their
ionic species may exist for various ligands encountered in
biological system. Cell wall of dierent fungi can vary con-
siderably in their overall composition, which leads to vary-
ing adsorption capacity. However, localization of metal(s)
has been carried out using electron microscopic and X-
ray energy dispersive analyses. X-ray photoelectron spec-
troscopy for chemical analysis is a relatively new technique
for determination of binding energy of electrons in atoms/
molecules, which depends upon the distribution of valence
charges and thus gives information about the oxidation
state of an atom/ion (Gupta et al., 2000). Electron micro-
scopic observation carried out by Mullen et al. (1989)
revealed the presence of Ag
2+
as discrete particles at or
near the cell wall of both gram-positive and gram-negative
bacteria. The presence of silver was also conrmed by
energy dispersive X-ray analysis (EDAX). Figueira et al.
(1999) using the X-ray photoelectron spectroscopy
observed that iron was present in two oxidation states,
when the brown seaweed Sargassum uitans was exposed
to Fe
2+
, while only Fe
3+
was present when biomass was
exposed to ferric ions. Further, FTIR analysis has con-
rmed that both carboxyl groups were involved in the
uptake of Fe
2+
and Fe
3+
and sulfonate groups were
responsible for Fe
3+
uptakes.
6. Removal of metal by non-living biomass of microbial and
plant origin
Removal of heavy metals from aqueous solution by
using inactive and dead biomass is an innovative and alter-
native technology for removing these pollutants. Non-
living biomass of algae, aquatic ferns and seaweeds, waste
biomass originated from plants and mycelial wastes
(Tables 1a1c and 1d) from fermentation industries are
potential biosorbents for removal of heavy metals from
aqueous solution and wastewater (Puranik and Paknikar,
1999; Ahluwalia and Goyal, 2005). Considerable potential
exists for these naturally existing, abundant and cheap
sources of biomass, for use as adsorbents. Their eciency
depends on the capacity, anity and specicity including
physico-chemical nature. Biomass related metal removal
processes may not necessarily replace existing treatment
processes but may complement them.
6.1. Removal of metals by fungal biomass
Copper biosorption by non-living wood rotting fungus
Ganoderma lucidum was studied and it was found that pro-
tein interaction with metals did not play a signicant role
in copper(II) uptake (Muraleedharan and Venkobachar,
1990). Waste mycelia of A. niger, Phanerochaete chryso-
genum and Claviceps paspali from industrial fermentation
plants were used as biosorbent for Zn from aqueous envi-
ronments in batch as well as column modes. Under opti-
mized conditions, A. niger and C. paspali were superior
to P. chrysogenum (Luef et al., 1991). Biosorption of lead
by P. chrysogenum biomass was strongly aected by pH.
At pH 45, the saturated uptake capacity for lead sorption
was higher than that of activated charcoal and that
reported for some other organisms (Niu et al., 1993). Dead
cells of Saccharomyces cerevisiae removed 40% more ura-
nium or zinc than live cultures and biosorption rapidly
reached 60% of the nal uptake value within 15 min of con-
tact and uranium was deposited as ne needle-like crystals
inside the cells and on the outer cell surface (Volesky and
May-Phillips, 1995).
Non-living waste biomass of A. niger attached to wheat
bran was used as a biosorbent for removal of Zn and Cu
from aqueous solution and metal uptake was found to be
a function of the initial metal concentration, biomass load-
ing and pH. Metal uptake decreased in the presence of co-
ions, which was dependent on the concentration of metal
ions in two compounds in aqueous solution (Modak
et al., 1996). Alkali treated biomass of A. niger referred
to as Biosorb, was found to sequester Cd
2+
, Cu
2+
, Zn
2+
,
2246 S.S. Ahluwalia, D. Goyal / Bioresource Technology 98 (2007) 22432257
Table 1a
Fungal biomass for removal of heavy metals from aqueous solution
Biosorbent Metals Adsorption
capacity (mg g
1
)
References
Aspergillus foetidus Cr(VI) 2 Prasanjit and Sumathi (2005)
Aspergillus niger Cu 5 Townsley and Ross (1986)
Modak et al. (1996)
Co 95 Kuyucak and Volesky (1989)
Au 200 Kuyucak and Volesky (1989)
Co 2.4 Sakaguchi and Nakajima (1991)
Cr, Fe Goyal et al. (2003)
Pb 30 Kim et al. (1995)
Th 22 Tsezos and Volesky (1981)
162 Gadd (1988)
U 29 Sakaguchi and Nakajima (1991)
27 Kuyucak and Volesky (1989)
Zn Luef et al. (1991),Modak et al. (1996),
Muter et al. (2002)
Aspergillus terreus Th, U 60, 10 Tsezos and Volesky (1981)
Aureobasidium pullulans Cu 6 Gadd and Mowll (1995)
Pb 56.9 Suh et al. (1998)
Ahluwalia and Goyal (2003)
Cladosporium resinae Cu 18 Gadd (1988)
de Rome and Gadd (1987)
Pb Ahluwalia and Goyal (2003)
Candida utilis Cr, Cu, Pb Muter et al. (2002)
Ganoderma lucidum Cu 24 Muraleedharan and Venkobachar (1990)
Mucor meihi Cr Tobin and Roux (1998)
Mucor rouxii Pb, Zn, Cd, Ni 17, 4.89, 6.94, 5.24 Yan and Viraraghavan (2003)
Pb 769 Lo et al. (1999)
Penicillium chrysogenum Cd, Cu, Pb 11, 9, 116 Niu et al. (1993)
Cd 56 Holan and Volesky (1995)
39 Fourest et al. (1994)
Th 142 Tsezos and Volesky (1981)
Gadd and White (1992)
U 70 Tsezos and Volesky (1981)
Zn 6.5 Niu et al. (1993)
Luef et al. (1991)
Cd, Zn, Cu, Pb Skowron ski et al. (2001)
Pleurotus sapidus Cd, Hg 127, 287 Yalcinkaya et al. (2002)
Streptoverticillium cinnamoneum Pb, Zn 57.7, 21.3 Puranik and Paknikar (1997)
Penicillium italicum Cu de Rome and Gadd (1987)
Th Gadd and White (1989)
Penicillium spinulosum Cu, Zn 0.42, 0.2 Townsley and Ross (1985)
Phanerochaete chrysosporium Cd 84.5 Gabriel et al. (1996)
Pb 2 Say et al. (2001)
Cu Yetis et al. (2000)
Rhodotorula glutinis Pb 73.5 Cho and Kim (2003)
Rhizopus nigricans Cr, Pb 47 Bai and Abraham (2001)
Zn 14 Zhang et al. (1998)
Cd, Ni, Pb 19, 5, 166 Fourest and Roux (1992)
Holan and Volesky (1995)
R. oligosporus Cr 126 Ari et al. (1999)
Cd 17.09 Aloysius et al. (1999)
R. arrhizus Ni, Cd, Zn, Pb, Cu 18, 27, 14, 56, 9.5 Fourest and Roux (1992)
Cd 30 Holan and Volesky (1995)
Cr 11 Bai and Abraham (1998)
36 Nourbakhsh et al. (1994)
Co 2.9 Sakaguchi and Nakajima (1991)
Niyogi et al. (1998)
Sag and Kutsal (1998)
Prakasham et al. (1999)
Cu 10 Gadd (1988)
de Rome and Gadd (1987),
Sag and Kutsal (1998),
Zhou and Ki (1991)
(continued on next page)
S.S. Ahluwalia, D. Goyal / Bioresource Technology 98 (2007) 22432257 2247
Ni
2+
and Co
2+
eciently up to 10% of its weight (w/w)
and exhibited higher metal binding capacity as compared
to Neurospora, Fusarium and Penicillium (Akthar et al.,
1996). The kinetics of metal binding by Biosorb indicated
that it is a rapid process and about 7080% of the
metal is removed from solution in 5 min followed by slower
rate.
Dry cells of R. arrhizus were used for removal of Fe(II),
Pb(II) and Cd(II) ions from industrial wastewater (Ozer
et al., 1997). Higher adsorption rate and adsorption capa-
city were obtained at initial metal concentration up to
100 mg l
1
in a batch reactor. Cd(II) biosorption to non-
living biomass of fungus R. arrhizus and green alga Schi-
zomeris leiblenii showed that maximum adsorption rate of
Cd(II) ions to microbial biomass was at 30 C and pH
5.0, which increased with increasing Cd(II) concentration
up to 100150 mg l
1
. Adsorption by R. arrhizus was much
higher than S. leiblenii (Ozer et al., 1997). Dried, non-liv-
ing, granulated biomass of Streptoverticillium cinnamoneum
was used for recovery of Pb and Zn at optimum pH 5.06.0
and 3.54.5, respectively. Maximum loading capacity of S.
cinnamoneum biomass pre-treated with boiling water was
57.7 mg g
1
for Pb and 21.3 mg g
1
for Zn. The metals
were eectively desorbed with dilute HCl, nitric acid and
0.1 M EDTA (Ethylene diamine tetra acetic acid).
Treatment with 0.1 M sodium carbonate permitted reuse
of desorbed biomass, which decreased loading capacity
by 1437% in subsequent cycles (Puranik and Paknikar,
1997). Waste mycelial biomass comprising of Rhizopus
nigricans from the pharmaceutical fermentation industry,
was used for adsorption of lead over a range of metal ion
concentrations, adsorption times, pH and co-ions. Uptake
process followed Langmuir and Freundlich adsorption
isotherms. Comparison of the uptake between NaOH-trea-
ted and untreated biomass showed that adsorption took
place in the chitin structure of the cell wall (Zhang et al.,
1998).
Removal of Cr(VI) from aqueous solution was carried
out in batch mode using dead biomass of fungal strains
A. niger NCIM-501, A. oryzae NCIM-637, R. arrhizus
NCIM-997 and R. nigricans NCIM-880. Basic parameters
such pH (2.08.0), initial metal ion concentration (100
500 mg l
1
), contact time (224 h) and varying biomass
concentration (0.53.0 g) were optimized. R. nigricans
and R. arrhizus possessed good specic uptake of
11 mg Cr(VI) g
1
of biomass at the pH range of 2.07.0.
Metal uptake capacity was in the order of Rhizopus nigri-
cans > Rhizopus arrhizus > Aspergillus oryzae > Aspergillus
niger (Bai and Abraham, 1998). Mucor meihi, a fermenta-
tion industry waste was found to be an eective biosorbent
for the removal of hexavalent chromium from leather
industry euent. In comparative studies with ion-exchange
resins, Mucor biomass showed biosorption levels corre-
sponding to the strongly acidic commercial resins.
Response to pH was similar to the weakly acidic resins in
solution and chromium elution characteristics were similar
to both weakly and strongly acidic resins (Tobin and Roux,
1998). Non-living free and immobilized biomass of R.
arrhizus were used to study biosorption of chromium(VI).
The removal rates were slightly more in free biomass con-
ditions over immobilized state. Stirred tank reactor studies
indicated maximum chromium biosorption at 100 rpm and
1:10 biomassliquid ratio. The uidized bed reactor was
more ecient in chromium removal than the stirred tank
reactor (Prakasham et al., 1999).
The inuences of co-cations (cadmium, copper, cobalt
and nickel) on lead and zinc biosorption by S. cinnamo-
neum and Penicillium chrysogenum in binary and multi-
metal systems was studied (Puranik and Paknikar, 1999).
Metal sorption capacity of S. cinnamoneum was observed
Table 1a (continued)
Biosorbent Metals Adsorption
capacity (mg g
1
)
References
Th 185 Tsezos and Volesky (1981)
97 Gadd et al. (1988)
Gadd and White (1992)
U 220 Tsezos and Volesky (1981)
Tsezos et al. (1989)
Tsezos and Deutschmann (1990)
Saccharomyces cerevisiae Cd Volesky et al. (1993)
Cr Nourbakhsh et al. (1994)
11.4 Omar et al. (1996)
Co 5.8 Sakaguchi and Nakajima (1991)
Pb Suh et al. (1998)
Th 119 Gadd (1988)
Cu 1740 Volesky and May-Phillips (1995)
U 55140
Cu 10 Mattuschka et al. (1993)
0.4 Huang et al. (1990)
Zn 1440 Volesky and May-Phillips (1995)
Trametes versicolor Cr Bayramoglu et al. (2003)
Cd 109 Gabriel et al. (1996)
2248 S.S. Ahluwalia, D. Goyal / Bioresource Technology 98 (2007) 22432257
to be higher than P. chrysogenum for all metals. The extent
of metal sorption depended on metal chemistry, anity for
binding sites and type of metal. The order of metal biosorp-
tion in a multi-metal system could be predicted well based
on Langmuir parameters evaluated in binary metal system.
The kinetics and mechanisms of lead(II) biosorption by
pulverized R. oligosporus were studied in batch mode (Ari
et al., 1999). At an initial lead concentration ranging from
50 to 200 mg l
1
, pH 5.0 and optimum biomass concen-
tration (0.5 g l
1
) the maximum lead adsorption was
750 mg g
1
with lead uptake capacity of 126 mg g
1
. Chro-
mium sorption by powdered biomass of R. nigricans was
optimum at pH 2, an agitation rate of 120 rpm, a temper-
ature of 45 C, a contact time of 30 min and a dosage of
0.5% (w/v) with a biomass particle size of 90 lm. High
adsorption was observed at lower initial concentration of
Cr ions (Bai and Abraham, 2001).
The biosorption of Cd(II), Pb(II) and Cu(II) onto the
dry fungal biomass of Phanerochaete chrysosporium was
investigated from articial wastewater in the concentration
range of 5500 mg l
1
(Say et al., 2001). Maximum bio-
sorption of dierent heavy metal ions was obtained at
pH 6.0 and biosorption equilibrium was established after
about 6 h and experimental biosorption data followed the
Langmuir adsorption model. The biosorption of heavy
metals by whole mycelia and selected components of A.
Table 1b
Algal biomass for removal of heavy metals from aqueous solution
Biosorbent Metals Adsorption capacity (mg g
1
) References
Ascophyllum nodosum Cd 215 Holan et al. (1993)
Co 156, 100 Kuyucak and Volesky (1988, 1989)
Ni, Pb 30, 270360 Holan and Volesky (1995)
Cd 30 Volesky and Prasetyo (1994)
Aphanothece halophytica Zn 133 Incharoensakdi and Kitjaharn (2002)
Chlorella vulgaris Ag Harris and Ramelow (1990)
Cd 111 Aksu (2001)
Cu 43 Aksu et al. (1992)
Cr 3.5 Nourbakhsh et al. (1994)
Cr, Cu, Ni Donmez et al. (1999)
U 3.95 Sakaguchi and Nakajima (1991)
Greene et al. (1986)
Chlorella fusca Pb 293 Wehreim and Wettern (1994)
Chlorella sorokiniana Cd Akhtar et al. (2003)
Cladophora crispate Cr 3 Nourbakhsh et al. (1994)
Caulerpa lentillifera Cu, Cd, Pb, Zn Apiratikul et al. (2004)
Dunaliella sp. Cr 58.3 Donmez and Aksu (2002)
Fucus vesiculosus Cd 73 Holan et al. (1993)
Ni, Pb 17, 220371 Holan and Volesky (1995)
Fucus spiralis Cd 64 Cordeo et al. (2004)
Ecklonia maxima Cd Stirk and Staden (2002)
Laminaria japonica Cd Yun et al. (2001)
Laurencia obtuse Cr Hamdy (2000)
Lyngbya taylorii Cd, Pb, Ni, Zn Klimmek et al. (2001)
Phormidium laminosum Cu, Ni, Zn Blanco et al. (1998)
Pilayella littoralis Al, Cd, Co, Cr, Ni, Zn Carrilho and Gilbert (2000)
Pachymeniopis sp. Cr(VI) 225 Lee et al. (2000)
Oscillatoria anguistissima Zn 641 Ahuja et al. (1999)
Zn, Cu, Co Mohapatra and Gupta (2005)
Spirogyra sp. Cr Gupta et al. (2001)
Scenedesmus quadricula Cd, Cu, Zn Harris and Ramelow (1990)
Scenedesmus obliquus Cr, Cu, Ni Donmez et al. (1999)
Scenedesmus abundans Cd, Cu Terry and Stone (2002)
Scenedesmus incrassatulus Cr, Cd, Cu Pena-Castro et al. (2004)
Sargassum uiyans Cu Schiwer and Volesky (1996)
51 Kratochvil et al. (1997)
Sargassum natans U Kuyucak and Volesky (1989)
Cd 135 Holan et al. (1993)
Ni, Pb 2444, 220270 Holan and Volesky (1995)
Sargassum sp. Zn da Cotsa et al. (2001)
Sargassum sp. Cu 38, Volesky et al. (2003),Padilha et al. (2005)
Cd, Zn, Cu 157, 118, 77 Valdman and Leite (2000)
Cd 120 Cruz et al. (2004)
Tetraselmis suecica Cd Perez-Rama et al. (2002)
Ulothrix zonata Cu Nuhoglu et al. (2002)
Ulva lactuca Hg Zeroual et al. (2003)
S.S. Ahluwalia, D. Goyal / Bioresource Technology 98 (2007) 22432257 2249
niger, R. oryzae and Mucor rouxii was studied (Baik et al.,
2002). Binding of copper, cadmium, nickel and zinc was
considerably improved by treating the cell wall fraction
with 4 M NaOH at 121 C.
Table 1c
Bacterial biomass used for metal removal
Biosorbent Metals Adsorption capacity (mg g
1
) References
Pseudomonas uorescens Th, U 15, 6 Tsezos and Volesky (1981)
Pseudomonas sp. U Pons and Fuste (1993)
Ochrobactrum anthropi Cr, Cd, Cu Ozdemir et al. (2003)
Thiobacillus ferrooxidans Zn 82 Baillet et al. (1998)
Cr Celaya et al. (2000)
Rhodococcus erythropolis Cd, Zn Plette et al. (1996)
Ocimum basilicium Cr Melo and DSouza (2004)
Sphingomonas paucimobilis Cd Tangaromsuk et al. (2002)
Bacillus rmus Pb, Cu, Zn 467, 381, 418 Salehizadeh and Shojaosadati (2003)
B. coagulans Cr(VI) 39.9 Srinath et al. (2002)
B. megaterium Cr(VI) 30.7 Srinath et al. (2002)
Zoogloea ramigera Cr(VI) 2 Nourbakhsh et al. (1994)
Cu 29 Aksu et al. (1992)
Streptomyces noursei Cd, Cu, Ni 3.4, 9, 0.8 Mattuschka and Straube (1993)
Pb, Zn, Ag, Co 36, 1.6, 38 Mattuschka and Straube (1993)
Cr, Pb 1.2 Mattuschka and Straube (1993)
1.8, 55 Mattuschka et al. (1993)
S. longwoodensis Pb, U 100, 440 Friis and Myers-Keith (1986)
S. rimosus Zn 30 Mameri et al. (1999)
Table 1d
Plant derived biomass used for metal removal
Biosorbent Metals Adsorption capacity (mg g
1
) References
Activated baggase carbon Cr Mor et al. (2002)
Alfalfa Cu, Pb 20, 43 Tiemann et al. (1999)
Artocarpus heterphyllus Cd Inbaraj and Sulochana (2004)
Azolla liculoides Pb 93 Sanyhumbi et al. (1998)
Banana pith (Musacea zingiberales) Pb, Cu, Ni, Cr, Zn Low et al. (1995)
Hydrilla verticillata casp. and Salvinia sp. Cu Elankumaran et al. (2003)
Carrot pulp Pb, Ni, Zn, Fe Joshi et al. (2003)
Ceratophyllum demersum Cu, Pb, Zn 6.17, 45, 14 Keskinkan et al. (2004)
Cupressus Female Cone Cr(VI) 119 Murugan and Subramanian (2003)
Eucalyptus wood powder Pb, Ni, Zn, Fe Joshi et al. (2003)
Eucalyptus bark Cr(VI) Sarin and Pant (2006)
Ground corncobs Cd, Cu, Pb Vaughan et al. (2001)
Ni, Zn
Hydrilla verticillata casp. Cu Elankumaran et al. (2003)
Larrea tridenta Cu 24 Gardea-Torresdey et al. (2004a,b)
Olive mill solid residue Hg, Pb, Cu Pagnanelli et al. (2002)
Zn, Cd
Paper mill sludge Pb, Cu, Ag Calace et al. (2003)
Cd
Pb, Ni, Zn, Fe Joshi et al. (2003),Ahluwalia and Goyal (2004)
Sago processing waste Cu, Pb 12.4, 46.6 Quek et al. (1998)
Sawdust Zn, Ni, Cd, Cu, Pb Marin and Ayele (2002)
Cr(VI) Acar and Malkoc (2004)
Wheat stem and Spent Babul Bark Ni Verma and Shukla (2000)
Oryza sativa L. hush Pb Zulkali et al. (2006)
Waste tea-leaves Pb, Ni, Zn, Fe Joshi et al. (2003),Ahluwalia and Goyal (2005)
Wola globosa Cd, Cr Upatham et al. (2002)
Water hyacinth roots Cr Low et al. (1997)
Helianthus annuus L. (Sunower) Cu Lin et al. (2003)
Allium sativum L. (Garlic) Cd Jiang et al. (2001)
Grape stalk waste Cu, Ni 15.9, 18.1 Villaescusa et al. (2004)
Hemidesmus indicus Pb Chandrasekhar et al. (2003)
Myriophyllum spicatum Pb, Cu, Cd Keskinkan et al. (2003)
2250 S.S. Ahluwalia, D. Goyal / Bioresource Technology 98 (2007) 22432257
6.2. Removal of metals by algal biomass
Chromium sorption by non-living biomass of Chlorella
vulgaris and Cladophora crispata along with bacteria and
fungi was studied and it was observed that pH aected
metal uptake capacity (Nourbakhsh et al., 1994). Maxi-
mum adsorption was observed at a temperature range of
2535 C, which increased with increasing metal concen-
trations up to 200 mg l
1
. The short-term accumulation
of chromate by Anabaena sp. and Synechococcus sp.
PCC6301 was rapid with relatively low level of sorption
to the cell wall. The process was not energy dependent
but was regulated bychromate concentration described by
Freundlich adsorption isotherm. On decreasing pH, chro-
mate accumulation increased in both the species. Over a
longer growth period Anabaena variabilis was capable of
reducing chromate(VI) to chromium(III) and accumulating
it. Synechococcus PCC 6301 showed no further interaction
with chromate concentration over the same period after an
initial biosorption (Garnham and Green, 1995). In brown
seaweeds, 9095% removal of Hg and Cd from industrial
wastewater was observed (Wilson and Edween, 1995).
Biosorption of Cu(II), Fe(II), Ni(II) and Zn(II) by non-
viable biomass of the cyanobacterium Phormidium lamino-
sum from single and binary metal solution was extremely
rapid and consisted of a single phase (Blanco et al.,
1998). The presence of a second metal in solution decreased
the binding of Fe(II), Ni(II) and Zn(II) due to chemical
blocking of the carboxyl groups of biomass but this had
no eect on the binding of Cu(II). Washing with dilute
acids was much faster and eective than NaOH, NaCl,
CaCl
2
and ultra pure water reaching equilibrium within
30 min and the eciency was independent of the concentra-
tion of desorbent. Biosorption of Cu(II), Ni(II) and Cr(VI)
from aqueous solution by dried algae Chlorella vulgaris,
Scenedesmus obliquus and Synechocystis sp. was tested
under laboratory conditions as a function of pH, initial
metal ion and biomass concentration (Donmez et al.,
1999). Results showed the inuence of algal biomass
concentration on metal uptake by all species and both Fre-
undlich and Langmuir adsorption models described the
short-term biosorption. Out of 48 dierent species of red,
brown and green algae examined for chromate adsorption,
excellent adsorption was observed by Pachymeniopsis sp.,
which had high selectivity for chromate among other heavy
metals ions such as cadmium and manganese. It also exhib-
ited a typical Langmuir isotherm with q
max
of 225 mg g
1
and K 1106 mg ml
1
(Lee et al., 2000). Biosorption of tri-
valent chromium by protonated brown algal biomass of
Eklonia revealed three types of functional groups. The car-
boxyl group was the chromium-binding site within the pH
range of 15 at which chromium does not precipitate. The
pK value and number of carboxyl groups were estimated to
be 4.6 0.1 and 2.2 0.1 mmol g
1
, respectively (Yun
et al., 2001). Adsorption of Cu(II) from aqueous solution
by Ulothrix zonata was studied using batch adsorption
technique (Nuhoglu et al., 2002). The equilibrium biosorp-
tion level was found to be a function of contact time at dif-
ferent initial metal ion concentration and the adsorption
data showed the Langmuir adsorption model. The free
energy change for the adsorption process was found to
be 12.60 kJ mol
1
. Dried Caulerpa lentillifera was shown
to have adsorption potential for Cu, Cd, Pb and Zn (Api-
ratikul et al., 2004). The adsorption equilibrium followed
the Freundlich isotherm. The adsorption of metals from
binary mixture of heavy metal was competitive and the
adsorption capacity of any single metal decreased by 10
40% in the presence of the other metallic species and the
overall adsorption capacity of the algae decreased by 30
50%.
Uranium biosorption by powered biomass of lake-har-
vested water bloom cyanobacterium Microcystis aeruginosa
(Li et al., 2004) was optimum at 4.08.0 pH. The batch
sorption reached the equilibrium within 1 h and followed
the Freundlich isotherm model. Uranium was eectively
desorbed with 0.01 N HCl. This naturally abundant bio-
mass otherwise nuisance cyanobacterium exhibited good
potential for application in removal of uranium from aque-
ous solution. The waste biomass of Sargassum sp. for the
recovery of ionic copper from simulated semiconductor
euents was investigated (Padilha et al., 2005) using con-
tinuous system comprising of four column reactors lled
with the biomass which showed high operation stability
and completely biosorbed the ionic copper from 63 l of
copper sulphate solution, 72 l of copper chloride solution
and 72 l of copper nitrate solution, all the solutions con-
taining 500 mg l
1
copper. Copper concentration was
found to be less than 0.5 mg l
1
in the euent from the
outlet of the reactor. Five dierent brown seaweeds, Bifur-
caria bifurcata, Saccorhiza polyschides, Ascophyllum nodo-
sum, Laminaria ochroleuca and Pelvetia caniculata were
studied for their ability to remove cadmium from aqueous
solution (Lodeiro et al., 2005). Kinetics of cadmium
adsorption by all algae was relatively fast with 90% of total
adsorption occurring in less than 1 h.
6.3. Removal of metals by bacterial biomass
Bacteria may uptake and accumulate a signicant
amount of metal ions, resulting in the transfer of metals
to a contaminated matrix of biomass (Smith et al., 1994).
When suspension of dead biomass of actinomycetes from
industrial fermentation, was mixed with wastewater, the
biosorption of cadmium cations occurred due to negatively
charged sites on bacterial cell wall (Butter et al., 1996). Zinc
biosorption capacity of Streptomyces rimosus biomass was
studied in batch mode and after heat treatment (Mameri
et al., 1999). The optimum conditions of biosorption were
a contact time of 4 h, biomass particle size 140250 lm,
ambient temperature, a stirring speed of 250 rpm and pH
7.5. The biosorption capacity was 30 mg of Zn g
1
of
biomass, which increased up to 80 mg following chemical
treatment by NaOH. The removal of chromium, cadmium
and copper from dilute aqueous solution using dead
S.S. Ahluwalia, D. Goyal / Bioresource Technology 98 (2007) 22432257 2251
polysaccharide producing Ochrobactrum anthropi, isolated
from activated sludge (Ozdemir et al., 2003). Optimum
adsorption pH values of Cr(VI), Cd(II) and Cu(II) were
2.0, 8.0 and 3.0, respectively. Experimental showed the
inuence of initial metal concentration on the metal uptake
for dried biomass. Both the Langmuir and Freundlich
adsorption were suitable for describing the short term
biosorption of Cr(VI), Cd(II) and Cu(II) by O. anthropi.
6.4. Removal of metals by plant derived biomass
The sorption capacity of dierent biosorbents like dried
mycelium of some species of fungi, baggase, rice husk and
fermented baggase was examined to remove cyanide from
industrial euent (Azab et al., 1995). Banana pith (Musacea
zingiberales) was evaluated for its ability to sorb metal ions
from electroplating waste and synthetic solutions under
both batch and continuous ow conditions. The sorption
was observed to be both pH and concentration dependent
with pH 45 being optimum. The equilibrium data followed
the Langmuir isotherm model with maximum capacity of
8.55 and 13.46 mg g
1
of Cu in electroplating waste and
synthetic solution, respectively (Low et al., 1995).
Waste biomass such as wheat stem and babul bark was
used to remove nickel from euent of an electroplating
industry (Verma and Shukla, 2000). The removal of nickel
was 210% less as compared to synthetic solution under
similar conditions. Wheat straw activated carbon could
remove 100% Ni(II) from initial nickel concentration of
25 mg l
1
at pH 4.0 in 4 h at 36 2 C. The uptake capa-
city of dierent metals by Quercus ilex phytomass (root,
stem and leaf) was found to be in the order of roots
Ni > Cd > Pb > Cu > Cr; stem Ni > Pb > Cu > Cd > Cr;
and leaf Ni > Cd > Cu > Pb > Cr (Prasad and Freitas,
2000). Desorption with 10 mM Na
4
EDTA was eective
and there existed the possibility of recycling the phytomass.
Biosorption capacity of plant biomass of Indian Sarapa-
rilla (Hemidesmus indicus) was studied with toxic heavy
metals like As, Se, Zn, Fe, Ni, Co, Pb, Mn, Hg, Cr and
Cu. Lead was preferentially removed followed by Cr and
Zn at concentration less than 250 mg l
1
and with biomass
quantity above 2 g. Presence of co-ions (As, Se and Hg) did
not eect the Pb removal but Zn and Cr uptake decreased.
Metal loaded biomass was regenerated with HNO
3
and
reused for three cycles without any loss in metal retention
capacity (Chandrasekhar et al., 2003). Adsorption of
Cd(II) and Ni(II) was found to be higher than Cr(VI)
and Zn(II) when phosphate treated rice husk was used as
an adsorbent (Ajmal et al., 2003). Sorption of Cd(II) was
dependent on contact time, concentration, temperature,
adsorbent doses and pH of the solution and follows the
Langmuir constant. Recovery of Cd(II) from aqueous solu-
tion by column operation was found to be higher than
batch process. Adsorption of Cr(VI) with activated rice
husk carbon and activated alumina (Bishnoi et al., 2004)
and with formaldehyde treated saw dust and sulphuric acid
treated saw dust carbon (Garg et al., 2004) was investi-
gated. The amount of Cr(VI) adsorbed increased with
increase in dose of these adsorbents and their contact time.
Reduction rate of Cr(VI) to Cr(III) by Ecklonia sp. bio-
mass increased with decreasing pH, which appeared in the
solution phase or was partly bound to the biomass (Park
et al., 2004). The jack fruit peel (Artocarpus heterophyllus)
was treated with sulphuric acid to produce a carbonaceous
product, which was used as an adsorbent for the removal
of Cd(II) from aqueous solution (Inbaraj and Sulochana,
2004). Kinetic analysis made with Lagergren pseudo-rst
order, Ritchie second order and modied Ritchie second
order models showed better ts with modied Ritchie
second order model. The LangmuirFreundlich (Sips
equations) model best denes the experimental equilibrium
data among the three isotherm models (Freundlich, Lang-
muir and LangmuirFreundlich) used. A complete recov-
ery of the adsorbed metals ions from the spent adsorbent
was achieved by using 0.01 M HCl.
The adsorption of activated carbon prepared from apri-
cot stones, to remove Ni(II), Co(II), Cd(II), Cu(II), Pb(II),
Cr(III) and Cr(VI) ions from the aqueous solution was
studied (Kobya et al., 2005). Highest adsorption occurred
at pH 12 for Cr(VI) and at pH 36 for the remaining
metal ions. Further, adsorption capacities for these metals
were found to in the order of Cr(VI) > Cd(II) > Co(II) >
Cr(III) > Ni(II) > Cu(II) > Pb(II), respectively. Potential
of modied lignocellulosic bre/jute was assessed for
adsorption of heavy metal ions like Cu(II), Ni(II) and
Zn(II) from aqueous solution (Shukla and Pai, 2005).
The dye loaded jute bre showed adsorption capacity of
8.4, 5.26 and 5.95 mg g
1
for Cu(II), Ni(II) and Zn(II),
respectively, whereas oxidized jute bre showed 7.73, 5.57
and 8.02 mg g
1
against 4.23, 3.37 and 3.55 mg g
1
for
unmodied jute bre which followed Langmuir adsorption
model. Desorption eciency, regenerative and reuse capa-
city of these adsorbent were also assessed for three succes-
sive adsorptiondesorption cycles. The sorption of lead by
tree fern, an agricultural by product was carried out using
agitated and baed system (Ho, 2005). The optimum pH
for lead removal was 4.9. Also pseudo-second-order kinet-
ics analysis was performed to determine the rate constant
of sorption, the equilibrium sorption capacity, and the ini-
tial sorption rate. Removal of cadmium from aqueous
solution by Moringa oleifera seed was studied in batch
sorption experiments as a function of biomass dosage,
contact time, metal concentration, particle size and pH
(Sharma et al., 2006). Fourier transform infrared spectro-
scopy indicated the amino-Cd interaction responsible for
this sorption. These nding opens up new avenues in the
removal of toxic metals by shelled M. oleifera seeds from
water bodies as low cost, domestic and environment
friendly safe technology.
7. Development of biosorbent
Biosorption is a complex process, mainly comprising of
ion exchange, chelation and adsorption by physical forces
2252 S.S. Ahluwalia, D. Goyal / Bioresource Technology 98 (2007) 22432257
and entrapment in inter and intra-brillar capillaries and
space of the structural polysaccharide network as a result
of the concentration gradient and diusion. There are sev-
eral chemical groups that would attract and sequester the
metals in biomass. Acetamide groups of chitin, structural
polysaccharides of fungi, amino and phosphate groups of
nucleic acids, amide, amine, sulfhydryl and carboxyl
groups in the proteins and hydroxyls in polysaccharide of
marine algae belonging to Phaeophyceae, Rhodophyceae
and Chlorophyceae are some of the examples. It does not
necessarily mean that presence of functional groups guar-
antees biosorption, perhaps due to steric, conformational
or other eects.
Several proprietory biosorption processes were devel-
oped and commercialized in the early 1990s using Alga-
SORB
TM
(Brierley et al., 1986) and AMT-Bioclaim
TM
(Darnall, 1991) but did not attract widespread adoption
due to a lack of understanding of the underlying mech-
anism of metal-sorption, proper selection of industrial
euents for pilot testing and scaling up of the process
(Kratochvil and Volesky, 1998). Routinely most prospec-
tive biosorbents are discovered by trial and experimenta-
tion. Many microorganisms can be used as biosorbents.
Fungi in particular have unique metal adsorption charac-
teristics and are easy to cultivate (Gadd, 1988). They can
successfully absorb toxic metal ions Zn
2+
, Cu
2+
, Cd
2+
,
Pb
2+
, Ni
2+
, Fe
2+
, Mn
2+
and Cr
6+
from aqueous solution.
But the process is susceptible to complexation or precipita-
tion of the metal, making it unavailable for binding due to
the presence of anions such as phosphate. Some biosorbent
have broad range, which bind the majority of heavy metals
with no specic activity, while others are specic for certain
metals. Biosorption studies have been carried out using
easily available biomass in their native state or after simple
processing. Among a variety of biomaterials, the majority
has been focused on bio-waste generated as a by-product
of large-scale industrial fermentation (Puranik and Pak-
nikar, 1999), olive mill solid residues (Pagnanelli et al.,
2002), activated sludge from sewage treatment plants
(Hammaini et al., 2003; Sag et al., 2003), biosolids (Norton
et al., 2003), aquatic macrophytes (Keskinkan et al., 2003)
and other plant derived materials (Gardea-Torresdey et al.,
2004a,b).
Norton et al. (2003) used dewatered activated sludge
from a sewage treatment plant for removing zinc from
aqueous solution and recorded adsorption capacity of
0.56 mM g
1
of biosolids. Use of biosolids for zinc adsorp-
tion was favorable compared to the bio-adsorption rate of
0.3 mmol g
1
by seaweed Durvillea potatorum (Aderhold
et al., 1996). Keskinkan et al. (2003) studied adsorption
characteristics of copper, zinc and lead on submerged
aquatic plant, Myriophyllum spicatum and found the
adsorption capacities to be 46.60 mg l
1
for lead,
15.59 mg g
1
for zinc and 10.37 mg g
1
for copper. Pagna-
nelli et al. (2002) carried out a preliminary study on the use
of olive mill residue as a heavy metal sorbent material.
They found that copper was maximally adsorbed in the
range of 5.013.5 mg g
1
under dierent operating condi-
tions. The biosorption capacity of copper, cadmium and
zinc on dried activated sludge was 0.32 m mol g
1
for metal
systems such as CuCd; 0.29 m mol g
1
for CuZn and
0.32 m mol g
1
for CdZn. Results showed that biomass
had a preference for copper followed by cadmium and zinc
(Hammaini et al., 2003). Another inexpensive source of
biomass is seaweeds comprising mainly the marine
macro-algae. However, most of the studies on the up-
take of toxic metals by marine algae and to some extent
by freshwater algae are focused on toxicological aspects,
metal accumulation and pollution indicators. There has
been no focus on the technological aspects of metal
removal by algal biomass.
8. Conclusion
Metabolic independent processes can mediate the bio-
logical uptake of heavy metal cations. Biosorption oers
an economically feasible technology for ecient removal
and recovery of metal(s) from aqueous solution. The pro-
cess of biosorption has many attractive features including
the selective removal of metals over a broad range of pH
and temperature, its rapid kinetics of adsorption and
desorption and low capital and operation cost. Biosorbent
can easily be produced using inexpensive growth media or
obtained as a by-product from industry. It is desirable to
develop biosorbents with a wide range of metal anities
that can remove a variety of metal cations. These will be
particularly useful for industrial euents, which carry
more than one type of metals. Alternatively a mixture of
non-living biomass consisting of more than one type of
microorganisms can be employed as biosorbents. Such
Combo biosorbents have to be tested for commercial
applications. The use of immobilized biomass rather than
native biomass has been recommended for large-scale
application but various immobilization techniques have
yet to be thoroughly investigated for ease, ecacy and cost
eectivity.
Biosorption processes are applicable to euents con-
taining low concentrations of heavy metals for an extended
period. This aspect makes it even more attractive for treat-
ment of dilute euent that originates either from an indus-
trial plant or from the primary wastewater treatment
facility. Thus biomass-based technologies need not neces-
sarily replace the conventional treatment routes but may
complement them. At present, information on dierent
biosorbent materials is inadequate to accurately dene
the parameters for process scale up and design perfection
including reliability and economic feasibility. To provide
an economically viable treatment, the appropriate choice
of biomass and proper operational conditions has to be
identied. To predict the dierence between the uptake
capacities of the biomass, the experimental results should
be tested against an adsorption model. The development
of a packed bed or uidized-bed biosorption model would
be helpful for evaluating industrial-scale biosorption
S.S. Ahluwalia, D. Goyal / Bioresource Technology 98 (2007) 22432257 2253
column performance, based on laboratory scale experi-
ments and to understand the basic mechanism involved
in order to develop better and eective biosorbent.
Critical analysis reveals that not all metal-polluted
wastewater-generating industries have the interest or the
capability to treat euents and most of the industries opt
only for basic treatment techniques simply to comply with
regulations. To attract more usage of biosorption, strate-
gies have to be formulated to centralize the facilities for
accepting the used biosorbent where processing of the bio-
sorbent can be done to either regenerate the biomass or
convert the recovered metal into usable form. This will fur-
ther require an interdisciplinary approach with integration
of metallurgical skills along with sorption and wastewater
treatment to apply biosorption technology for combating
heavy metal pollution in aqueous system.
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