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 Critical Reviews in Biotechnology

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Use of Algae for Removing Heavy Metal Ions From

Wastewater: Progress and Prospects

S. K. Mehta & J. P. Gaur

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Critical Reviews in Biotechnology, 25:113–152, 2005
Copyright
c Taylor & Francis Inc.
ISSN: 0738-8551 print / 1549-7801 online
DOI: 10.1080/07388550500248571
Use of Algae for Removing Heavy Metal
Ions From Wastewater: Progress
and Prospects
S. K. Mehta and J. P. Gaur
Laboratory of Algal Biology,
Department of Botany, Banaras
Hindu University, Varanasi,
India
Address correspondence to S. K.
Mehta, Laboratory of Algal Biology,
Department of Botany, Banaras Hindu
University, Varanasi, 221 005, India.
E-mail: skmehta12@rediffmail.com
ABSTRACT Many algae have immense capability to sorb metals, and there
is considerable potential for using them to treat wastewaters. Metal sorption
involves binding on the cell surface and to intracellular ligands. The adsorbed
metal is several times greater than intracellular metal. Carboxyl group is most im-
portant for metal binding. Concentration of metal and biomass in solution, pH,
temperature, cations, anions and metabolic stage of the organism affect metal
sorption. Algae can effectively remove metals from multi-metal solutions. Dead
cells sorb more metal than live cells. Various pretreatments enhance metal sorp-
tion capacity of algae. CaCl2 pretreatment is the most suitable and economic
method for activation of algal biomass. Algal periphyton has great potential
for removing metals from wastewaters. An immobilized or granulated biomass-
filled column can be used for several sorption/desorption cycles with unaltered
or slightly decreased metal removal. Langmuir and Freundlich models, com-
monly used for fitting sorption data, cannot precisely describe metal sorption
since they ignore the effect of pH, biomass concentration, etc. For commercial
application of algal technology for metal removal from wastewaters, emphasis
should be given to: (i) selection of strains with high metal sorption capacity,
(ii) adequate understanding of sorption mechanisms, (iii) development of low-
cost methods for cell immobilization, (iv) development of better models for
predicting metal sorption, (v) genetic manipulation of algae for increased
number of surface groups or over expression of metal binding proteins, and
(vi) economic feasibility.
KEYWORDS algae, biosorption, heavy metal, immobilization, isotherm, ion exchange,
seaweed, periphyton
I. INTRODUCTION
Heavy metal pollution of waterbodies due to indiscriminate disposal of in-
dustrial and domestic wastes threatens all kinds of inhabiting organisms (De
Filippis and Pallaghy, 1994). Therefore, it is necessary to alleviate heavy metal
burden of wastewaters before discharging them into waterways. A number of
physicochemical methods, such as chemical precipitation, adsorption, solvent
extraction, ion exchange, membrane separation, etc., have been commonly
113
employed for stripping toxic metals from wastewaters
(Eccles, 1999). However, these methods have several dis-
advantages, such as incomplete metal removal, expen-
sive equipment and monitoring system requirements,
high reagent or energy requirements and generation of
toxic sludge or other waste products that require dis-
posal. Further, they may be ineffective or extremely ex-
pensive when metal concentration in wastewater is in
the range 10–100 mg l−1 .
The use of biological processes for the treatment of
metal enriched wastewaters can overcome some of the
limitations of physical and chemical treatments and
provide a means for cost-effective removal of metals.
A great deal of interest has recently been generated
using different kinds of inexpensive biomass for ad-
sorbing and removing heavy metals from wastewater
(Volesky and Holan, 1995). In this context, accumu-
lation of metals by microorganisms, including algae,
has been known for a few decades, but has received in-
creased attention only in recent years because of its po-
tential for application in environmental protection or
recovery of precious or strategic metals (Tsezos, 1985,
1986; Volesky, 1987; Malik, 2004). Metal accumulation
capacity of algal biomass is either comparable or some-
times higher than chemical sorbents (see Tables 1 and
2). Therefore, algal biomass may serve as an econom-
ically feasible and efficient alternative to the existing
physicochemical methods of metal removal and recov-
ery from wastewaters. The term “biosorption” has been
more frequently used instead of accumulation; how-
ever, it should be used for the adsorption of metal
ions on the dead biomass, and it includes metal ion
binding on extracellular as well as intracellular ligands
(Volesky and Holan, 1995; Aksu, 1998). However, the
term “biosorption” has been rather loosely used for de-
scribing metal accumulation by live biomass as well.
This review explores the prospects of using algae for
removing heavy metal ions from wastewaters. After a
brief account of accumulation of metals by algae and
the mechanisms involved, the paper discusses at length
metal removal by algae and possibilities of using this
technology on a commercial scale.
II. ACCUMULATION OF METALS
BY ALGAE
Algae accumulate high concentrations of metals de-
pending on their concentration in the external environ-
ment. The concentration factor for heavy metals varies
S. K. Mehta and J. P. Gaur
greatly in different algal species, but it increases as the
metal concentration in the water decreases (Kelly, 1988;
Sharma and Azeez, 1988). Since the amount of metal
accumulated by algae is related with the concentration
of metal in water, it may be possible to use metal content
of indigenous algae for biomonitoring of metal pollu-
tion in a waterbody (De Filippis and Pallaghy, 1994).
Furthermore, the inherent metal accumulation capabil-
ity of algae could be used to alleviate the burden of
toxic metal load, and to recover precious metals (e.g.,
gold and silver) from wastewaters.
The accumulation of heavy metals in algae involves
two processes: an initial rapid (passive) uptake followed
by a much slower (active) uptake (Bates et al., 1982).
During the passive uptake, metal ions adsorb onto the
cell surface within a relatively short span of time (few
seconds or minutes), and the process is metabolism-
independent. Active uptake is metabolism-dependent,
causing the transport of metal ions across the cell mem-
brane into the cytoplasm. In some instances, the trans-
port of metal ions may also occur through passive dif-
fusion owing to metal-induced increase in permeability
of the cell membrane (Gadd, 1988).
Bates et al. (1982) described a simple method for dis-
tinguishing adsorbed from intracellular metal in algal
cells. The method involves washing cells with 2 mM
EDTA, which removes surface-bound metal, leaving be-
hind the intracellular fractions. The intracellular metal
can be quantified after digesting the EDTA-washed
cells. The surface adsorbed metal can be quantified by
subtracting intracellular metal from the total metal ac-
cumulated by the cell. The method has been commonly
used by several researchers to distinguish adsorbed from
intracellular metal in spite of inevitable removal of a
small fraction of intracellular metal during EDTA wash-
ing (Bates et al., 1982). The relative importance of metal
adsorption and uptake may vary with algal species and
metal ions (Trevors, Stratton, and Gadd, 1986). Gen-
erally, adsorption contributes much more, even >80%
(Mehta, Singh, and Gaur, 2002; Mehta, Tripathi, and
Gaur, 2000), than uptake to total metal accumulation
by algal cells. However, there are a few reports sug-
gesting a greater contribution of uptake than adsorp-
tion or almost similar contribution of both the pro-
cesses to total metal accumulation (Avery, Codd, and
Gadd, 1998). After the first minute of exposure to Cu,
>90% of total metal content was found adsorbed on
the surface of Scenedesmus subspicatus (Knauer, Behra,
and Sigg, 1997). With the passage of time, the relative
114
TABLE 1 Recent reports on metal sorption capacity of some algae.
a
Sorption

Metal Alga (mmol g−1 ) (mg g−1 ) Reference

Al Laminaria japonica 2.79 75.28 Lee et al., 2004

Pilayella littoralis 2.00 53.96 Carrilho and Gilbert, 2000
Spirulina sp. 0.003 0.08 Chojnacka, Chojnacki, and Górecka, 2004
Au Chlorella vulgaris 0.13 25.02 Ting, Teo, and Soh, 1995
Sargassum fluitans 0.005 1.00 Niu and Volesky, 2000
Co Oscillatoria angustissima 0.26 15.32 Mohapatra and Gupta, 2005
Pilayella littoralis 0.56 55.40 Carrilho and Gilbert, 2000
Spirulina sp. 0.0002 0.01 Chojnacka, Chojnacki, and Górecka, 2004
Ulva reticulata 0.78 45.97 Vijayaraghavan et al., 2005
Cd Anabaena nodosum 0.087 9.78 Sandau et al., 1996
A. nodosum 0.81 91.04 Chong and Volesky, 1996
Ceramium rubrum 0.07 8.21 Sandau et al., 1996
Chlorella vulgaris 0.11 12.48 Sandau, Sandau and Pulz, 1996
Costera marina 0.09 9.55 Sandau et al., 1996
Ecklonia maxima 0.74 83.50 Feng and Aldrich, 2004
Fucus vesiculosus 0.09 9.78 Sandau et al., 1996
Gigartina salicornia 0.16 17.98 Hashim and Chu, 2004
G. acicularis 0.09 9.67 Sandau et al., 1996
Gracilaria fisheri 0.63 70.81 Chaisuksant, 2003
Laminaria digitata 0.09 9.78 Sandau et al., 1996
L. japonica 1.30 146.12 Yin et al., 2001
L. japonica 1.21 125.89 Lee et al., 2004
L. saccharina 0.08 9.34 Sandau et al., 1996
Lyngbya taylorii 0.37 41.59 Klimmek et al., 2001
Nannochloropsis oculata 0.89 100.04 Zhou, Huang, and Lin, 1998
Padina pavonia 1.10 123.64 Ofer, Yerachmiel, and Shmuel, 2003
Padina sp. 0.75 84.30 Sheng et al., 2004b
Phaeodactylon tricornutum 0.58 64.97 Zhou, Huang, and Lin, 1998
Pilayella littoralis 0.43 48.33 Carrilho and Gilbert, 2000
Porphyridium cruentum 0.08 8.88 Sandau et al., 1996
Spirulina platensis 0.33 37.09 Rangsayatorn et al., 2004
S. platensis 1.07 120.04 Zhou, Huang and Lin, 1998
Spirulina sp. 0.004 0.45 Chojnacka, Chojnacki, and Górecka, 2004
Spirulina sp. 0.10 11.24 Chojnacka, Chojnacki, and Górecka, 2004
S. vulgaris 1.00 112.40 Ofer, Yerachmiel, and Shmuel, 2003
Sargassum baccularia 0.74 83.18 Hashim and Chu, 2004
Sargassum sp. 0.63 70.81 Chaisuksant, 2003
Sargassum sp. 1.07 120.27 Cruz et al., 2004
Sargassum sp. 0.76 85.42 Sheng et al., 2004b
Synechococcus sp. PCC 7942 0.06 7.19 Gardea-Torresdey et al., 1998
Ulva rigida 0.05 5.84 Sandau et al., 1996
Cr (III) Ascophyllum sp. 2.50 129.99 Kratochvil and Volesky, 1998
Ecklonia radiata 2.4 124.79 Kratochvil and Volesky, 1998
Ecklonia sp. 0.74 38.48 Park et al., 2004
Sargassum sp. 2.30 119.59 Kratochvil and Volesky, 1998
Sargassum sp. 1.33 68.89 Cossich, Tavares, and Ravagnani, 2002
Spirulina sp. 0.19 9.62 Chojnacka, Chojnacki, and Górecka, 2005
Synechococcus sp. PCC 7942 0.10 5.41 Gardea-Torresdey et al., 1998
Cr (II) Laminaria japonica 1.81 94.11 Lee et al., 2004
Cr (VI) Padina sp. 1.05 54.60 Sheng et al., 2004b
Pilayella littoralis 0.09 4.68 Carrilho and Gilbert, 2000
Sargassum sp. 0.61 31.72 Sheng et al., 2004b
(Continued on next page)

115 Use of Algae for Removing Heavy Metal Ions From Wastewater
TABLE 1 Recent reports on metal sorption capacity of some algae. (Continued)
a
Sorption

Metal Alga (mmol g−1 ) (mg g−1 ) Reference

Cu Ascophyllum nodosum 0.46 29.29 De Carvalho, Chong, and Volesky, 1995

Ascophyllum sp. 1.26 80.06 Roux, 1998
Chlorella minimata 0.36 23.19 Lau et al., 1999
C. vulgaris 0.17 10.93 Sandau, Sandau, and Pulz, 1996
C. vulgaris 0.29 18.68 Lau et al., 1999
C. vulgaris 1.40 89.02 Mehta and Gaur, 2001a
C. vulgaris 3.00 190.62 Mehta and Gaur, 2001b
C. vulgaris 1.21 76.76 Mehta and Gaur, 2001c
C. vulgaris ( acid pretreated) 6.62 420.63 Mehta, Tripathi, and Gaur, 2002
Durvillaea potatorum 1.30 82.60 Yu and Kaewsarn, 1999
Ecklonia radiata 1.11 70.53 Matheickal and Yu, 1997
E. maxima 1.48 94.0 Feng and Aldrich, 2004
Gracilaria fisheri 0.72 45.75 Chaisuksant, 2003
Laminaria japonica 1.59 101.03 Lee et al., 2004
Microcystis aeruginosa 3.93 249.71 Pradhan et al., 1998
Microcystis sp. 1.57 99.95 Singh, Pradhan, and Rai, 1998
Oscillatoria angustissima 0.12 7.63 Mohapatra and Gupta, 2005
Palmaria palmata 1.05 66.72 Prasher et al., 2004
Pilayella littoralis 0.85 54.01 Carrilho and Gilbert, 2000
Sargassum sp. 0.72 45.75 Chaisuksant, 2003
Spirulina platensis 0.41 25.98 Zhou, Huang, and Lin, 1998
S. platensis 0.16 10.38 Sandau, Sandau, and Pulz, 1996
Spirulina sp. 0.003 0.19 Chojnacka, Chojnacki, and Górecka, 2004
Spirulina sp. 0.19 12.45 Chojnacka et al., 2005
Synechococcus sp. PCC 7942 0.18 11.31 Gardea-Torresdey et al., 1998
Ulva reticulata 0.88 56.3 Vijayaraghavan et al., 2005
Fe Pilayella littoralis 0.70 39.09 Carrilho and Gilbert, 2000
Hg Spirulina sp. 0.007 1.40 Chojnacka, Chojnacki, and Górecka, 2004
Ni Chlorella miniata 0.0007 0.04 Chong, Wong, and Tam, 2000
C. minimata 0.35 20.43 Lau et al., 1999
C. sorokiniana 0.002 0.12 Chong, Wong, and Tam, 2000
C. vulgaris 0.17 10.09 Sandau, Sandau, and Pulz, 1996
C. vulgaris 0.002 0.12 Chong, Wong, and Tam, 2000
C. vulgaris 0.21 12.04 Lau et al., 1999
C. vulgaris 0.02 1.29 Wong, Wong and Tam, 2000
C. vulgaris 1.02 59.70 Dönmenz et al., 1999
C. vulgaris 0.40 23.48 Mehta and Gaur, 2001a
C. vulgaris 3.50 205.48 Mehta and Gaur, 2001b
C. vulgaris (Acid pretreated) 7.46 437.98 Mehta, Singh, and Gaur, 2002
C. vulgaris (Immobilized) 1.89 111.31 Mehta and Gaur, 2001c
Chlorella miniata 0.05 2.99 Wong, Wong, and Tam, 2000
Lyngbya taylorii 0.65 38.16 Klimmek et al., 2001
Microcystis aeruginosa 4.26 249.98 Pradhan et al., 1998
Padina pavonia 1.00 58.71 Ofer, Yerachmiel, and Shmuel, 2003
Pilayella littoralis 0.39 22.89 Carrilho and Gilbert, 2000
S. quadricauda 0.0006 0.04 Chong, Wong, and Tam, 2000
S. vulgaris 1.00 58.71 Ofer, Yerachmiel, and Shmuel, 2003
Sargassum sp. 3.08 180.83 Kalyani, Rao, and Krishnaiah, 2004
Scenedesmus obliquus 0.51 30.18 Dönmenz et al., 1999
Spirulina sp. 0.003 0.18 Chojnacka, Chojnacki, and Górecka, 2004
Synechococcus sp. PCC 7942 0.05 3.17 Gardea-Torresdey et al., 1998
(Continued on next page)

S. K. Mehta and J. P. Gaur 116

TABLE 1 Recent reports on metal sorption capacity of some algae. (Continued)
a
Sorption

Metal Alga (mmol g−1 ) (mg g−1 ) Reference

Synechocystis sp. 3.23 189.81 Dönmenz et al., 1999

Ulva reticulata 0.79 46.51 Vijayaraghavan et al., 2005
Pb C. vulgaris 0.08 17.2 Sandau, Sandau, and Pulz, 1996
Ecklonia radiata 1.36 281.78 Matheickal and Yu, 1996
E. maxima 1.17 243.0 Feng and Aldrich, 2004
Laminaria japonica 1.68 349.09 Lee et al., 2004
Lyngbya taylorii 1.47 304.56 Klimmek et al., 2001
Palmaria palmata 0.07 15.12 Prasher et al., 2004
Schizomeris leibleinni 0.32 65.47 Özer, Özer, and Ekiz, 1999
Spirulina platensis 0.08 16.98 Sandau, Sandau, and Pulz, 1996
Spirulina sp. 0.00005 0.01 Chojnacka, Chojnacki, and Górecka, 2004
Synechococcus sp. PCC 7942 0.15 30.45 Gardea-Torresdey et al., 1998
Zn Ascophyllum nodosum 0.32 20.92 De Carvalho, Chong, and Volesky, 1995
Ascophyllum sp. 1.07 70.01 Roux, 1998
Chlorella miniata 0.009 0.59 Chong, Wong, and Tam, 2000
C. sorokiniana 0.005 0.33 Chong, Wong, and Tam., 2000
C. vulgaris 0.10 6.60 Sandau, Sandau, and Pulz, 1996
C. vulgaris 0.004 0.26 Chong, Wong, and Tam, 2000
Cladophora crispata 0.45 29.42 Özer, Özer, and Dursun, 2000
Laminaria japonica 0.87 56.87 Lee et al., 2004
Lyngbya taylorii 0.49 32.03 Klimmek et al., 2001
Microcystis sp. 9.67 632.12 Singh, Pradhan, and Rai, 1998
Microcystis sp. 15.29 999.50 Pradhan et al., 1998
Oscillatoria anguistissima 0.33 21.57 Mohapatra and Gupta, 2005
O. anguistissima 9.81 641.28 Ahuja, Gupta, and Saxena, 1999
Pilayella littoralis 0.45 29.42 Carrilho and Gilbert, 2000
Sargassum fluitans 1.36 88.96 Leusch, Holan, and Volesky, 1995
S. fluitans 0.85 55.56 Figueira, Volesky, and Ciminelli, 1997
Scenedesmus quadricauda 0.0006 0.04 Chong, Wong, and Tam., 2000
Spirulina platensis 0.11 9.58 Sandau, Sandau, and Pulz, 1996
Spirulina sp. 0.003 0.20 Chojnacka, Chojnacki, and Górecka, 2004
a Not necessarily maximum value.

contribution of the surface-bound Cu declined with a
concomitant increase in intracellular Cu, thereby sug-
gesting the transport of adsorbed Cu into the cells. In-
terestingly, surface adsorption has been invoked as an
important mechanism in algae for tolerating elevated
levels of heavy metals (Lombardi, Vieira, and Sartori,
2002).
Figure 1 shows the probable sites of an algal cell for
the binding of metal ions. Accumulation of metals is
due to adsorption onto the cell surface (wall, mem-
brane or external polysaccharides) and binding to cyto-
plasmic ligands, phytochelatins and metallothioneins,
and other intracellular molecules. Localization of metal
ions on algal cell has been carried out by electron mi-
croscopy and X-ray energy dispersive analysis studies.
Spectroscopy has been used for determining the oxida-
tion state of bound metal on algal cell. Transmission
electron microscopy has shown cell wall as the most
likely location of Cd adsorption by Ectocarpus siliculosus
(Winter, Winter, and Pohl, 1994). Scanning electron mi-
croscopy, in combination with X-ray microanalysis, has
clearly revealed that most of the sites for metal sorption
are present on the surface of algal cells (Klimmek et al.,
2001).
The algal cell wall has many functional groups, such
as, hydroxyl ( OH), phosphoryl ( PO3 O2 ), amino
( NH2 ), carboxyl ( COOH), sulphydryl ( SH), etc.,
which confer negative charge to the cell surface. Since
metal ions in water are generally in the cationic form,
they are adsorbed onto the cell surface (Crist et al.,
1981; Xue, Stumm, and Sigg, 1988; Crist, Martin,
and Crist, 1991; Romero-Gonzalez, Williams, and

117 Use of Algae for Removing Heavy Metal Ions From Wastewater
TABLE 2 Metal sorption capacity of some chemical sorbents.
a
Sorption

Metal Chemical sorbent (mmol g−1 ) (mg g−1 ) Reference

Cd GAC (Hydrodarco 4000) 0.02 2.25 Reed, Arunachalam, and Thomas, 1994
Kaolinite 0.16 17.98 Srivastava et al., 1989
Lignite 0.36 40.46 Allen and Brown, 1995
Montmorillonite 0.36 40.46 Srivastava et al., 1989
Co Duolite C20 1.27 74.84 Kuyucak and Volesky, 1989c
IRA 400 0.37 21.81 Kuyucak and Volesky, 1989c
Cr (III) GAC 0.44 22.88 Leyva-Ramos et al., 1995
Cu Amberlite IRC-718 6.39 406.02 Goto and Goto, 1987
Cation-exchange fibre (IONEX, Toray 2.96 188.08 Goto and Goto, 1987
Industries, Inc.)
Chelite 2.10 133.43 Röhricht, Weppen, and Deckwer, 1990
Duolite 1.80 114.37 Röhricht, Weppen, and Deckwer, 1990
Dowex 50W-X8 5.0 317.7 Goto and Goto, 1987
GAC (BDH-Carbon 330 34, BDH Ltd., Poole, UK.) 0.003 0.191 Scott, Karanjkar, and Rowe et al., 1995
Gellan gum gel 0.75 47.66 Lazaro et al., 2003
Iminodiacetic acid-type chelating fibre 2.84 180.45 Lazaro et al., 2003
IR 120 1.58 101.0 Feng and Aldrich, 2004
IR C467 1.26 80 Feng and Aldrich, 2004
Lewatite 2.80 177.91 Röhricht, Weppen, and Deckwer, 1990
Hg Lewatite MP 64 1.99 399.17 Ferreira and Carvalho, 1997
Lewatite TP 214 1.99 399.17 Ferreira and Carvalho, 1997
Lignite 0.38 76.22 Allen, 1995
Pb Cation exchange resin with sulfonyl group 0.80 165.75 Suh and Kim, 2000
(SK 1B, Sam Yang Co., Korea)
GAC (Sigma, USA) 0.018 3.73 Suh and Kim, 2000
Gellan gum gel 0.85 176.11 Lazaro et al., 2003
Hydrodarco 4000 (American Norit Company, 0.027 5.594 Reed, Arunachalam, and Thomas, 1994
Inc.
Zn Lignite 0.38 24.84 Allen, 1995
a Not necessarily maximum value.

Gardiner, 2001; Skowroñski and Ska, 2000). Each func-
tional group has a specific pKa (dissociation constant)
(Eccles, 1999; Niu and Volesky, 2000), and it disso-
ciates into corresponding anion and proton at a spe-
cific pH. These functional groups are found associated
with various cell wall components, e.g., peptidoglycan,
teichouronic acid, teichoic acids, polysaccharides and
proteins. Because distribution and abundance of cell
wall components vary among different algal groups,
the number and kinds of functional group also vary
in different algal groups. Among different cell wall con-
stituents, polysaccharides and proteins have most of the
metal binding sites (Kuyucak and Volesky, 1989a).
Peptidoglycan, consisting of linear chains of
N-acetylglucosamine and β 1, 4-N-acetylmuramic acid
with peptide chain, is the major component of cell
wall of cyanobacteria (blue-green algae), providing
mostly carboxylic groups for metal binding (Chojnacka,
Chojnacki, and Górecka, 2005). Lipopolysaccharides,
lipids and membrane proteins are also important in
metal binding by cyanobacteria. Phosphoryl groups
are mainly associated with lipopolysaccharides, lipids
and peptidoglycans of cell wall. Amino groups are as-
sociated with membrane proteins and peptide com-
ponent of peptidoglycan. It has been suggested that
carboxyl groups on the cyanobacterial cell wall are
the dominant active sites for the binding of metal
ions (Chojnacka, Chojnacki, and Górecka, 2005). Many
strains of cyanobacteria have an outer sheath or capsule
made up of polysaccharides. Capsule of cyanobacteria
is anionic in nature, owing to presence of uronic acids
and/or of other charged groups (De Philippis and Vin-
cenzini, 1998). Due to its anionic nature cyanobacterial
capsule usually shows very high affinity towards metal

S. K. Mehta and J. P. Gaur 118

FIGURE 1 Metal binding sites of a typical algal cell. The alphabet M represents the metal species (independent of its oxidation state).
ions and may offer a promising chelating agent for re-
moval of heavy metals from wastewaters (De Philippis
et al., 2001). A wide difference in metal binding capacity
of capsulated and uncapsulated cyanobacterial strains
has been demonstrated (Pradhan and Rai, 2000). A
cyanobacterial strain with thick capsule showed higher
metal binding capacity than strains devoid of or with a
thin layer of capsule (Singh, Pradhan, and Rai, 1998; De
Philippis et al., 2003). However, presence of gelatinous
capsule around cyanobacterial cell may slow down the
diffusion rate of metal ions into the chelating matrix of
cell wall (De Philippis et al., 2003).
Cell wall of green algae contains heteropolysaccha-
rides, which offer carboxyl and sulfate groups (Lee,
1980) for sequestration of heavy metal ions. The ex-
tracted polysaccharides (12% of dry weight) from Ulva
sp. contained 16% sulfate and 15–19% uronic acid
(McKinnel and Percival, 1962). Protein content of cell
wall of green algae ranges from 10 to 70% (Siegel and
Siegel, 1973). Aspartic and glutamic acid account for
12% of protein content of cell wall, which correspond to
0.15 meq g−1 of carboxylic groups per dry weight. In the
cell wall of green algae, lysine and arginine make up 13%
of protein (∼0.08 meq g−1 of amino groups) (Chapman,
1980; Lee, 1980; Guing and Blunden, 1991).
119
Marine algae have been the focus of numerous
biosorption studies and their excellent metal binding
capacity has been widely acknowledged. The main con-
stituents of the cell wall of brown algae are cellulose,
as the fibrous skeleton, and alginate and fucoidan that
constitute the amorphous matrix, and extracellular mu-
cilage (Lee, 1980). It is fairly well recognized that al-
ginate is mainly involved in metal accumulation by
brown algae (Kuyucak and Volesky, 1989a). Alginate is
defined as the ammonium or alkali salt of alginic acid.
Following a mild acid hydrolysis of alginic acid, three
kinds of segments are demonstrated as building blocks
of the polymer. These are D-mannuronic acid and
L-guluronic acid units, and alternating D-mannuronic
acid and L-guluronic acid residues (Davis, Volesky, and
Mucci, 2003). The carboxyl groups of each polymer
segment may play an important role as the site for
cation binding. Polyguluronic acid shows a high speci-
ficity for divalent metal ions (Puranik, Modak, and
Paknikar, 1999). Haug (1967) showed that the affinity
of alginates for divalent cations such as Pb2+ , Cu2+ ,
Cd2+ , Zn2+ , Ca2+ , etc. increased with guluronic acid
content. Alginate content of brown algae is 10 to
40% of dry weight (Percival and McDowell, 1967). Al-
ginate concentration in Sargassum fluitans is 45% of
Use of Algae for Removing Heavy Metal Ions From Wastewater
its dry weight, corresponding to 2.25 mmol of car-
boxyl groups g−1 of biomass (Fourest and Volesky,
1996). Calpomenia has 5–14% alginate (∼0.25–0.7 meq
g−1 carboxyl groups) (Kalimuthu, Kaliaperumal, and
Ramalingam, 1991). Brown algae also contain about
5 to 20% sulfated polysaccharide fucoidan (Chapman,
1980), about 40% of which is sulfate esters. Fucoidan is
a branched polysaccharide sulfate ester with L-fucose
builing blocks, which are predominantly α(1 → 2)
linked. Uptake of trivalent cations has been attributed
mainly to the presence of this sulfated polysaccha-
ride (Figueira, Volesky, and Mathieu, 1999). About
0.27 ± 0.03 meq g−1 sulfonate groups were reported in
Sargassum fluitans (Fourest and Volesky, 1996). These lat-
ter authors showed that the contribution of sulfonate
to heavy metal binding is generally small, but may be
significant at low pH. Protein (10% of biomass) con-
tributes ∼0.17 meq/g carboxyl groups to Sargassum
biomass. Lysine and arginine (10% of protein) offer
∼0.07 meq/g amine group. In brown algae, the car-
boxyl group of alginate is more abundant than either
carboxyl or amine groups of protein.
Functional groups involved in metal sorption by al-
gae have been identified, but not fully, by FTIR spec-
troscopy, pH-titration, potentiometric and conducti-
metric titration techniques, and also after blocking of
functional groups with certain chemical agents. FTIR
analysis led Ting, Teo, and Soh (1995) to suggest the
involvement of carboxyl groups in binding of Au (III)
on Chlorella vulgaris. Participation of carboxyl groups
in adsorption of Cu, Cd and Pb on cyanobacterial
cell wall has also been demonstrated (Yee et al., 2004).
Seki and Suzuki (1998) showed that biosorption of bi-
valent metal ions by brown algae, namely, Macrocys-
tis pyrifera, Kjellmaniella crassiforia and Undaria pinnati-
fida, was due to binding to carboxyl groups of alginic
acid. On the basis of potentiometric titration, 13 C-
nuclear magnetic resonance (NMR), and chemical anal-
ysis, Fourest and Volesky (1997) have demonstrated that
heavy metal binding capacities of four different brown
seaweeds (S. fluitans, Ascophyllum nodosum, Fucus vesicu-
losus, and Laminaria japonica) were directly proportional
to the number of carboxyl groups. Romero-Gonzalez,
Williams, and Gardiner (2001) found up to 95% de-
crease in Cd biosorption by dealginated seaweed waste
after esterification of biomass, and thus concluded that
carboxyl groups are largely responsible for Cd biosorp-
tion. Furthermore, using FTIR analysis of non-living S.
fluitans, Fourest and Volesky (1996) showed binding of
S. K. Mehta and J. P. Gaur
Cd and Pb largely on the carboxyl and to a small extent
on sulfonate groups. Mehta, Singh, and Gaur (2002)
demonstrated the involvement of carboxyl groups in
adsorption of Cu onto C. vulgaris. Seki, Suzuki and Iburi
(2000) suggested that sorption of bivalent metal ions
by a marine microalga Heterosigma akashiwo involves
monodentate binding on carboxylic- and phosphatic-
type sites. A significant role of carboxyl groups in sorp-
tion of heavy metals has also been very well demon-
strated in fungi as well as in higher plants (Kapoor and
Viraraghavan, 1997; Aksu and Dönmez, 2001).
Participation of sulfonate, amino and hydroxyl
groups in adsorption of various metal ions has been
demonstrated, but to a lesser extent than that of car-
boxyl groups. On the basis of IR spectroscopy of non-
living Sargassum natans, Kuyucak and Volesky (1989a)
demonstrated the binding of Co on carbonyl groups.
Potentiometric titration study on Chlamydomonas rein-
hardtti revealed the presence of amino and thiol groups
along with carboxyl groups. According to Greene et al.
(1986), amino group plays an important role in bind-
ing of Au to C. vulgaris as pretreatment of the alga with
succinic anhydride, that is known to react with amino
groups making them unavailable for metal binding, re-
sulted in a significant (50%) decrease in Au binding. On
the basis of an IR data, Gong et al. (2005) concluded
that amino and hydroxyl groups play a predominant
role (at high pH) in binding of Pb on Spirulina maxima.
Ting, Teo and Soh (1995) studied the mechanism of
Au (III) sorption by C. vulgaris using X-ray photoelec-
tron spectroscopy (XPS) analysis. They found elemental
gold on the cell surface, and this led them to suggest
the involvement of a reduction process in Au sorption.
Greene et al. (1986) suggested that sulphydryl groups are
not involved in Au binding by Chlorella vulgaris.
The above discussion leads to the generalization that
carboxyl groups of cell wall polysaccharides play a pre-
dominant role in heavy metal sorption by algae and
cyanobacteria. The other functional groups, like sul-
fonate and amino, play a relatively minor role in metal
sorption. Thiol group plays an important role in sorp-
tion of metals like Cd at lower pH (>2) (Sheng et al.,
2004a). Although various potential fuctional groups
have been demonstrated in algae, their mere presence
does not guarantee participation in biosorption of met-
als. Steric hinderance, conformational changes or cross-
linking, all of which change with environmental condi-
tions (pH, ionic strength, competing cations or ligands),
may prevent some surface functional groups from
120
binding to metal (Adhiya et al., 2002). For example, sur-
face charge may be neutralized either by binding with
cations or by cross-linking between oppositely charged
surface groups.
Recent studies reveal that the process of heavy
metal biosorption involves mechanisms, such as ion
exchange, complexation, electrostatic attraction and
microprecipitation (Volesky and Holan, 1995). Ion
exchange has been shown to be the most important
mechanism for the biosorption of metal ions by algal
biomass (Crist et al., 1990). Greene, McPherson, and
Darnall (1987) observed that the binding of Cu2+ , Pb2+ ,
Zn2+ , Ni2+ and Cr3+ to Spirulina platensis was accom-
panied by the liberation of protons from the biomass.
Release of Mg2+ after binding of Cu2+ and Zn2+ on
Oscillatoria anguistissima has been reported by Ahuja,
Gupta and Saxena (1999). Williams and Edyvean (1997)
observed a concomitant release of Ca2+ during biosorp-
tion of Ni2+ by the brown alga Ecklonia maxima. All
these studies suggest that heavy metal sorption involves
exchange of metal ions with surface bound protons or
cations. Ion exchange capacity is highly variable among
different algal species. Pirszel, Pawlik, and Skowroñski
(1995) reported the maximum ion exchange capacity (in
term of Na+ /H+ exchange) of five algal species ranging
from 41–825 µeq g−1 dry weight. Large variability in
ion exchange capacity of different algal forms may be
due to variation in cell wall composition. Unicellular al-
gae generally show a higher ion exchange capacity than
filamentous forms due to higher surface/volume ratio
(Pirszel, Pawlik, and Skowroñski, 1995). Ion exchange
capacity and consequently metal sorption capacity, of
algal biomass increases with an increase in pH (Pirszel,
Pawlik, and Skowroñski, 1995; Mehta, Singh and Gaur,
2002). This has been explained as an effect of decreas-
ing competition with protons for the same binding sites
(Crist et al., 1994). Some researchers believe that other
mechanisms, like complexation (or coordination), are
important in metal sorption by algae (Davis, Volesky,
and Mucci, 2003). Raiz, Argaman, and Yannai (2004)
studied the mechanism of metal binding by Sargassum
vulgaris and reported that Cd binding involves chela-
tion, whereas binding of Pb is mediated by a combina-
tion of ion exchange and chelation. Adhiya et al. (2002)
reported that Cd biosorption to Chlamydomonas rein-
hardtti involves complexation with carboxylic groups.
Electrostatic attraction and covalent binding, respec-
tively, mediate Ni and Zn adsorption on Chaetophora
elegans (Andrade, Rollemberg, and Nóbrega, 2005). In
121
view of complexicity of the composition of the algal
surfaces, it is possible that various mechanisms oper-
ate simultaneously to varying degrees depending on al-
gal species and the environmental conditions (Sheng
et al., 2004a). Aluminum sorption onto algal cells in-
volves a different kind of mechanism. Al ions bind to
biomass in the form of polynuclear Al species such
as [Al6 (OH)12 (H2 O)12 ]6+ (Hsu and Bates (1964) and
Al13 (OH)7+32 (Bottero et al., 1980). These polymerized
Al ions prevented other heavy metal ions from access-
ing to binding site.
Once metals are inside the cell, they may bind to
intracellular components or precipitate (Gadd, 1988).
Biological macromolecules and enzymes with appropri-
ate functional groups or metal co-factors are impacted
by metal activity. In Ankistrodesmus falcatus, Wong et al.
(1984) have demonstrated that Sn was about 85% in
the cellular polysaccharide fraction, 15% in the protein
fraction, and 0.2% in lipid and low molecular weight
fractions. Metals may be detoxified by accumulation
in polyphosphate bodies and in intracellular metal-
binding proteins of cyanobacteria and eukaryotic algae
(Zhang and Majidi, 1994), and within the vacuoles of
some eukaryotic algae (Gadd, 1988; Garnham, Codd,
and Gadd, 1992). The presence of one metal may change
the distribution of others among cellular components.
Okamura and Aoyama (1994) have shown that when
present in a mixture, Cd and Cr (VI) influence each
other’s concentration and distribution among mem-
brane, cell wall, and soluble and miscellaneous fractions
of Chlorella ellipsoidea.
Algal cells have considerable potential in adsorbing
anionic species of certain metals. For instance, algal
cells can also adsorb Cr (VI) with considerable ease at
low pH values (<2) (Kratchovil, Pimentel, and Volesky,
1998; Dönmez and Aksu, 2002). Removal of Cr (VI)
by algae is anionic as well as through its reduction to
the cationic Cr (III) under strongly acidic conditions.
In nature Cr occurs in these two forms only, with Cr
(VI) being much more toxic to biota than Cr (III). At
low pH, the algal biomass provides protons for the re-
duction of Cr (VI) to Cr (III). Protonated biomass of
Ecklonia, a brown seaweed, could completely reduce Cr
(VI) to Cr (III) (Park et al., 2004). Cr (III) thus formed
was present in the solution or as partly bound to the
biomass. Cr (VI) reduction was independent of biomass
concentration but it increased with decreasing pH. In
another paper, Park et al. (2004) used prolonged (1 or
10 days) thermal treatment (60–100◦ C) for enhancing
Use of Algae for Removing Heavy Metal Ions From Wastewater
the capability of Ecklonia biomass in reducing Cr (VI)
to Cr (III). However, thermal treatment decreased the
number of carboxyl groups by 76% and this inter alia de-
creased the efficiency of Cr (III) adsorption. The latter
authors suggested a two-stage process for the complete
removal of Cr (VI) and Cr (III), using thermally treated
biomass for reducing Cr (VI) at lower pH and subse-
quently using native biomass for removing Cr (III) at
higher pH.
III. METAL REMOVAL BY ALGAL
BIOSORPTION
Limited efforts have been made to use algal biomass
for removing metal ions from aqueous solutions. Al-
though thousands of algal species are known, only a
few of them have been investigated for their metal
sorption ability and subsequent use in wastewater treat-
ment. Metal biosorption experiments have been con-
ducted with freshwater green algae (e.g., Chlorella spp.,
Cladophora spp., Scenedesmus spp., Chlamydomonas rein-
hardtii), brown algae (e.g., Sargassum natans, Fucus vesicu-
losus, Ascophyllum nodosum, Laminaria japonica), and
blue-green algae (like Microcystis aeruginosa and Oscillato-
ria). Microalgae are easy to grow in culture and some al-
gal species are being grown commercially in large quan-
tity. Metal sorption ability of algae varies greatly from
species to species and even among strains of a single
species for any metal (Table 1), although this variation
may also be due to variable experimental conditions
in different studies. A suggestion has also been made
that cells grown under different conditions vary with
regard to composition of their cell wall, and hence in
biosorption characteristics (Chojnacka, Chojnacki, and
Górecka, 2005). Some algae show a high affinity for
sorbing a particular metal ion, whereas others do not
show such specificity and may sorb several metal ions.
The affinity of various algal species for binding of
metal ions shows different hierarchies. In general, metal
ions with greater electronegativity and smaller ionic
radii are preferentially sorbed by algal biomass. The
available literature suggests that Pb is sorbed maximally
compared to other metals in a majority of algal species
(Tiem, 2002; Davis, Volesky, and Mucci, 2003). Brown
algae like Ascophyllum, Sargassum, etc., sorb more metal
than other algae due to their high alginate content. In
general, metal sorption capacity of algae is the least for
Ni, although M. aeruginosa possesses a very high Ni
sorption capability (Pradhan et al., 1998). Lee et al. (2000)
S. K. Mehta and J. P. Gaur
screened 48 species of red, brown and green marine al-
gae for their Cr (VI) adsorbing potential, and found
extraordinarily high selectivity of Cr (VI) sorption in
Pachymeniopsis sp. (Rhodophyta). This alga was found
to be poor in sorbing other heavy metals from water.
Metal sorption potential of brown algae has been more
thoroughly investigated compared to that of other ma-
rine algae and freshwater filamentous algae. Freshwater
filamentous diatoms and sheath-forming filamentous
blue-green algae, that might bind heavy metals with
great efficiency, have somehow failed to receive ade-
quate attention of the researchers.
The metal removal ability of algae is comparable
and even at times higher than that of other sorbents
(Table 3). Metal removal efficiency of the commonly
used ion exchangers and resins is very low at or below
10 mg l−1 metal concentration in the solution. Nev-
ertheless, algae may almost completely remove metal
ions from solutions having low metal concentrations
(Mehta and Gaur 2001a; Mehta, Tripathi, and Gaur,
2002). Axtell, Sternberg and Claussen (2003) found
97% removal of Pb by the macroalga Macrospora from
a solution having 39.4 mg l−1 initial concentration
of Pb. Many times algae have been found to outper-
form other biosorbents in removing metals from so-
lutions. Algal biomass could be used in the form of
biotraps for the removal of heavy metals from indus-
trial effluents. AlgaSORB
R , a commercial product, con-
sisting of gel encapsulated algal cell wall, has a re-
markable affinity for Hg, Pb, Cd, Cr, Cu, Zn, Ni,
Ag, Au, etc. (Darnall, 1989). An important charac-
teristic of AlgaSORB
R is that high concentrations of
common ions do not interfere with sorption of metal
ions. Where synthetic resins are inefficient in removing
heavy metals from wastewaters, having high concen-
trations of total dissolved solids (TDS), AlgaSORB
R
could be used successfully to remove toxic metals
(Darnall, 1991). Chitoplex, an insoluble cross-linked
chitosan, is another commercial biotrap for detoxifica-
tion of heavy metal-containing industrial wastewaters
(Crusberg, Weathers, and Baker, 1991).
A. Factors Affecting Metal
Sorption by Algae
Biosorption of heavy metals by algae may be affected
by several factors, including concentration of metal and
biomass, pH, temperature, presence of competing ions
and the metabolic stage of the organism.
122
TABLE 3 Recent reports on metal removal efficiency of algae and some other materials.
Initial Metal
concentration removal
of metal efficiency
Metal sorbent Metal (mg l−1 ) (%) Reference
c
Anabaena subcylindrica Cu IE 82 El-Sheekh et al., 2005
A. subcylindrica Co IE 34 El-Sheekh et al., 2005
A. subcylindrica Pb IE 100 El-Sheekh et al., 2005
A. subcylindrica Mn IE 100 El-Sheekh et al., 2005
Ascophyllum nodosum Cd 500 98 Sandau et al., 1996
Chlorella vulgaris Cu 10 72 Mehta and Gaur, 2001a
C. vulgaris Cu 2.5 80 Mehta and Gaur, 2001a
C. vulgaris Ni 2.5 69 Mehta and Gaur, 2001a
C. vulgaris Cu 10 72 Mehta and Gaur, 2001
C. vulgaris — 48 Travieso et al., 1999
C. vulgaris Cd 500 84 Sandau et al., 1996
C. vulgaris (acid-pretreated) Cu 2.5 96 Mehta and Gaur, 2001a
C. vulgaris (HCl-pretreated) Ni 2.5 93 Mehta and Gaur, 2001a
C. vulgaris (Ca-alginate immobilized) Cu 5.0 >90 Mehta and Gaur, 2001c
C. vulgaris (Ca-alginate immobilized) Ni 5.0 70 Mehta and Gaur, 2001c
De-alginated seaweed Cd 10 91 Romero-Gonzalez, Williams, and Gardiner, 2001
Fucus vesiculosus Cd 500 98 Sandau et al., 1996
Laminaria digitata Cd 500 98 Sandau et al., 1996
Nostoc muscorum Cu IE 13 El-Sheekh et al., 2005
N. muscorum Co IE 12 El-Sheekh et al., 2005
N. muscorum Pb IE 26 El-Sheekh et al., 2005
N. muscorum Mn IE 33 El-Sheekh et al., 2005
Pachymeniopsis sp. Cr 200 57 Lee et al., 2000
Phormidium valderianum Cd 40 mM 90 Karna et al., 1999
Sargassum sp. Zn 98 99 Esteves, Valdman, and Leite, 2000
Sargassum sp. 162 52 Cossich, Tavares, and Ravagnani, 2002
Sargassum sp. Cd 3 100 Esteves, Valdman, and Leite, 2000
Scenedesmus abundans Cd 10 97 Terry and Stone, 2002
S. abundans Cu 10 99 Terry and Stone, 2002
S. quadricauda Ni 30 97 Chong, Wong, and Tam, 2000
Spirulina platensis Cd 500 81 Sandau et al., 1996
Spirulina sp. Al — 49 Chojnacka, Chojnacki, and Górecka, 2004
Spirulina sp. Cd — 56 Chojnacka, Chojnacki, and Górecka, 2004
Spirulina sp. Co — 21 Chojnacka, Chojnacki, and Górecka, 2004
Spirulina sp. Cu — 48 Chojnacka, Chojnacki, and Górecka, 2004
Spirulina sp. Hg — 60 Chojnacka, Chojnacki, and Górecka, 2004
Spirulina sp. Ni — 22 Chojnacka, Chojnacki, and Górecka, 2004
Spirulina sp. Pb — 95 Chojnacka, Chojnacki, and Górecka, 2004
Spirulina sp. Zn — 67 Chojnacka, Chojnacki, and Górecka, 2004
Spirulina maxima Pb — 92 Gong et al., 2005
S. maxima (CaCl2 pretreated) Pb — 84 Gong et al., 2005
Tetraselmis suecica Cd 45 60 Perez-Rama et al., 2001
Alginate Ni 10 20 Mehta and Gaur, 2001c
Alginate Cu 10 40 Mehta and Gaur, 2001c
Alginate Cd 500 72 Sandau et al., 1996
Agar-Agar Cd 500 47 Sandau et al., 1996
Chitin Cd 500 31 Sandau et al., 1996
Chitosan Cd 500 76 Sandau et al., 1996
Sphagnum∗ Cr 100 100 Sharma and Forster, 1995
(Continued on next page)

123 Use of Algae for Removing Heavy Metal Ions From Wastewater
TABLE 3 Recent reports on metal removal efficiency of algae and some other materials. (Continued)
Initial Metal
concent-ration removal
of metal efficiency
Metal sorbent Metal (mg l−1 ) (%) Reference

Aspergillus terreus Cd 10 70 Massaccesi et al., 2002

Cladosporium cladosporiodes Cd 10 63 Massaccesi et al., 2002
Talaromyces helicus Cd 10 70 Massaccesi et al., 2002
Trichoderma koningii Cd 10 64 Massaccesi et al., 2002
a
Yeast, granulated Zn EF <99 Brady, Stoll and Duncan, 1994
a
Yeast, granulated Cr EF <99 Brady, Stoll and Duncan, 1994
a
Yeast, granulated Cu EF <99 Brady, Stoll and Duncan, 1994
b
Yeast, granulated Cr TE 60 Brady, Stoll and Duncan, 1994
Dry Yeast Cd 500 73 Sandau et al., 1996
Saccharomyces cerevisiae Cd 500 36 Sandau et al., 1996
S. cerevisiae Cu 50 µM 49 Huang, Huang and Morehart, 1990
Apple pomace, dry Cd 500 62 Sandau et al., 1996
Wood dust, dry Cd 500 37 Sandau et al., 1996
Escherichia coli (engineered) Zn 500 µM 99 Bang, Clark and Keasling, 2000
E. coli (engineered) Pb 200 µM 99 Bang, Clark and Keasling, 2000
E. coli (engineered) Cd 200 µM 91 Bang, Clark and Keasling, 2000
Fe(III)/Cr(III) hydroxide Pb 200 95 Namasivayam and Ranganathan, 1995
Fe(III)/Cr(III) hydroxide Cd 80 86 Namasivayam and Ranganathan, 1995
Fe(III)/Cr(III) hydroxide Ni 50 82 Namasivayam and Ranganathan, 1995
a EE = Effluent from electroplating industry; b TE = Effluent from tannery; c IE = Industrial effluent.

1. Initial Metal Ion Concentration
Sorption and removal of heavy metals by algal
biosorbents largely depend on the initial concentration
of metals in the solution. Metal sorption initially in-
creases with increase in metal concentration in the so-
lution, and then becoming saturated after a certain con-
centration of metal (Da Costa and Leite, 1991; Aloysius,
Karim, and Arif, 1999; Mehta and Gaur, 2001a,b,c;
Mehta, Singh, and Gaur, 2002; Mehta, Tripathi and
Gaur, 2002). Algal cell surface has several kinds of func-
tional groups with varying affinity for an ionic species.
Low and high affinity functional groups are involved
in sorption of metal ions at high and low concentra-
tions of metal ions, respectively. In contrast to sorp-
tion, the removal of a metal generally decreases with
its increasing concentration in the solution (Mehta and
Gaur, 2001a,c). The latter authors found that Chlorella
vulgaris could remove 70 and 80% Ni and Cu, respec-
tively, from their 2.5 mg l−1 solutions; however, only 37
and 42% Ni and Cu, respectively, were removed from
their 10 mg l−1 solutions.
2. pH
Most of the studies have shown that sorption of
metal ions in batch as well as in continuous system is a
function of pH of the solution. Therefore, efforts have
been made to find out optimum pH for maximizing
metal removal by algae. The sorption of Cr (VI) and Cd
on Padina sp. and Sargassum sp. (Sheng et al., 2004b),
and Cs sorption on Padina australis (Jalali-Rad et al.,
2004) was optimal at pH 2. Yu and Kaewsarn (1999)
found very little sorption of Cu by Durvillaea potato-
rum at pH below 2, but it increased with a rise in pH.
They found maximum Cu sorption between pH 3 and
4, and the plateau was reached at around pH 5. There
are numerous studies showing increased metal sorption
with increasing pH of the solution. Zhou, Huang, and
Lin (1998) suggested that the optimum pH for Cu and
Cd sorption by Laminaria japonica and Sargassum kjell-
manianum lies between 4 and 5. Cossich, Tavares and
Ravagnani (2002) found maximal Cr (III) sorption ca-
pacity of Sargassum sp. at pH 4. Some studies show selec-
tive sorption of specific metals due to widely distinct pH
optima for their sorption. Özer et al. (1994) showed op-
timum sorption of Pb and Cr (IV) by Cladophora crispata
at pH 5.0 and 1.0, respectively. The different optimum
pH for the above two metals could be due to variable
nature of their chemical interaction with the algal cell.
At pH 5.0, cells have a net negative charge on the surface
that favors the binding of Pb (II) to surface ligands. At

S. K. Mehta and J. P. Gaur 124

pH below iosoelectric point, cells have a positive charge
that inhibits the approach of positively charged Pb (II)
ions. Because Cr (IV) is anionic in nature, it does not
bind at high pH when the overall algal surface charge
is negative. Cr (IV) and other anions bind to algal sur-
face at low pH when algal surface is positively charged.
In general, acidic pH (3–5) is most favorable for the
sorption of metal ions (Table 4).
Since a majority of the metal binding groups of al-
gae are acidic (e.g., carboxyl), their availability is pH-
dependent. These groups generate a negatively charged
surface at acidic pH, and electrostatic interactions be-
tween cationic species and the cell surface is responsible
for metal biosorption. A decreased metal sorption by
algae has been frequently observed at extremely acidic
pH (<2) (Crist et al., 1981; Özer et al., 1994; Mehta and
Gaur, 2001a). High concentration of H+ ions at low
pH decrease metal sorption by competitively excluding
them from binding to ligands on the cell surface (Pe-
terson, Healey, and Wagemann, 1984; Campbell and
Stokes, 1985; Gensemer, 1991; Parent and Campbell,
1994). Table 4 also shows great variability in optimum
pH for sorption of a particular metal ion by different al-
gal species. For example, the optimum pH for the sorp-
tion of Cu by blue-green algae (Microcystis aeruginosa
and Spirulina platensis) is far greater than that for Cu
sorption by green algae (Cladophora prolifera, Chlorella
vulgaris, C. kessleri). Similarly, optimum pH for Cd sorp-
tion is 4.0 (Esteves, Valdman, and Leite, 2000) and
7.0 for Sargassum sp. and Stichococcus bacillaris, respec-
tively. This variability may be related to the differences
in chemical composition of cell surface of various al-
gal species. Chojnacka, Chojnaki, and Gorecka (2005)
[Mn+ ] 1
=

 n

−
+ βnCl [Cl− ]n + βnSO4 SO4
[M]T n 2−
1 + βnOH [OH ] + βnCO3 CO3
noted a distinct relationship between pH of aqueous
metal solution and involvement of fuctional group in
binding of Pb onto Spirulina maxima; for pH ranges
2–5, 5–9 and 9–12, respectively, the functional groups
involved in binding of Pb were carboxyl, carboxyl and
phosphate, and carboxyl, phosphate and hydroxyl.
3. Metal Speciation
A large influence of pH on metal sorption can also be
related to the availability of free metal ions in test solu-
125
tion. Metals in wastewaters occur in a variety of chemi-
cal forms, namely, free aquo ions, complexed with inor-
ganic and organic ligands, and adsorbed on particulate
phases. Indeed, toxic effects of metal ions to organisms
depend on the concentration of free aquo ions (Sunda
and Guillard, 1976; Anderson and Morel, 1982; Sunda
and Ferguson, 1983). Likewise, sorption of metals is also
a function of concentration of free metal ions in the so-
lution (Volesky and Holan, 1995). Availability of metal
ions for binding onto algae depends on chemical specia-
tion, which in turn is determined by pH of the solution.
Wastewaters may have inorganic ligands like HCO− 3,
CO2− , Cl − , SO2− , HS− , and organic compounds, such
3 4
as acetic acid, oxalic acid, amino acids, etc., for metal
binding. Metal ions in a natural water sample may also
be bound on biological materials and different kinds
of solid surfaces. The free metal ion concentration is
regulated by complex interactions between metal ions,
ligands and major ions. It is also regulated by pH of
the solution, and the extent of hydrolysis and stability
constant (β) of metal-ligand complexes (Sigg and Xue,
1994). The concentrations of free ions of various met-
als like Pb (II), Cu (II), Hg (II), Al (III) and Fe (III)
in freshwater remain very low in the pH range 7.5–9.0
due to complexation with inorganic ligands (Sigg and
Xue, 1994). Decrease in pH of the solution generally
increases the concentration of free metal ions. For an
efficient removal of heavy metals by an algal biosor-
bent, the ratio of free metal ion to total metal concen-
tration should remain high. The ratio of free metal ion
[Mn+ ] to total metal concentration [M]T in a solution
can be determined by the free ligand concentration and
stability constant (β) (Sigg and Xue, 1994):

 2− n
(1)
In this equation, only inorganic ligands like OH− ,
CO23 , Cl− and SO2− 4 have been considered. Using
the above model, Sigg and Xue (1994) listed some
major species and [Mn+ ]/[M]T for some elements in
freshwater at pH 8. They have shown that at this pH
[Mn+ ]/[M]T for Fe (III), Co (II), Cu (II), Zn (II), Pb
(II) and Hg (II) was very low, i.e., 2 × 10−11 , 0.5, 0.4,
0.01, 0.4, 5 × 10−3 and 1 × 10−10 , respectively. Most of
the studies have ignored this important aspect during
screening of algal species for sorption of metals from
metal solutions or industrial effluents.
Use of Algae for Removing Heavy Metal Ions From Wastewater
TABLE 4 Optimum pH for sorption of heavy metals by algae.
Metal Alga pH Reference

Ag Cladophora prolifera 5.0 Hao, Zhao, and Ramelow, 1994

Oedogonium sp. 7.0 Crist et al., 1988
Padina pavonica 4.0 Hao et al., 1994
Ulva lactuca 5.0 Hao, Zhao, and Ramelow, 1994
Al Cladophora prolifera 4.0 Hao, Zhao, and Ramelow, 1994
Padina pavonica 5.0 Hao, Zhao, and Ramelow, 1994
Pilayella littoralis 5.5 Carrilho and Gilbert, 2000
Ulva lactuca 5.0 Hao, Zhao, and Ramelow, 1994
Au Cladophora prolifera 2.0 Hao, Zhao, and Ramelow, 1994
Padina pavonica 4.0 Hao, Zhao, and Ramelow, 1994
Sargassum sp. 2.0 Niu and Volesky, 2000
Ulva lactuca 3.0 Hao, Zhao, and Ramelow, 1994
Cd Cladophora prolifera 6.0 Hao, Zhao, and Ramelow, 1994
Fucus vesiculosus 3.5–6.8 Sandau et al., 1996
Laminaria japonica 4.0–5.0 Zhou, Huang, and Lin, 1998
L. japonica 6.0 Yin et al., 2001
Lyngbya taylorii 3–7 Klimmek et al., 2001
Padina pavonica 6.0 Hao, Zhao, and Ramelow, 1994
Pilayella littoralis 5.0 Carrilho and Gilbert, 2000
Sargassum kjellmanium 4.0–5.0 Zhou, Huang, and Lin., 1998
Sargassum sp. 4.5 Esteves, Valdman, and Leite, 2000
Seaweed waste (dealginated) 6–8 Romero-Gonzalez et al., 2001
Stichococcus bacillaris 7.0 Skowroñski, 1986
Ulva lactuca 6.0 Hao, Zhao, and Ramelow, 1994
Co Ascophyllum nodosum 4.0–5.0 Kuyucak and Volesky, 1989a
Cladophora crispata 1.0 Özer et al., 1994
C. prolifera 6.0 Hao, Zhao, and Ramelow, 1994
Padina pavonica 6.0 Hao, Zhao, and Ramelow, 1994
Pilayella littoralis 5.5 Carrilho and Gilbert, 2000
Cr (VI) Chlorella vulgaris 2.0 Aksu and Acikel, 1999
Cladophora crispata 1.0 Özer et al., 1994
Padina sp. 2.0 Sheng et al., 2004b
Sargassum sp. 2.0 Sheng et al., 2004b
Cr (III) Sargassum sp. 4.0 Cossich, Tavares, and Ravagnani, 2002
C. prolifera 3.0 Hao, Zhao, and Ramelow, 1994
Padina pavonica 5.0 Hao, Zhao, and Ramelow, 1994
Ulva lactuca 3.0 Hao, Zhao, and Ramelow, 1994
Cu Chlorella vulgaris 4.0 Aksu and Acikel, 1999
C. vulgaris 3.5 Mehta and Gaur, 2001a
Cladophora prolifera 4.0 Hao, Zhao, and Ramelow, 1994
Laminaria japonica 5.0 Zhou, Huang, and Lin., 1998
Microcystis aeruginosa 7.0 Pradhan, et al., 1998
Padina pavonica 5.0 Hao, Zhao, and Ramelow, 1994
Pilayella littoralis 5.5 Carrilho and Gilbert, 2000
Sargassum kjellmanium 4.0–5.0 Zhou, Huang, and Lin, 1998
Spirulina platensis 6.7 Zhou, Huang, and Lin, 1998
Ulva lactuca 5.0 Hao, Zhao, and Ramelow, 1994
U. lactuca 6.0 Hao, Zhao, and Ramelow, 1994
Pb C. vulgaris 5.0 Aksu and Kutsal, 1991
Cladophora prolifera 5.0 Hao, Zhao, and Ramelow, 1994
Ecklonia radiata 5.0 Matheickal and Yu, 1996
Lyngbya taylorii 3–7 Klimmek et al., 2001
Padina pavonica 2.0 Hao, Zhao, and Ramelow, 1994
Spirulina maxima 5.5 Gong et al., 2005
(Continued on next page)

S. K. Mehta and J. P. Gaur 126

TABLE 4 Optimum pH for sorption of heavy metals by algae. (Continued)
Metal Alga pH Reference

Ni Chlorella vulgaris 5.5 Mehta and Gaur, 2001a

Cladophora prolifera 5.0 Hao, Zhao, and Ramelow, 1994
Halimeda opuntia 4.0–5.0 Kuyucak and Volesky, 1989a
Lyngbya taylorii 4.0–7.0 Klimmek et al., 2001
Microcystis aeruginosa 9.2 Pradhan et al., 1998
Padina pavonica 6.0 Hao, Zhao, and Ramelow, 1994
Pilayella littoralis 5.5 Carrilho and Gilbert, 2000
Ulva lactuca 6.0 Hao, Zhao, and Ramelow, 1994
Sr Spirogyra sp. 7.0 Crist et al., 1988
Vaucheria sp. 7.0 Crist et al., 1988
Zn Cladophora prolifera 6.0 Hao, Zhao, and Ramelow, 1994
Lyngbya taylorii 3.0–7.0 Klimmek et al., 2001
Padina pavonica 6.0 Hao, Zhao, and Ramelow, 1994
Pilayella littoralis 5.5 Carrilho and Gilbert, 2000
Ulva lactuca 6.0 Hao, Zhao, and Ramelow, 1994

4. Biomass Concentration Singleton and Simmons, 1996; Fogarty et al., 1999),

The amount of a metal ion recovered from a solution
is affected by biomass concentration. Roy, Greenlaw,
and Shane (1993) showed that increasing biomass of
dried and pulverized cells of Chlorella sp. resulted in
decreased Cd binding per unit cell mass. They found
91% decrease in Cd binding by C. minutissima after a
12-fold increase in biomass concentration. Mehta and
Gaur (2001a) tested the effect of biomass concentration
on Cu and Ni sorption by C. vulgaris at different metal
concentrations. Their results show that sorption (metal
sorbed per unit biomass) of Cu and Ni was maximal
at the lowest tested biomass concentration. The above
authors have shown that increase in biomass concen-
tration from 5 mg l−1 to 1 g l−1 decreased the Langmuir
constant Qmax (maximum metal sorption capacity) for
Cu and Ni by 2.6- and 3-fold, respectively. Mehta and
Gaur (2001a) determined the effect of biomass concen-
tration on the affinity of biomass for metal binding.
They showed a decrease in affinity of C. vulgaris for Cu
and Ni with increase in biomass concentration. Gong
et al. (2005) also found a decreased Pb sorption capacity
of Spirulina maxima with increasing biomass concentra-
tion. They observed that increasing biomass concentra-
tion from 0.1 to 20 g l−1 led to a marked reduction
in Pb sorption capacity of Spirulina maxima from 121
to 21 mg g−1 . Similarly, Hamdy (2000) has noticed a
decreased sorption of Cr, Co, Ni, Cu and Cd by four
different algae with increasing biomass concentration.
Increase in biomass concentration reduces metal sorp-
tion per gram of biomass in a number of other systems
(Fourest and Roux, 1992; Brady and Duncan, 1994;
although this is generally attributed to a shift in the
sorption equilibrium. Other probable explanations for
such a relationship between biomass concentration and
sorption may be limited availability of metal, increased
electrostatic interactions, interference between binding
sites and reduced mixing at higher biomass concentra-
tions (Meikle, Gadd, and Reed, 1990; Fourest, Canal,
and Roux, 1994). Itoh, Yuasa, and Kobayashi (1975) sug-
gested that electrostatic interactions between cells could
be significant for the biomass-dependent metal sorp-
tion, with a larger quantity of metal ions being adsorbed
when distance between cells is greater. While an in-
creased biomass concentration has a negative effect on
the sorption capacity (amount of metal sorbed per unit
biomass) of a biosorbent, the total metal removed (% of
initial concentration) by a biosorbent is higher at higher
biomass concentrations. However, there is no straight-
forward relationship between biomass concentration
and metal removal. Mehta and Gaur (2001c) have
shown that increasing the biomass concentration by
100-fold enhanced Ni and Cu removal from 13 to 65%
and 10 to 85%, respectively, from solutions containing
5 mg l−1 Ni or Cu. Increased metal removal at higher
biomass concentration is simply due to greater avail-
ability of metal binding sites (Metha and Gaur, 2001c).
5. Temperature
Contrasting results have been obtained regarding
the effect of temperature on sorption of heavy met-
als by algae. Tsezos and Volesky (1981), Kuyucak and
Volesky (1989c), and Aksu and Kutsal (1991) reported

127 Use of Algae for Removing Heavy Metal Ions From Wastewater
a slight increase in cation sorption by powdered sea-
weed biomass with increase in temperature from 4 to
55◦ C. Similarly, Aksu (2002) recorded increased Ni2+
biosorption by dried biomass of Chlorella vulgaris with
enhancement of temperature from 15◦ C (48.1 mg/g)
to 45◦ C (60.2 mg/g). Increased metal sorption with in-
crease in temperature suggests that metal biosorption
by algae is an endothermic process. On the contrary,
some studies indicate exothermic nature of metal sorp-
tion by algae. For instance, Cd2+ sorption by Sargassum
sp. biomass slightly decreased with an increase in ambi-
ent temperature (Cruz et al., 2004). Some other studies
arrive at a similar conclusion (Aksu, 2001; Benguell and
Benaissa, 2002). Increased biosorption of heavy metals
with increasing temperature has been ascribed to bond
rupture that perhaps enhances the number of active sites
involved in metal sorption or higher affinity of sites
for metal. Park and Gamberoni (1997) suggested that
at higher temperatures the ions can be adsorbed more
actively on adsorption sites. Interestingly, there are re-
ports showing no effect of temperature on metal sorp-
tion (Norris and Kelly, 1979; Zhao, Hao, and Ramelow,
1994). Cossich, Tavares, and Ravagnani (2002) studied
the effect of temperature on Cr sorption by Sargassum
sp. at different pH. They have suggested that the effect
of temperature was not as pronounced as was that of
pH. Norris and Kelly (1979) and de Rome and Gadd
(1987) have shown that sorption of heavy metals is rela-
tively unaffected over a moderate range of temperature.
Likewise, Mehta, and Gaur (2001a) and Mehta, Singh,
and Gaur (2002) could not observe a pronounced effect
of temperature on extracellular binding of Cu and Ni
by Chlorella vulgaris.
Owing to dependence on metabolism, metal uptake
by live cells is considerably affected by variations in
temperature. There are reports showing altered metal
uptake by live organisms with change in temperature
regime (Skowroñski, 1986; Mehta and Gaur, 2001a;
Mehta, Singh, and Gaur, 2002), with maximum uptake
occurring at specific temperature optima.
6. Presence of Anions and Cations
Metal sorption by an algal biosorbent is also affected
by the presence of anions in the medium. Lowered ac-
cumulation of U, Co and Cu by green microalgae in
the presence of carbonate, orthophosphate, sulphate,
nitrate, EDTA and chloride ions has been observed (Rai,
Gaur, and Kumar, 1981). However, increased accumu-
lation of ionic species like nitrate and ammonium oc-
S. K. Mehta and J. P. Gaur
curred in the presence of Cu and Fe in Anabaena do-
liolum (Rai and Mallick, 1992).
The presence of other cations, including metal ions,
also significantly affects metal sorption by algae (Mehta
and Gaur, 2001a,b; Mehta, Singh and Gaur, 2002;
Mehta, Tripathi, and Gaur, 2002). Mehta and Gaur
(2001a,b) demonstrated mutual interference in sorp-
tion of Cu and Ni by Chlorella vulgaris. However, Axtell,
Sternberg, and Claussen (2003) did not find any inter-
active effect on removal of Pb and Ni by Macrospora
from a multi-metal mixture. Actual industrial wastew-
aters contain different kinds of impurities, which may
significantly affect metal biosorption (Ho and McKay,
2000). Among such impurities, light ions exist in most
industrial effluents and they greatly affect metal sorp-
tion potential of biosorbents (Lee and Volesky, 1997;
Chen and Yiacoumi, 1997; Low, Lee, and Liew, 2000).
However, Adhiya et al. (2002) did not find inhibitory
effect of Ca2+ and K+ on Cd sorption by lyophilized
Chlamydomonas reingardtii. Reduced metal uptake in the
presence of light metals is attributed to competition for
cellular binding sites, or precipitation or complexation
by carbonates, bicarbonates or hydroxides of Ca and
Mg (Rai, Gaur, and Kumar, 1981). Other compounds
that could be considered as impurities in metal removal
process are surfactants and some chelating agents. The
nature of impurities differs depending on the type of
effluent to be treated. High concentrations of salts
like NaCl in solution also decrease the rate of metal
sorption by algae (Cho et al., 1994). Garnham, Codd,
and Gadd (1993a) and Corder and Reeves (1994) have
shown that Na decreases the accumulation of Ni, Co
and Cs by some algae and cyanobacteria. Yun, Niu,
and Volesky (2001) also noted suppression of sorption
of Cr (VI) and Va by Na and Cl. High concentrations
of monovalent cations Na+ and K+ , increase the ionic
strength of a wastewater. The increased ionic strength of
a wastewater often decreases metal biosorption capacity
of the biomass (Greene, McPherson, and Darnall, 1987;
Ramelow, Fralick, and Zhao, 1992). The inhibitory ef-
fect of Na is more pronounced with weakly bound
metals such as Zn or Ni. It is important to note that
Na+ and K+ , being monovalent cations, do not com-
pete directly with covalent binding of heavy metals by
the biosorbent. Whereas most of the studies report in-
hibitory effect of light metal ions on sorption of heavy
metals by biosorbent, a few of them show no effect.
For example, Pawlik and Skowroñski (1994) could not
observe any change in Cd transport in Synechocystis in
128
the presence of K. Likewise, Cs sorption capacity of
Padina australis did not change in the presence of Na or
K in the solution (Jalali-Rad et al., 2004).
7. Other Factors
Nutrient level, growth rate and illumination greatly
influence metal sorption by living algae. Hall, Healey,
and Robinson (1989) noticed enhanced uptake of
Cu by Chlorella at limiting PO−3 4 concentration. Up-
take of Cu, Cd and Zn by Aphanocapsa increased
with increase in NO− 3 concentration in the medium
(Subramanian, Sivasubramanian, and Gowrinathan,
1994). Algal growth increases with increasing light in-
tensity to saturation level, but the influence of light
intensity on metal sorption remains largely unknown.
There are reports showing inhibition of metal sorp-
tion in the dark (Garnham, Codd, and Gadd, 1992;
Pawlik and Skowroñski, 1994). However, Subramanian,
Sivasubramanian, and Gowrinathan (1994) have shown
higher Zn uptake in the dark. Metal sorption is also in-
fluenced by the growth phase of algal culture. The ad-
sorption of Ni on the surface of C. vulgaris was higher
for cultures in the stationary and decline phase than
those in the exponential phase (Mehta, Tripathi, and
Gaur, 2000). This may have resulted due to better expo-
sure of metal binding sites or creation of additional sites
on the cell surface during these phases (Mehta, Tripathi,
and Gaur, 2000). Variations in growth conditions pos-
sibly bring about changes in composition of the cell
surface thereby affecting metal biosorption characteris-
tics of the biomass.
B. Live vs. Inactivated Biomass
for Metal Sorption
A majority of studies on metal accumulation by al-
gae have been performed with living organisms for
environmental, toxicological and pharmaceutical pur-
poses rather than with industrial application in mind.
Of late, the attention has shifted to non-living algae
and other microorganisms for metal removal and/or
recovery (Volesky and Holan, 1995; Aksu, 1998). In
comparison to live cells, the metal sorption capacity
of dead cells may be greater, equivalent or less (Özer,
Özer, and Dursun, 2000). The process of using dead
cells can be of great interest, because of the large vari-
ety and low cost of these biological materials. In gen-
eral, inactivated cells display greater metal binding ca-
pacity than live cells. They do not require a nutrient
129
supply, and therefore can be used for multiple sorption-
desorption cycles. However, vacuum drying decreased
uranium sorption by Scenedesmus obliquus from 13 to
0.6 mg g−1 dry weight (Zhang et al., 1997). Decreased
metal uptake after vacuum drying of the biomass may
be due to disappearance or closure of the effective
sites, capillaries or caves for captivating metal ions on
the cell wall as water evaporates from the cells un-
der vacuum condition (Zhang et al., 1997). Conversely,
it is widely reported that freeze-dried biomass has a
higher metal sorption potential than the live biomass
(Winter, Winter, and Pohl, 1994; Bengtsson et al., 1995).
Neide, Carrihlo, and Gilbert (2000) observed no change
in metal sorbing potential of freeze- and oven-dried
biomass, when compared with live biomass. Burdin and
Bird (1994) studied metal sorption by living as well
as lyophilized Gracilaria tikvahiae, Gelidium pusillum,
Agardhiella subulata and Chondrus crispus. They found
greater accumulation of Pb in living thalli of all the
four species compared to lyophilized thalli. However,
they noticed greater accumulation of Ni, Cu and Zn
by lyophilized than living thalli of Gracilaria, Gelidium
and Chondrus. The above authors found no difference
between lyophilized and living thalli of the selected sea-
weeds for accumulating Cd. While contrasting effect of
lyophilization has been found, Adhiya et al. (2002) re-
ported that lyophilized cells and living cells of Chlamy-
domonas reinhardtti showed similar ATR-FTIR spectra.
It suggests that lyophilization does not change chemi-
cal composition of the cell surface, including cell wall.
Metal sorption capacity of dead cells depends on their
pretreatment and any subsequent change in the struc-
ture of their cell wall (Somers, 1963; Duddridge and
Wainright, 1980). The method of inactivation of cells
also influences their metal sorption capacity. Whereas
heat killing of biomass may enhance sorption of met-
als, chemical killing may at times decrease the metal
sorption capacity of the biomass (Tobin, Cooper, and
Neufeld, 1990). Pretreatment of Chlorella vulgaris with
dilute HCl considerably increased Ni and Cu sorption
from aqueous solution (Mehta and Gaur, 2001a; Mehta,
Tripathi, and Gaur, 2002).
Whereas the use of inactivated biomass has been
preferred, some disadvantages also deserve mention.
Dead cells cannot be used where biological alteration
in valency of a metal is sought. Moreover, degrada-
tion of organometallic species is not possible with dead
biomass. Another important drawback associated with
dead biomass is that there is no scope for biosorption
Use of Algae for Removing Heavy Metal Ions From Wastewater
improvement through mutant isolation. The use of live
biomass has several demerits; potential for desorptive
metal recovery is limited since metal may be intracellu-
larly bound, and metabolic extracellular products may
form complexes with metals to retain them in solution.
C. Metal Sorption by
Pretreated Biomass
Although many biological materials bind heavy met-
als, only those with a sufficiently high metal binding
capacity are suitable in full-scale biosorption process.
Various treatments can be given to increase the metal
sorption capacity of the biomass (see Table 5). In com-
parison to fungal and other biosorbents, fewer searches
have been conducted for suitable agents and conditions
to increase the inherent metal sorption ability of algal
biosorbents. Several studies show that CaCl2 pretreat-
ment is the most suitable and economic method for ac-
tivation of algal biomass. CaCl2 pretreatment increased
Pb sorption capacity of Spirulina maxima by 84–92%.
Feng and Aldrich (2004) have also used CaCl2 for ac-
tivation of Ecklonia maxima for Cd, Cu and Pb sorp-
tion. Yu and Kaewsarn (1999) have shown an increased
Cu sorption following CaCl2 treatment of the marine
alga Durvillaea potatorum. Kaewsarn and Yu (2001) pre-
treated Padina sp. with 0.2 M CaCl2 , and noted that
the pretreated biomass bound 0.5 mmol g−1 Cd, and
in a column removed 98% of Cd within 35 min. Gong
et al. (2005) observed 84–92% increase in Pb sorption
capacity of Spirulina maxima after CaCl2 pretreatment
of the biomass. Next to CaCl2 , mineral acids (e.g., HCl
and HNO3 ) have been found highly effective in in-
creasing metal sorption potential of some algae. On
the basis of electron micrograph of pretreated Durvil-
laea potatorum, Yu, Kaewsarn and Duong (2000) have
reported that CaCl2 pretreatment followed by thermal
treatment provides most uniform cylinder or fiber-like
shapes for the biosorbent. Mehta and Gaur (2001a) used
several chemicals to enhance metal removal capacity of
Chlorella vulgaris. Pretreatment with dilute HCl consid-
erably increased sorption of Cu and Ni by C.vulgaris.
Other agents like methanol, acetic acid, NaOH and hot
water nevertheless decreased metal sorption. HCl pre-
treatment of biomass was effective in enhancing metal
sorption from single as well as binary metal solutions
(Mehta, Tripathi, and Gaur, 2002). Kalyani, Rao, and
Krishniah (2004) demonstrated enhanced Ni sorption
capacity of Sargassum sp. from 181.2 to 250 mg g−1
S. K. Mehta and J. P. Gaur
following acid pretreatment. Zhao, Hao, and Ramelow
(1994) studied effects of different pretreatments on sub-
sequent sorption of several metal ions from solution by
six strains of seaweeds. All the pretreatments increased
the ability of the biomass to bind metals (Pb, Cu, Zn,
Cd, Cr, Mn, Ni, Co, etc.). Zhang et al. (1997) showed
decreased uranium sorption by Scenedesmus obliquus fol-
lowing pretreatment with dilute HCl, NaOH, NaCl and
ethanol. They assumed that functional groups taking
part in uranium sorption were on the cell surface and
also that pretreatment of live cells by alcohol, NaOH,
NaCl, or even water perhaps caused contraction of cells,
thereby making sites less accessible to metal ions. De-
crease in U uptake due to HCl pretreatment of the
biomass was probably due to penetration of protons
(with smaller radius than uranium) somewhere into the
surface and this probably increased rigidity of macro-
molecules that might otherwise have been involved in
metal binding (Zhang et al., 1997). Mehta, Tripathi, and
Gaur (2002) demonstrated that acid pretreatment not
only increased metal sorption but also was able to allevi-
ate the inhibitory effect of other metal ions on sorption
of the metal of interest.
Xanthanation of biomass has been recently intro-
duced as a novel approach to increase biosorption of
Pb by algal biomass. Xanthanation of biomass increased
sulfur content of the Undaria pinnatifida biomass from
0.15% to 13.7%, and xanthanated biomass sorbed three
times more Pb than the native biomass (Kim et al., 1999).
Kim et al. (1999) carried out xanthanation of U. pinnat-
ifida in two steps. In the first, hardening of cell wall
was carried out by cross-linking using epichlorohyrin.
In the second step, xanthanate group was introduced
by chemical reaction of the biomass with carbon disul-
fide. However, Kim et al. (1999) have shown that acid
treatment decreased sulfur content of the xanthanated
biomass indicating its instability in acidic condition.
Due to instability of xanthate in acidic conditions,
HCl or HNO3 should not be used for desorption of
bound metal. Klimmek et al. (2001) described a new
method for increasing metal sorption capacity of algal
biomass. They introduced phosphate groups in Lyn-
gbya taylori in order to increase the metal removal
capacity of the biomass. The additional insertion of
phosphate groups into the cell wall was achieved by
esterification of hydroxyl groups of the polysaccharides
using phosphoric acid in a urea solution. They showed
that phosphorylation increased phosphate content of
the biomass from 0.6 to 4.4 mmol g−1 of biomass,
130
TABLE 5 Effect of biomass pretreatment on its metal sorption efficiency.
% increase
(+) or
decrease (−)
in metal
Biomass Pretreatment Metal sorption Reference
Ascophyllum nodosum HCl (0.01 N) Co −74 Kuyucak and Volesky, 1989a
KSCN (0.1 N) Co −19 Kuyucak and Volesky, 1989a
Methanol-chloroform, 1:3, 30 h Co −3 Kuyucak and Volesky, 1989a
NaOH (0.01 N) Co −68 Kuyucak and Volesky, 1989a
Warm water (60◦ C), 1 h Co −17 Kuyucak and Volesky, 1989a
Chlorella vulgaris Acid Au +54 Ting, Teo, and Soh, 1995
Alkali Au +25 Ting, Teo, and Soh, 1995
CH3 COOH (0.1 mM) Cu −18 Mehta and Gaur, 2001a
CH3 COOH (0.1 mM) Ni +16 Mehta and Gaur, 2001a
Formaldehyde Au +52 Ting, Teo, and Soh, 1995
HCl (0.1 mM) Cu +20 Mehta and Gaur, 2001a
HCl (0.1 mM) Ni +78 Mehta and Gaur, 2001a
Heat Au +14 Ting, Teo, and Soh, 1995
HNO3 (0.1 mM) Cu +13 Mehta and Gaur, 2001a
HNO3 (0.1 mM) Ni +54 Mehta and Gaur, 2001a
Hot water Cu −45 Mehta and Gaur, 2001a
Hot water Ni −53 Mehta and Gaur, 2001a
Methanol (90%) Cu −20 Mehta and Gaur, 2001a
Methanol (90%) Ni −39 Mehta and Gaur, 2001a
NaOH (0.1 mM) Cu +48 Mehta and Gaur, 2001a
NaOH (0.1 mM) Ni −1 Mehta and Gaur, 2001a
De-alginated sea weed Esterification Cd −82 Romero-Gonzalez, Williams, and
Gardiner, 2001
Laminaria japonica CaCl2 Cd +30.0 Yin, et al., 2001
Lyngbya taylorii Phosphorylation Cd +581 Klimmek et al., 2001
Ni +329 Klimmek et al., 2001
Pb +109 Klimmek et al., 2001
Zn +430 Klimmek et al., 2001
Oscillatoria anguistissima 0.01 M HCl Zn −48 Ahuja et al., 1999
0.01 N NaOH Zn −6 Ahuja et al., 1999
Drying at 80◦ C for 48 h Zn +29 Ahuja et al., 1999
Ulva lactuca 0.05 M ammonium acetate, 15 min. Cd +36 Hao, Zhao, and Ramelow, 1994
Cu +85 Hao, Zhao, and Ramelow, 1994
Pb +8 Hao, Zhao, and Ramelow, 1994
Zn +37 Hao, Zhao, and Ramelow, 1994
1 M KOH, 15 min Cd +41 Hao, Zhao, and Ramelow, 1994
Cu +85 Hao, Zhao, and Ramelow, 1994
Pb +13 Hao, Zhao, and Ramelow, 1994
Zn +87 Hao, Zhao, and Ramelow, 1994
1 M KOH Cd +95 Hao, Zhao, and Ramelow, 1994
Cu +120 Hao, Zhao, and Ramelow, 1994
Pb +21 Hao, Zhao, and Ramelow, 1994
Zn +90 Hao, Zhao, and Ramelow, 1994
1 M Ammonium acetate Cd +40 Hao, Zhao, and Ramelow, 1994
Cu +125 Hao, Zhao, and Ramelow, 1994
Pb +19 Hao, Zhao, and Ramelow, 1994
Zn +30 Hao, Zhao, and Ramelow, 1994
Triton X-100 Cd +6 Hao, Zhao, and Ramelow, 1994
Cu +13 Hao, Zhao, and Ramelow, 1994
Pb +11 Hao, Zhao, and Ramelow, 1994
Zn +6 Hao, Zhao, and Ramelow, 1994
Sargassum fluitans L-cysteine Au +250 Niu and Volesky, 2000
Spirulina maxima 0.2 M CaCl2 , 24 h, biomass washed with Pb +10 Gong et al., 2005
deionized water

131 Use of Algae for Removing Heavy Metal Ions From Wastewater
thereby enhancing Cd2+ sorption capacity of L. taylori
sevenfold.
D. Use of Immobilized Algae
in Sorption of Metals
Algal biomass cannot be used directly in a standard
sorption process. It is generally of small particle size and
low strength and density, which can limit the choice of
a suitable reactor and make biomass or effluent separa-
tion difficult (Tsezos, 1986). The use of native or even
powdered algal biomass is not desirable as it may clog
the column. However, biomass immobilized within or
on an inert matrix has the inherent advantage that high
flow rate can be achieved with minimal clogging of the
column, and also that the control of size of sorbent par-
ticle and high biomass loading of the column reactor
are feasible (Shumate et al., 1980; Tsezos, 1986; Yakubu
and Dudeney, 1986). Efforts have been made to test the
efficacy of immobilized algae and cyanobacteria for the
removal of heavy metals from aqueous solution. Immo-
bilization is a general term that describes many different
forms of cell attachment or entrapment (Lopez, Lazora,
and Marques, 1997). Various techniques, such as floc-
culation, adsorption on surfaces, covalent binding to
carriers, crosslinking of cells, encapsulation in polymer
gel and entrapment in polymeric matrix, are used for
cell or biomass immobilization.
For immobilization of biomass, the supporting ma-
terial is either natural, including agar, alginate and
carrageenan, or synthetic, such as silica gel, polyacry-
lamide and polyurethanes. Natural polymers are bet-
ter than synthetic polymers due to the latter’s toxi-
city to biomass. Alginate has been most extensively
used for immobilization of algal as well as other kinds
of biomass. Alginate is extracted from algae as water-
soluble sodium salt. When calcium replaces sodium,
ionic cross-linking between carboxylic acid groups oc-
curs giving a gelatinous substance. It may be noted that
alginate beads are vulnerable to low as well as high pH,
and are stable when the pH of the external medium is
between 5–9. At pH below 3 and above 9, there is a
considerable loss of alginate. At pH 11, alginate beads
most often break. Treatment of highly acidic effluent
by alginate-immobilized biomass may result in shrink-
age of biomass-loaded beads. Furthermore, when algi-
nate is used as immobilization support one cannot use
acid or alkali during the desorption process. Leakage
of biomass is more likely when alginate is used as ma-
S. K. Mehta and J. P. Gaur
trix. Biomass leakage from alginate beads occurs when
orthophosphate or Na+ contacts with the alginate im-
mobilized system. This system is therefore not good
for the treatment of wastewaters that contain high lev-
els of Na+ or orthophosphate. Among synthetic poly-
mers, polyacrylamide has been most extensively used
(Darnall et al., 1986; Robinson and Wilkinson, 1994).
Polyacrylamide immobilization is not prone to dam-
age by cation replacement or chelation unlike the com-
monly used calcium alginate system; however, high cost
and toxicity to living cells restricts its application in im-
mobilization process. It has been shown that mechan-
ical strength of organic gel materials, such as alginate
and polyacrylamide, is not high enough to permit col-
umn operation on a large scale because they may be
crushed due to column bed pressure (Hu and Reeves,
1997). Silica gel has also been used for entrapment of
algal cells or biomass (Rangsayatorn et al., 2004). Silica
gel is prepared by decreasing the pH of alkali silicate to
less than 10. The solubility of the silica is then reduced
to form the gel. As silica begins to gel, cells in silica are
trapped in porous gel that is three-dimensional SiO2
network, of which 60% is water.
Akhtar, Iqbal, and Iqbal (2003) developed a new
biosorbent by immobilizing Chlorella sorokiniana within
luffa sponge discs and showed that microalgal-luffa
sponge disc system has good biosorption properties
with respect to Ni. Microalgal-luffa sponge immobi-
lized discs removed Ni very rapidly, with 97% of equi-
librium loading being reached in 5 min. The regener-
ated microalgal-luffa sponge immobilized disc retained
92.9% of the initial binding capacity for Ni up to five
cycles of reuse in fixed bed column reactor.
Immobilization generally tends to increase metal ac-
cumulation by biomass (Darnall et al., 1986; Aksu, 1998;
Guo et al., 2000). Immobilized cells accumulate more
metals than free cells due inter alia to (i) enhanced
photosynthetic capacity (Khummongkol, Canterford,
and Freyer, 1982), and (ii) increased cell wall perme-
ability (Brouers et al., 1989). Immobilization of living
biomass also provides protection to cells from metal
toxicity (Bozeman, Koopman, and Bitton, 1989). On
the contrary, some reports show a higher metal sorb-
ing efficiency of free cells compared to immobilized
cells (Wong and Pak, 1992). For example Rangsayatorn
et al. (2004) showed that free cells of Spirulina platensis
accumulated 98 mg g−1 while alginate and silica im-
mobilized cells accumulated 71 and 37 mg g−1 Cd, re-
spectively. Bag, Lale, and Turker (1999) obtained similar
132
results. Change in the structure of cell wall during
immobilization process is perhaps responsible for de-
creased metal sorption capacity of the immobilized
biomass (Rangsayatorn et al. (2004)). It is also probable
that part of cell surface might be shielded by the gel ma-
trix making sites for metal binding unavailable. Size of
immobilized bead is a crucial factor for use of immobi-
lized biomass in biosorption process. Beads should not
be extremely small as the column filled with them might
quickly choke. Due to low pressure drop in column and
poor intraparticle mass transfer, bigger beads are also
not recommended (Volesky, 2001). It is recommended
that beads should be in the size range between 0.7 and
1.5 mm, corresponding to the size of commercial resins
meant for removing metal ions (Volesky, 2001).
Since immobilization of a biosorbent is costly per se
(Sag and Kutsal, 1998), a few researchers have used dried
biomass or converted it into granules by certain chem-
ical treatments (Holan, Volesky, and Prasetyo, 1993;
Volesky and Prasetyo, 1994). A few reports are avail-
able on cross-linking of cells with various chemical com-
pounds. Several cross-linking materials could be found
in literature (Holan, Volesky, and Prasetyo, 1993): alde-
hydes, polysaccharides, sulfones, vinylketones, epox-
ides, etc. Each such material has a specific group ca-
pable of cross-linking with the cell wall of the biomass.
Formaldehyde forms a chemical cross-linking between
adjacent hydroxyl groups of sugars of the cell wall. Glu-
taraldehyde cross-links with amino groups of cell wall
due to the presence of prolonged carbon chain. In cross
linking with polyethylene imine, biomass is first embed-
ded in polyethylene imine, and free amino groups of
the matrix are further cross-linked with glutaraldehyde,
which gives further stability to the immobilized system.
Glutaraldehyde and polyethylene imine treatments of
biomass introduce amino and some other chemical
groups thereby altering metal sorption (Leusch, Holan,
and Volesky, 1995). However, chemical treatment for
cross-linking of biomass may decrease metal sorption
(Jalai-Rad et al., 2004).
The immobilized system should be stable, and light
should be able to pass through easily if live algal biomass
is used. It should not discharge the biomass during
treatment. For an immobilized cell system to be effec-
tive, the process of immobilization must not cause irre-
versible, structural, physiological and metabolic dam-
age to the cell. Immobilized biomass in the form of
beads should not expand or swell during metal removal.
Some algae exist naturally in an immobilized state ei-
133
ther as films on surface or entrapped within gel or slime
of their own synthesis (Brouers et al., 1989). Such algae
can be exploited for removing metals from wastewaters.
Although immobilized and granulated algae have great
potential in removing toxic metal ions from wastewa-
ters, there is a need to develop an economically feasible
method for producing and using them.
E. Metal Sorption From Binary
and Multi-Metal Solutions
Many industrial wastewaters contain high levels of
more than one metal. For instance, the wastewater aris-
ing from fur cleaning and dyeing industry has been
shown to contain 7.04, 20.14, 0.17, 1.73 and 0.12 mg
l−1 of Cu, Cr, Ni, Zn and Cd, respectively (Klein et al.,
1974). Similarly, Cr and Cu are frequently encountered
together in industrial wastewaters, e.g., from mining,
metal cleaning, plating, electroplating, metal process-
ing, dyeing and petroleum industries. In metal clean-
ing, plating and metal processing industries, Cu and Cr
concentrations may approach 100–120 mg l−1 and 10–
270 g l−1 , respectively (Sag and Kutsal, 1996). Effluent
coming out from mining operations have Cu, Zn, Pb,
Cr, As and Se together (Volesky, 2001). Cr, Ni, Cd and
Zn are reported to occur together in effluents gener-
ated during electroplating operations (Volesky, 2001).
Most of the industrial effluents have high concentra-
tions of Al along with other metal ions. Although Al
is not a major environmental problem, its ubiquitous
presence in solution interferes with sorption of many
other metals (Lee et al., 2004). While the accumulation
of single species of heavy metal ions by algal biomass
has been extensively studied, very little attention has
been given to the study of multi-metal system. The pres-
ence of a multiplicity of metals leads to interactive ef-
fects on physiological and biochemical processes, e.g.,
growth, metal uptake, etc., of various organisms. (Ting,
Lawson, and Prince, 1991). Studies on multi-metal sys-
tems have shown competitive interactions amongst
metals for binding onto sorption sites. Cd and Fe in-
terfere sorption of each other by Sargassum sp from the
binary metal solution (Figueira, Volesky, and Ciminelli,
1997). Metals differ with regard to their ability to inter-
fere with sorption of other metals. For example, 1.5 mM
Fe caused 24% reduction in Cd biosorption, but at a
similar concentration Cd caused up to 45% reduction
in sorption of Fe by Sargassum from a binary metal solu-
tion (Figueira, Volesky, and Ciminelli, 1997). Lee et al.
Use of Algae for Removing Heavy Metal Ions From Wastewater
(2004) reported that 1 mM concentrations of Pb and
Al at pH 4.5 mutually reduced Al and Pb uptake by
Laminaria japonica to 22.3 and 83%, respectively. Most
of the time, total adsorption capacity of biosorbent is
lower for binary metal system than the single compo-
nent due to inhibitory effect of one metal on binding
of other metal. Mehta, Tripathi, and Gaur (2000) have
shown mutual interference in sorption of Cu and Ni by
Chlorella vulgaris from the binary metal solution. Inhibi-
tion in Ni sorption due to Cu was stronger than inhibi-
tion in Cu sorption due to Ni. Despite the fact that one
metal ion present in a solution may decrease sorption
of the other metal ion and vice versa, overall metal sorp-
tion does not necessarily decrease (Figueira, Volesky,
and Ciminelli, 1997). Apiratikul et al. (2004) found that
overall adsorption capacity of Caulerpa lentifollifera for
Cd and Cu was higher than adsorption of Cd alone,
but was lower than adsorption of Cu in single compo-
nent system. This indicates that binding sites for Cd
are not the same as that for Cu. According to them,
Caulerpa lentifollifera has common binding sites for Cu
and Pb, and Cu and Pb showed competitive inhibition
FIGURE 2 Copper and Pb sorption by Spirogyra granules from
for sorption of each other in the binary system. Further, binary metal solution containing different concentrations of Cu
competitive interaction between two metals depends on and Pb. The pH of the solution was 4.5 ± 0.2 and the biomass
the concentration of algal biomass in the solution. Terry concentration was 1 g l−1 . The contact time of biomass with metal
solution was 30 min (based on unpublished data of Alpana Singh).
and Stone (2002) determined the effect of biomass con-
centration on sorption of Cd and Cu from binary metal senting metal sorption characteristics of a biosorbent in
solution by Scenedesmus abundans. They have shown that a multi-metal system. Figure 2 shows the sorption of
at the lowest tested biomass concentration, competitive Cu and Pb by Spirogyra from a binary metal solution,
effect was observed at Cu and Cd concentrations above and makes it amply clear that Pb inhibited Cu sorption
4 mg l−1 each. At the highest algal concentration con- more intensely than Cu inhibited Pb sorption.
sidered, no competitive effect occurred at Cu and Cd
concentration in the range 1–7 mg l−1 . Al presents an
F. Regeneration and Reuse
interesting example of interference in heavy metal sorp-
tion by algae. Lee et al. (2004) found that Al decreases
of Biomass
sorption of Pb, Cu, Cd and Zn by 78, 80, 86 and 89%, re- In order to make the biosorption-based process
spectively in Laminaria japonica. However, Al caused no feasible for industrial application, provision must be
significant decrease in Cr (III) sorption. They suggested made to regenerate the biomass for repeated use. Metal
that being trivalent Al could not compete with divalent sorbed on biomass can be desorbed by a suitable eluant
metal ions for binding on surface sites. Although metal or desorbing solution, and thus biomass can be used
sorption ability of the biomass decreases in multi-metal in multiple sorption-desorption cycles. One of the
system, multi-metal solutions can be remediated to the most common methods for desorption of heavy metals
same level as single metal solutions at high biomass con- from the biomass could be the swing in pH. Lowering
centrations (Mehta, Tripathi, and Gaur, 2002; Terry and the pH of the metal-loaded biomass suspension causes
Stone, 2002). When more than one metal are present displacement of heavy metal cations by protons from
in sorption system the evaluation of biosorption re- the binding sites. In an experiment with Chlorella
sults, their interpretations and representation become minutissima, Roy, Greenlaw, and Shane (1993) found
much more complicated. Volesky and Holan (1995) and about 90% desorption of Pb and Zn by lowering the
Mehta and Gaur (2001c) advocated 3D plots for repre- pH of the suspension to 1.55. However, the biomass

S. K. Mehta and J. P. Gaur 134

may become damaged at extremely low pH. Organic
and mineral acids, bases, salts and metal chelators have
been tested for their metal desorbing ability (Kuyucak
and Volesky, 1989b; Hu and Reeves, 1997; Esteves,
Valdman, and Leite, 2000). The tested desorbing
agents include hydrochloric acid, sodium hydroxide,
acetic acid, citric acid, mono- and dibasic ammonium
phosphate, ammonium sulfate, ammonium chloride,
sodium carbonate, calcium chloride, EDTA, etc. Zhou,
Huang, and Lin (1998) could recover up to 99.5% of
Cu and Cd from Laminaria japonica and Sargassum
kjellmarianum by HCl and EDTA. Vijayaraghavan
et al. (2005) tested the efficiency of HCl, H2 SO4 ,
HNO3 and CaCl2 for desorbing Cu bound on Ulva
reticulata. CaCl2 (0.1 M in HCl or HNO3 ) gave
the best results. Jalai-Rad et al. (2004) found almost
complete desorption of Cs from Sargassum glauescens
and Cystoseria indica by 0.5 N NaOH or 1 M KOH. But
Cs sorption capacity of biosorbents decreased by up
to 51% after 9 sorption/desorption cycles. Chojnacka,
Chojnacki and Górecka (2005) studied desorption
of Cr3+ , Cd2+ and Cu2+ from Spirulina sp. using
0.1 M EDTA, 0.1 M HNO3 and deionized water. They
found that the most efficient desorbent was nitric
acid (0.1 M) which removed almost all the metal ions
(98%) bound with the biomass and did not cause
the loss of biosorption capacity. Although nitric acid
(1 M) desorbed all the metal ions simultaneously, it
damaged the biomass and caused a significant decline
of biosorption capacity. Desorption efficiency of 0.1 M
EDTA ranged between 53 to 63%. In a column study
with Sagassum kjellmanianum, Zhou, Huang, and Lin
(1998) observed dramatic increase in Cu concentration
in desorbing solutions from 0 to 4991 mg l−1 and
4659 mg l−1 , after elution with 20 ml of HCl (0.11 N)
and EDTA (0.1 N), respectively. Approximately 98%
of the sorbed Cu was desorbed with 50 ml of elutant.
The regenerated column could bind Cu with the
same efficiency, thus suggesting that such biosorption
systems can be fully regenerated and reused. Karna et al.
(1999) studied the reusability of alkali-extracted (5%
KOH) marine cyanobacterium Phormidium valderianum
over up to six sorption/desorption cycles. They tested
several agents for desorption of Cd and Co from
biomass and found HCl and EDTA the most efficient;
other desorbing agents were found to have lower
efficiency in the following order: H2 SO4 > NH4 Cl>
CaCl2 >Na2 SO4 > KSCN>KCl>NH4 OH>NaHCO3 .
They further reported that more than 80% of Cd bind-
135 Use of Algae for Removing Heavy Metal Ions From Wastewater
ing capability was retained up to three cycles of
reuse, after which it decreased to 40–45%. Co binding
capacity of the biomass was rapidly lost after HCl
desorption, and 70% of the initial binding capacity
was lost by the sixth cycle. Rangsayatorn et al. (2004)
tested the reusability of Spirulina platensis TISTR
8217 immobilized in alginate and silica gel in five
cycles of Cd adsorption and desorption using 0.1 M
HCl as the desorbing agent. Their results show a
significant loss (ca. 26%) in Cd adsorption by the
biomass after the first cycle; however, Cd adsorption
was almost constant after the second cycle. Hashim,
Tan and Chu (2000) also found 50% reduction in
Cu sorption capacity of Sargassum bacularia after
desorption wash with HCl. Likewise, a 74% decreased
Co uptake capacity of the HCl-eluted Ascophyllum
nodosum was observed during cobalt elution (Kuyucak
and Volesky, 1989a). Although HCl has a high capacity
to desorb metals, several studies show that it decreases
metal sorption ability of biosorbent in successive
cycles due probably to damage to metal binding sites
and hydrolysis of polysaccharides occurring on the
surface of the biomass (Chu et al., 1997). Therefore,
the use of HCl for regeneration of algal biomass
cannot be recommended. Although EDTA is a strong
metal chelator, it is not safe for the environment.
Furthermore, EDTA could not be used for desorption
wash of alginate immobilized biomass because EDTA
dissolves alginate. The elutant for desorbing metals
from biomass has to be carefully selected. It should
be capable of desorbing bound metal quickly and
almost completely, and must also function as an
electrolyte in order to recover the metal by electrolytic
process (Butter et al., 1998). Therefore, CaCl2 and
NH4 Cl seem to be suitable desorbing agents. Another
important point is that the desorbing agent should not
decrease the metal sorption capacity of biomass during
successive cycles of metal sorption. Acid treatment
cannot be used for removing anionic forms of metals
such as Cr (VI); NaOH has been found particularly
effective in desorbing hexavalent Cr from biosorbents
(Lee et al., 2000). In order to design and optimize a
biosorption process for industrial application, it is
extremely important to elucidate metal sorption and
desorption behavior of biomass. Table 6 summarizes
metal elution efficiency of different desorbing agents.
Although an absolute comparison of different desorb-
ing agents cannot be made due to varying experimental
conditions used by different researchers, it is reasonable
TABLE 6 Metal elution efficiency of different desorbing agents.
Desorption of
Desorbent Metal Biosorbent sorbed metal (%) Reference

CaCl2 , 0.05 mM Cd Sargassum sp. 42 Esteves, Valdman, and Leite, 2000

CaCl2 , 0.05 mM Zn Sargassum sp. 48 Esteves, Valdman, and Leite, 2000
CaCl2 , 0.1 M Cd Phormidium valderianum 11 Karna et al., 1999
CaCl2 , 0.1 M Co P. valderianum 40 Karna et al., 1999
CaCl2 /HCl, 1:3, pH 2.5 Co Ascophyllum. nodosum 96 Kuyucak and Volesky, 1989d
Citric acid Pb Spirulina maxima 75 Gong et al., 2005
EDTA, 0.1 M Cd Phormidium valderianum 100 Karna et al., 1999
EDTA, 0.1 M Co P. valderianum 100 Karna et al., 1999
EDTA Cu Sargassum kjellmanianum 98 Zhou et al., 1998
EDTA, 0.1 M Pb Spirulina maxima 91 Gong et al., 2005
EDTA, pH 2.5 Co Ascophyllum nodosum 60 Kuyucak and Volesky, 1989d
H2 SO4 , 0.1 M Cd Phormidium valderianum 47 Karna et al., 1999
H2 SO4 , 0.1 M Co P. valderianum 75 Karna et al., 1999
HCl, 0.1 M Zn Sargassum sp. 60 Esteves, Valdman, and Leite, 2000
HCl, 0.1 M Cd Phormidium valderianum 43 Karna et al., 1999
HCl, 0.1 M Co P. valderianum 78 Karna et al., 1999
HCl, 0.11 N Cu Sargassum kjellmanianum 98 Zhou et al., 1998
HCl, 0.5 M Cd Sargassum sp. 100 Esteves, Valdman, and Leite, 2000
HCl, 1 N Cr Pachymeniopsis 0.25 Lee et al., 2000
HCl, pH 1.55 Pb Chlorella minutissima 90.2 Roy et al., 1993
HCl, pH 1.55 Zn C. minutissima 91.5 Roy et al., 1993
HCl, pH 1.55 Cd C. minutissima 1.2 Roy et al., 1993
HCl, pH 1.55 Ni C. minutissima 0.3 Roy et al., 1993
HCl, pH 2.7 Co Ascophyllum nodosum 91 Kuyucak and Volesky, 1989d
HCl, 0.1 M Pb Spirulina maxima 85 Gong et al., 2005
HCl:EDTA, (1:1) Cd Padina pavonica 90 Ofer, Yerachmiet, and Shmuel, 2003
HCl:EDTA, (1:1) Ni Padina pavonica 90 Ofer, Yerachmiet, and Shmuel, 2003
HNO3 , 0.1 M Pb S. maxima 92 Gong et al., 2005
KHCO3 , pH 6 Co A. nodosum 57 Kuyucak and Volesky, 1989d
KSCN, 0.1 M Cd Phormidium valderianum 3 Karna et al., 1999
KSCN, 0.1 M Co P. valderianum 13 Karna et al., 1999
KSCN, pH 2.5 Co Ascophyllum. nodosum 98 Kuyucak and Volesky, 1989d
NaHCO3 , 0.1 M Cd Phormidium valderianum 6.4 Karna et al., 1999
NaOH, 1 N Cr Pachymeniopsis 17 Lee et al., 2000
NaOH, 0.1 M Cd Sargassum sp. <6 Esteves, Valdman, and Leite, 2000
NaOH, 0.1 M Zn Sargassum sp. <6 Esteves, Valdman, and Leite, 2000
NH4 Cl, 0.1 M Co Phormidium valderianum 30 Karna et al., 1999
NH4 Cl, 0.1 M Cd P. valderianum 20 Karna et al., 1999
NH4 Cl, pH 2.8 Co Ascophyllum nodosum 78 Kuyucak and Volesky, 1989d
NH4 OH, 0.1 M Cd Phormidium valderianum 2.2 Karna et al., 1999
NH4 OH, 0.1 M Co P. valderianum 11 Karna et al., 1999

to conclude that CaCl2 and mineral acids like HCl are
highly efficient in desorption of metal cations from
algal biosorbents. Table 6 also shows that efficiency
of a desorbing agent also depends on algal species.
For example, HCl desorbed 100% Cd from Sargassum
sp. but only 1.2% Cd from Chlorella minutissima. This
suggests differences in the mechanism of metal sorp-
tion in brown and green algae that are known to dif-
fer in cell wall composition. Therefore, it is necessary
to screen different desorbing agents for maximum re-
covery of metal ions from a wastewater. During selec-
tion of desorbing agent evaluation should be made for
both desorption efficiency as well as preservation of
biosorption capacity of biomass for reuse. The effect of
desorption wash on structure of biosorbent has to be
established.

S. K. Mehta and J. P. Gaur 136

 IV. MODELING METAL SORPTION
A. Kinetics
The term “sorption kinetics” could be defined as
the velocity of adjustment of equilibrium between
metal concentration on biomass surface and metal
concentration in the surrounding fluid phase. Kinetic
model equations describe the quick kinetics of mass
transfer processes between fluid phase and solid sor-
bent phase as well as in both the phases. Differ-
ent kinetic models have been used to fit experimen-
tal data of heavy metal sorption (Ho, Wase, and
Forster, 1996; Mehta, Tripathi, and Gaur, 2002). To
analyze the rate of sorption of metal ions onto the
biomass, two simple kinetic models, namely pseudo-
first-order and pseudo-second-order models, are most
often used. The pseudo-first-order model considers
that the rate of occupation of binding site is propor-
tional to the number of unoccupied sites onto the
biosorbent. The pseudo-first-order rate expression is de-
scribed by the following equation (Ho and McKay,
1998):
dq
= k1,ads (qe − q) (2)
dt
where qe and q are the amount of metal ions adsorbed
on biosorbent at equilibrium and at any time t, re-
spectively (mg/g), and k1,ads (min−1 ) is the rate con-
stant of the first-order kinetics. Integration of the above
equation between the limits, t = 0 to t = t and q = 0 to
q = qe, gives Eq. 3:
k1,ads
log(qe − q) = log qe − t (3)
2.303
Linear plots of log (qe −q) versus t indicate the fitness of
sorption kinetics to the pseudo-first-order rate model.
The pseudo-second-order kinetics is applicable where
the rate of occupation of adsorption sites is propor-
tional to the square of the number of unoccupied
sites on the biosorbent. The second-order-rate kinetics
can be represented by the following equation (Ho and
McKay, 1998):
dq
= k2,ads (qe − q)2 (4)
dt
where k2,ads (g mg−1 min−1 ) is the rate constant of the
second order kinetics. Integration of the above Eq. 3
between the limits t = 0 to t = t and q = 0 to q = qe
137
gives:
1 1
= + k2,ads t (5)
qe − q qe
Straight line obtained after ploting t/q versus t shows
the degree of fitness of metal sorption data to the
pseudo-second-order rate kinetic model. It has been
shown that kinetics of metal sorption by algae fol-
lows the pseudo-second-order rate equation better than
the pseudo-first-order equation (Mehta, Tripathi, and
Gaur, 2002; Abu-Al-Rub et al., 2004; Cruz et al., 2004).
The kinetic rate constant (k2,ads ) of metal biosorp-
tion changes with different factors such as pH, initial
metal ion concentration, algal dose and temperature
(Cordero et al., 2004). Cordero et al. (2004) studied
the effect of various factors on kinetics of Cd sorp-
tion by Fucus spiralis, and found higher value of kinetic
constant at lower initial Cd concentrations and lower
pH of solution. However, kinetic constant increases
with increasing algal dose in solution (Cordero et al.,
2004).
In addition to simple kinetic equations, many other
mathematical models have been proposed incorporat-
ing various parameters. These models have been used
mainly for metal adsorption by chemical sorbents and
also by inactivated (dead) biomass, but have limited
utility for live biomass due to energy-driven intracel-
lular accumulation. These kinetic models are based on
transfer of mass in liquid phase, between liquid phase
and sorbent particle (film diffusion), and inside the sor-
bent particle (pore and surface diffusion). Correspond-
ing to different kinds of resistance taking place dur-
ing mass transfer process, the surface diffusion model,
the pore diffusion model, and combinations of both
have been used for describing the kinetics of adsorp-
tion on solid surfaces (Hu and Reeves, 1997, and refer-
ences cited therein). Nonequilibrium lumped-interior
model is an example where mass transfer coefficient de-
scribes both the film and the pore diffusion resistance.
Assuming that biosorption is limited by external surface
(of biosorbent), the rate of metal sorption can be de-
scribed by the following mass-transfer equation (Aksu,
1998):
dC
− = kj a(C − Ceq ) (6)
dt
where C is the residual metal ion concentration in the
solution at time t, a is the specific surface area of the
alga, kj is mass transfer coefficient, and Ceq is residual
metal concentration in solution at equilibrium.
Use of Algae for Removing Heavy Metal Ions From Wastewater
 Since the rate of metal sorption is limited by film
diffusion (mass transfer from the bulk phase to the ex-
ternal surface of the biosorbent), the following equation
could be used to estimate the rate of sorption by a sor-
bent particle (Bunke, Götz, and Buchholz, 1999):
δ q̄i
= kL.i as (Ci |r=RP − Ci (q̄i )) (7)
δt
where q is solid phase concentration (mol l−1 ), t is time,
kL is film diffusion coefficient (cm s−1 ), as is sorbent
particle surface area per sorbent particle volume (cm2
cm−3 ), C is metal concentration in solution (mole l−1 ),
r is radial distance (cm) and RP is particle radius (cm).
The film diffusion coefficient kL can be experimentally
determined.
Pore diffusion is another important factor that influ-
ences the kinetics of metal sorption by relatively larger
sorbent particles where intra-particular diffusion plays
an important role. Pore diffusion is the transport mecha-
nism where mass transfer in fluid filled pores of sorbent
particles takes place by diffusion of dissolved compo-
nent in the pore liquid inside the particles. Transport in
pores can also take place on inside surfaces. This trans-
port mechanism is termed as surface diffusion, and is
expressed by the equation (Bunke, Götz, and Buchholz,
1999):
 
δ q̄i δ 2 q̄i 2 δ q̄i
= DS· i + (8)
δt δr2 r δr
where DS· i is surface diffusion coefficient (cm2 s−1 ), r is
radial distance (cm); other symbols are as described in
Eq. 7.
The rate of metal sorption is influenced by the sorp-
tion equilibrium into active centers of the sorbent. This
influence can be evaluated by considering two rate con-
stants, one for sorption and the other for desorption,
and by assuming saturation in sorption. Metal sorption
and desorption rate constants can be determined using
equations given in Bunke, Götz, and Buchholz (1999).
The rate of metal adsorption onto the biosorbent is
also affected by metal concentration in the solution.
Chu and Hashim (2004) observed that film diffusion
is the rate-limiting step at low initial Cu concentration,
whereas intrinsic adsorption kinetics and film diffusion
are likely to control the overall rate of adsorption onto
Chlorella vulgaris at high initial concentration of Cu.
The above kinetic models cannot be applied for
metal sorption by algae immobilized or entrapped in
S. K. Mehta and J. P. Gaur
some matrix. Immobilization creates resistance in trans-
port of metal ions from bulk solution to the sorbent.
Entrapment of biosorbent particles in an immobiliza-
tion matrix may result in a decreased rate of metal sorp-
tion. Indeed, it has been earlier shown that the rate
of metal sorption is lower in immobilized cells than
the free cells. Metal sorption by immobilized biomass
could be represented by an equation that considers both
pore diffusion and surface diffusion mechanisms for
intra-particle metal transport. Such a model describ-
ing metal sorption by immobilized biomass bead has
been given in Hu and Reeves (1997). It should be kept
in view that the above models are not relevant where
ion exchange is the predominant mechanism of metal
sorption.
Kinetic coefficients derived from different models
are tools of engineering significance and are needed
for optimizing the metal removal process. They are re-
quired to test the appropriateness of the existing kinetic
models in describing metal sorption by algal biomass.
Further, it is imperative to develop some newer kinetic
models that can adequately characterize metal sorption
by algal biosorbents.
B. Sorption Models for Batch
Reactors at Equilibrium
Several mathematical models have been used for de-
scribing equilibrium metal sorption by algae in batch
system. Assuming adsorption and ion exchange as the
main mechanisms of metal sorption by algal biosor-
bent, researchers have proposed adsorption and ion ex-
change models, respectively (Özer et al., 1994; Volesky
and Holan, 1995).
1. The Langmuir Model
The Langmuir isotherm for single metal sorption by
algae can be expressed as (Langmuir, 1918):
 
bCeq
q = qmax (9)
1 + bCeq
where qmax is the maximum amount of metal ion ad-
sorbed at Ceq (equilibrium concentration of metal), and
b is a measure of the biosorption affinity or efficiency
of the biomass. The parameter qmax represents the maxi-
mum sorption capacity of the biosorbent when surface
is fully occupied by metal ions. It also represents to-
tal number of binding sites (per unit weight) involved
in metal sorption by a biosorbent (Crist et al., 1988).
138
The Langmuir model assumes (Langmuir, 1918): (i) a
monolayer adsorption of metals on binding sites; (ii)
all adsorption species (metal ions) interact only with a
site and not with each other; and (iii) adsorption en-
ergy of all the sites is identical and independent of
the presence of adsorbed species on neighboring sites.
The Langmuir model shows a good fit where passive
sorption of metals on biosorbents prevail (Aksu, 1998).
An important point to be noted here is that most of
the above assumptions are not fulfilled in biosorption.
Opposite to Langmuir assumptions, biological surfaces
have more than one type of binding sites contribut-
ing to biosorption process, each of which has a differ-
ent affinity for sorbing heavy metal ions. Furthermore,
Langmuir model assumes metal binding to free binding
sites rather than ion exchange. Therefore, for biosorp-
tion where ion-exchange is a major mechanism, Lang-
muir model is less appropriate (Crist et al., 1994).
2. The Freundlich Model
The Freundlich model of metal sorption by algae is
expressed by the equation (Freundlich, 1907):
1 complete monolayer on the surface. The BET model
q = kF Ceq
n
(10)
where kF is the Freundlich constant related to sorption
capacity of biomass and (1/n) is indicator of sorption
intensity. The Freundlich equation is empirical and de-
scribes multi-layered adsorption of metal ions on sor-
bent surface. The Freundlich model describes metal
sorption as a function of metal concentration in so-
lution at equilibrium, without reference to pH or other
ions in the same aqueous system. The Freundlich model
provides a more realistic description of metal adsorp-
tion by organic matter because it accounts for sorption
to heterogeneous surfaces or surfaces supporting sites
of varied affinity. In contrast to Langmuir model, the
Freundlich model does not assume saturation of metal
sorption. The Freundlich model assumes that stronger
binding sites onto biosorbent surface are occupied first
and that the binding strength decreases with increasing
degree of site occupation by metal ions.
For fitting the model to experimental data, the
Freundlich model generally gives a better fit for higher
equilibrium concentration of metal in solution. How-
ever, the application of these two models in interpreting
metal sorption by algae has met with some problems.
For example, Hashim et al. (1997) report that a unique
isotherm equation cannot be obtained if different initial
metal concentrations and algal doses are used for the
139
same set of experiments. Despite the fact that both the
models are used to fit a majority of experimental data
of biosorption, the Langmuir and Freundlich models
only reflect the influence of metal concentration on
the sorption of one particular metal, without any con-
sideration to important parameters such as pH, biomass
concentration, ionic strength or presence of other metal
ions.
In addition to the Langmuir and Freundlich models,
the Brunauer-Emmett-Teller (BET) equilibrium model
has been used for describing biosorption of metal ions
(Prasher et al., 2004). The BET model is expressed by
the following equation (Brunauer, Emmet and Teller,
1938):
BQCf
(11)
(Cs − Cf )[1 + (B − 1)(Cf /Cs )]
where Cs is the saturation constant of the solute, B is
a constant relating to the energy of interaction with
the surface, and Q is the number of moles of solute
adsorbed per unit weight of adsorbent in forming a
represents a multi-layered biosorption of heavy metals,
where the Langmuir equation could be applied for in-
dividual layer. The BET model further assumes that a
given layer need not complete formation prior to initi-
ation of subsequent layers.
Recently, Gin, Tang, and Aziz (2002) have derived an
equilibrium model through thermodynamic approach
for describing the relationship between important pa-
rameters for heavy metal sorption. In this model, both
the removal efficiency of heavy metal and metal sorp-
tion per unit algal biomass are considered to be simple
functions of the ratio of algal biomass concentration to
the initial metal concentration for selected conditions,
i.e., at constant pH and temperature. Model equation
is presented as:
 
Ce M
= α exp β (12)
Co Co
where Co and Ce are the initial and equilibrium con-
centrations of heavy metal in the solution, and M is
the concentration of algal biomass.α and β are coeffi-
cients. From the above, equation Eq. (13) can be de-
rived, which represents the relationship between metal
content per unit algal biomass (x/m) and the ratio of
metal concentration at equilibrium (Ce ) to initial metal
Use of Algae for Removing Heavy Metal Ions From Wastewater
concentration (Co ):
x 1−Ce /Co
=β (13)
m ln(Ce /αCo )
The above equation clearly shows an important differ-
ence from the widely used Freundlich and Langmuir
sorption isotherms. This model considers equilibrium
and initial metal concentrations rather than a func-
tion of equilibrium metal concentration alone. There-
fore, the above model can be used under both equilib-
rium and non-equilibrium conditions. In this model,
the value of β reflects the metal sorption ability of the
biomass. The higher the absolute value of β, higher is
the sorption ability of the biosorbent. The coefficient
α is equal to 1 under ideal conditions, and under actual
experimental conditions it may take on values either
larger or smaller than 1. The above model can be de-
rived to represent the effect of pH on sorption of metal
as well (Gin, Tang, and Aziz, 2002). Although the above
model is not tested by other researchers, Gin, Tang, and
Aziz (2002) found the above model to fit well for ex-
perimental results of Cd, Cu, Pb and Zn adsorption by
two algal species, Oocystis sp. and Chlorococcum sp. (live
as well as dead). The above authors pointed out that
this model is not appropriate at very low and high pH
levels. As this model was derived through a thermody-
namic approach, it does not provide any information
on actual mechanism of metal sorption by algae.
3. Modeling Metal Sorption From
Metal Mixture
Because wastewaters contain more than one metal
ion simultaneously, both Langmuir and Freundlich
models cannot be used to describe the metal sorp-
tion behavior of biosorbents. When a solution contains
several components, for instance metal ions, there oc-
curs interference and competition for adsorption sites.
Hence, metal sorption from a multicomponent system
needs more complex mathematical models for describ-
ing the equilibrium sorption data. The following are
some complex models used for describing metal sorp-
tion from multi-metal systems:
a. Three-Parameter Extended Langmuir
Model
Metal sorption form the binary metal system could
be described by the three-parameter extended Langmuir
S. K. Mehta and J. P. Gaur
equation given below (Al-Asheh and Duvnjak, 1998):
qmax1 b1 Cf1
q1 = (14)
1 + b1 Cf1 + b2 Cf2
where q1 is the equilibrium sorption (mmol g−1 ) of the
first metal, qmax1 is the maximum sorption of the first
metal at equilibrium concentration of metal in solu-
tion (Cf1 and Cf2 , mM); b1 and b2 are the constants
(l mmol−1 ) for the first and the second metal, respec-
tively; subscripts 1 and 2 represent the metal of primary
interest and the second metal, respectively. Sorption of
the second metal q2 can also be represented in the same
way using the above equation, and qmax2 can be deter-
mined.
b. Multicomponent Extended Langmuir
Model
Metal sorption from multi-metal mixture could be
evaluated by using multicomponent extended Lang-
muir model with four parameters as given below
(Chong and Volesky, 1996):
q(Mi )
(qmax /Ki )Cf [Mi ]
=
1 + (1/K1 )Cf [Mi ] + (1/K2 )Cf [M2 ] + (1/K3 )Cf [M3 ]
(15)
where i (1 to 3) represent metal in question. Cf [M1 ],
Cf [M2 ] and Cf [M3 ] are residual concentration of metal
1, 2 and 3, respectively. The four parameters (qmax , K1 ,
K2 and K3 ) of the above model could be evaluated using
an appropriate software package.
Although the above model is empirical, it is very
often used to describe the equilibrium isotherm data of
multicomponent systems.
c. Langmuir-Freundlich Model
qmax1 b1 Cnf11
q1 = (16)
1 + b1 Cnf11 + b2 Cnf22
The above model is a combination of the conventional
Langmuir and Freundlich models (Al-Asheh and Duvn-
jak, 1998). Five parameters of the above model (qmax , b1 ,
b2 , n1 and n2 ) for sorption of metal ions from a multi-
component system could be obtained by fitting the data
using a suitable software. In the same way, sorption of
the second metal (q2 ) could also be represented.
140
4. Ion Exchange Models
Several researchers have independently concluded
that the major mechanism of heavy metal sorption by
algae is ion exchange (Crist et al., 1990; Kratochvil,
Volesky, and Demopoulos, 1997). Therefore, some
ion exchange models have been proposed (Kratochvil,
Volesky, and Demopoulos, 1997) to fit and interpret
the data obtained from equilibrium as well as dynamic
biosorption experiments. One of these models, called
the ion-exchange isotherm model, represents ion ex-
change reaction between metal species, M1 and M2
(metal ions or cations bound on biosorbent (X)). Ion
exchange reaction is given as:
X − mM2+n + nM1+m ←→ X − nM1+m + mM2+n
(17)
For the above reaction equilibrium constant is rep-
resented as:
qn M1CmfM2
KM1M2 =
qm C n
M2 fM2
Assuming the above ion exchange reaction, the ion
exchange isotherm equation can be derived (Figueira
et al., 2000):
Q + St (KSM [M]0.5 )/(1 + KSH [H] + (KSM [M])0.5 )
qM1 = (18)
1 + CfM2 /KM1M2 CfM1
Q
qM2 = (19)
1 + CfM2 /KM1M2 CfM2
where M1 and M2 are heavy metal ions to be sorbed
and already bound on the biosorbent, respectively. m
and n are the valences of M1 and M2, respectively; Q is
the total number of binding sites in algal biomass; CfM1
and CfM2 are the equilibrium concentrations of metal
M1 and M2 in solution, respectively. qM1 and qM2 are
the content of M1 and M2 in the biosorbent. In this
model M2 can be replaced by proton [H] if biomass is
protonated.
The ion-exchange model is certainly a better repre-
sentation of metal sorption, and it reflects that algal
biomass is protonated or contains light metal ions (K+ ,
Na+ , Mg2+ ) that are released upon binding of heavy
metal cations. However, ion-exchange isotherm did not
account for influence of important variables such as
change in initial heavy metal concentration and al-
gal biomass concentration in solution, pH and ionic
strength. The above ion-exchange isotherm could not
141
be applied when it is considered that more than one
ionic species are bound to the algal surface (Gin, Tang,
and Aziz, 2002).
There are many equilibrium models, which incor-
porate metal ion concentration, pH, ionic strength
and biomass swelling (Schiewer, 1999; Schiewer and
Wong, 2000). One of them is the two-site model
for predicting ion exchange and pH effect (Schiewer,
1999). This model is based on the fact that in ion-
exchange process 2H+ ions are released from pre-
viously protonated biomass for each divalent metal
ion bound. Two-site ion exchange model assumes
two types of binding sites, carboxyl (C) and sulphate
(S) groups to be present in the biomass. Consider-
ing metal and proton binding on these two sites,
Schiewer and Volesky (1995) derived an isotherm equa-
tion that is able to predict the binding of protons and
metal ions as a function of metal concentration and
pH.
qH = Ct (KCH [H])/(1 + KCH [H] + (KCM [M])0.5 )
+ St (KSH [H])/(1 + KSH [H] + (KSM [M])0.5 )
(20)
0.5 0.5
qM = Ct (KCM [M] )/(1 + KCH [H] + (KCM [M]) )
(21)
where Ct and St are the total number of carboxylic and
sulfate sites, respectively. Constant KCH and KSH are
equal to 104.8 (pKa for carboxyl group, 4.8) and 102.5
(pKa for sulfate group, 2.5) (Schiewer, 1999). KCM and
KSM represent metal binding constant for binding of
metal (M) on carboxyl and sulfate groups, respectively.
qH and qM represent proton and metal uptake, respec-
tively, by biosorbent. Values of Ct and St need to be de-
termined for different algal biosorbent. Constants KCH
and KSH can be used for binding of any metal on a
biosorbent. Parameter KCM and KSM have to be deter-
mined for binding of individual metal. Although appli-
cability of this model has not been tested other than
by its developer, Schiewer (1999) pointed out that be-
ing pH sensitive, two-site model could successfully pre-
dict the binding of both protons and metal at different
pH values and metal concentrations. Because two-site
model considers the effect of pH and proton binding,
it has a better predictive power than Langmuir model.
Use of Algae for Removing Heavy Metal Ions From Wastewater
 V. CONTINUOUS SYSTEMS FOR
METAL REMOVAL
A large number of metal biosorption studies have
been performed in batch system. In a batch reactor,
algal biomass is kept in a metal containing solution
for a time sufficient for saturation of metal binding
by the biomass. Mixing of biomass and metal solution
is done either by air or a magnetic stirrer. Biomass is
separated from metal solution by centrifugation or fil-
tration. Metal-loaded biomass can be regenerated for
repeated use in multiple sorption-desorption cycles or
disposed off in an environmentally acceptable manner
(Pradhan and Rai, 2001). However, biomass separation
by centrifugation or filtration will not be practical in
industrial processes where large volumes of effluent are
to be treated. Basic factors like properties of biosorbent
surface and physicochemical characteristics of the envi-
ronment, as described in the previous section, control
the removal of metal ions by algal biosorbent in a batch
reactor. There are many other variables that could be
considered in selection and designing of a batch reactor
for the removal of metals from solution (Aksu, 1998).
Batch reactor is suitable where the living biomass is used
for metal removal.
Separation of biomass from the liquid phase is one
of the major problems associated with batch reactors,
and hence environmental engineers prefer a continu-
ous system of metal removal. Algal biomass, preferably
immobilized, can be used just like ion exchange resins
in continuous systems, such as continuous stirred tank
reactors, fluidized bed, moving bed and packed bed
columns (Fig. 3). For continuous operation, the most
convenient configuration is that of a packed column. A
packed bed column system could be constructed by a
piece of glass column or PVC pipe stood on its end and
filled with a biosorbent either in free or immobilized
form. In up-flow column the fluid to be treated flows
upward, and in the down-flow column the solution
flows down into the column. When a metal containing
solution passes through a packed bed column, most of
the metal ions get adsorbed on biosorbent. Therefore
metal concentration in the effluent (outflow) remains
either very low or is not at all detectable in some cases.
As biosorption continues, the metal concentration in
effluent rises, slowly at first, and then abruptly. When
this abrupt rise or “breakthrough” occurs, the flow is
stopped. The breakthrough point is an important pa-
rameter for evaluating metal sorption performance of
S. K. Mehta and J. P. Gaur
a biosorbent in a continuous operation system. For
the evaluation of column performance, breakthrough
curves are plotted. A breakthrough curve shows rela-
tion between metal concentration in effluent (outgoing
solution) and breakthrough time or volume. The posi-
tion of the breakthrough point on volume or time axis
depends on the capacity of the column with respect to
feed concentration and flow rate.
Figure 4 represents different kinds of breakthrough
curves showing the ratio (Ci /Ce ) of metal concentration
in influent (Ci ) and effluent (outgoing) solution (Ce )
on y-axis and the volume of solution passed through
the column on x-axis. Arrows show the position of
breakthrough points. The performance of a biosorbent
packed column is proportional to the distance of break-
through point from the origin on x-axis. Curve (1) shows
that a relatively larger volume of wastewater, compared
to other curves, could be treated before the onset of
breakthrough point. For curve (5) the breakthrough
point is very close to the origin, thereby indicating that
a relatively smaller volume could be treated by this sys-
tem than the others. After breakthrough point, all the
curves show a rise in Ci /Ce . The steepness of the curve
also indicates the efficiency of a column. A packed bed
column with breakthrough curve showing a slow rise in
Ci /Ce is preferred over the one having a breakthrough
curve with a sharp rise in Ci /Ce . For curve (3) and (2) the
breakthrough points are the same but the rise in Ci /Ce
is sharper for curve (3) when compared with curve (2). It
shows that column (3) becomes exhausted earlier than
column (2), and therefore the latter column is more
efficient than the former. Solid vertical lines in Fig. 4
show saturation points for different columns. At satu-
ration point, there is no metal sorption and the Ci /Ce
becomes equal to unity. Sometimes Ci /Ce in effluent
solution exceeds 1.0 (curve 4), thus indicating that after
saturation there is desorption of metal ions from the
metal-loaded biomass filled in the column. For all prac-
tical purposes, the column should not be used after the
breakthrough point is reached.
A large number of trials have been made to remove
metals from synthetic metal solutions as well as indus-
trial effluents in continuous systems using algae (see
Aksu, 1998). In column experiments, 99% removal of
metals from solution can be obtained before the break-
through point is reached. Packed-bed reactors should be
optimized for operational conditions like flow rate, bed
height, matrix type, biomass loading and metal concen-
tration prior to large-scale application. In these reactors,
142
FIGURE 3 Basic designs of reactors for continuous removal of heavy metals from wastewaters.

however, the level of metal accumulated in algae re- the film and pore diffusion resistance to mass trans-
mains lower than batch systems (Wilkinson, Goulding, fer (Aksu, 1998). In continuous fixed bed biosorption,
and Robinson, 1989, 1990). the metal concentration in the fluid and solid phase
Operational parameters, such as pH, influent metal changes with time as well as with the position in the
concentration, size of biosorbent particles and column bed. For a continuous packed bed column the mass
length, are important in in controlling metal removal balance for the metal ions could be described by the
in packed bed columns (Yu and Kaewsarn, 1999; Vija- following equation (Aksu, 1998):
yaraghavan et al., 2005). In order to scale-up parameters
for the design of the reactor and to determine oper- ∂C ∂C ∂ 2C ∂q
ational conditions, some mathematical models have ε = −ν + E 2 − (1−ε) (22)
∂t ∂z ∂r ∂t
been developed for metal removal in continuous sys-
tems. One such model is “the distributed parameter where C is residual metal concentration in the fluid
model,” which has been developed taking into account system, ε is void fraction in the bed, ν is superficial

143 Use of Algae for Removing Heavy Metal Ions From Wastewater

FIGURE 4 Diagrammatic presentation of different kinds of
breakthrough curves (1–5) for the continuous removal of heavy
metals from wastewaters using biomass packed column. Ci /Ce is
the ratio of metal concentration in influent (Ci ) and effluent (Ce )
solution. The volume of the solution passed through the column
is shown on x-axis. Arrows show breakthrough points and vertical
lines indicate saturation points.
velocity, E is the axial dispersion coefficient, t is the
time, and r is axial coordinate.
VI. METAL REMOVAL
BY PERIPHYTON
Periphyton is an assemblage of algae, bacteria and
other microorganisms that grows attached to sub-
merged surfaces in streams and other shallow waters.
Periphyton often contains a lot of exopolymers that
help in attachment of organisms to the surface. Peri-
phyton is therefore a kind of self-immobilized mixed
culture. Algal periphyton display stable operating per-
formance, good mechanical stability and biomass/fluid
separation, and excellent ability to take up nutrients
and pollutants from water. In fact, a novel technology
called “algal turf scrubbing” has been developed that
utilizes the periphytic organisms for the treatment of
agricultural run off and eutrophic waters (see Craggs
et al., 1996). Algal turf scrubber (ATS)TM consists of a
mixed periphytic assemblage that colonizes an inclined
floway over which wastewater flows in pulses. However,
this technology has not been widely used for the re-
moval of metals from wastewater, although there are re-
ports showing tremendous ability of periphytic algae in
sorbing metal ions from the solution (Vymazal, 1984;
S. K. Mehta and J. P. Gaur
Newman and McIntosh, 1989; Gray and Hill, 1995;
Gray, Hill, and Stewart, 2001; Schumacker, Blume, and
Sakoulov, 2003; Bender and Phillips, 2004).
Algal periphyton may be a few cell layers to a few mm
thick (film), or may be <1 mm to a few cm thick (mat).
However, mats and films share many characteristics,
and both represent communities with complex strate-
gies for microorganisms at surfaces where steep gradi-
ents of physicochemical variables are found. Algal mat
and biofilm have been shown to sequester heavy met-
als, metalloids, radionuclides and oxyanions (Bender
and Phillips, 2004, and references cited therein). These
communities use a variety of mechanisms for sequestra-
tion of metal contaminants (Bender and Phillips, 1994).
Phillips and Bender (1998) have shown that a micro-
bial mat accumulated Mn and Zn up to 12,050 and
30,300 mg kg−1 , respectively. In another study, micro-
bial mat immobilized on silica particles removed 88%
U6+ (initial concentration, 2.4 mg l−1 ) from drainage
water within 15 min (Bender et al., 2000). Liehr, Chen,
and Lin (1994) developed in the laboratory a biofilm of
Cladophora glomerata using 0.4 µm polycarbonate mem-
brane filters and placed it in batch reactor to determine
metal sorption by the biofilm. They have shown that
the algal film could take up about 2200 µg Cu g−1 its
dry weight. Liehr, Chen and Lin (1994) suggested that
increased metal sorption is due to higher pH inside
the biofilm. Good evidence exists to support the con-
cept of pH variation within photosynthetic biofilms.
The pH inside the algal biofilms and mats can be
measured with a microelectrode (Revsbech et al., 1983;
Kuenen, Jorgensen, and Revsbech, 1986; Revsbech and
Jorgensen, 1986). The pH inside algal biofilms can be
several units higher than the pH of the surrounding
liquid (Kuenen, Jorgensen, and Revsbech, 1986; Liehr,
Chen, and Lin, 1994). The nature of the algal biofilm
is such that organisms grow very closely attached to
each other, thus providing resistance to mass transfer
of photosynthetically generated CO2 out of the film.
This leads to higher pH inside the biofilm than the
surrounding milieu. The pH greatly affects metal speci-
ation, binding sites on algal surfaces, and precipitation
of metal compounds inside algal biofilm. The increased
pH may result in increased precipitation of metals in-
side the biofilms and mats, although some other mech-
anisms may also be important (Liehr, Chen, and Lin,
1994). The increased pH inside the biofilm may also in-
crease its metal sorbing capability but it needs to be val-
idated experimentally. One attractive feature of metal
144
removal by algal biofilm is that it would be very easy
to recover the metal from the biofilm by introducing
CO2 inside the reactors. This could be used for metal
recovery and reuse or for simply concentrating metals
in a liquid stream.
Metal sorption by the periphyton greatly depends
on developmental time, taxonomic composition, abi-
otic components (oxides of Fe and Mn), and light in-
tensity (Gray and Hill, 1995; Gray, Hill, and Stewart,
2001). Therefore, a detailed study of the above factors
could help in selecting suitable periphyton biomass for
the removal of heavy metals from industrial effluents.
Inorganic constituents of periphyton could offer the ad-
vantage of chemical sorption in addition to biosorption
by periphyton (Newman, McIntosh, and Greenhut,
1983; Newman, Alberts, and Greenhut, 1985). Periphy-
ton often contains substantial levels of hydrous iron
(Fe) and manganese (Mn) oxides, clay and other inor-
ganic particles. These inorganic compounds may also
strongly sorb metals (Jenne, 1968; Kinniburgh and Jack-
son, 1981).
VII. NON-BIOSORPTIVE REMOVAL
OF HEAVY METALS BY ALGAE
Van Hille et al. (1999) used alkalinity generated by
the cyanobacterium Spirulina sp. for precipitating heavy
metals from effluents. The idea is based on two well-
established facts: (i) algae and cyanobacteria increase
the alkalinity of the surrounding medium as a by-
product of their inorganic concentrating mechanism
(Shiraiwa, Goyal, and Tolbert, 1993), and (ii) the sim-
plest and cheapest method for removing heavy metals is
to increase the pH of the medium so as to convert met-
als into insoluble forms, like hydroxides. The method
developed by Van Hille et al. (1999) separates the algal
system from metal-containing stream so as to obviate
the problem of metal toxicity to algae. Spirulina was al-
lowed to grow in a tank, and this led to increase in pH to
over 9. Spirulina filaments were separated and overflow
from the algal tank was mixed with metal-laden water so
as to precipitate heavy metals. The treatment process re-
moved over 99% iron (98.9 mg l−1 ), and 80–95% of Zn
(7.16 mg l−1 ) and Pb (2.35 mg l−1 ) during a 14-day time
period, during which pH of the effluent increased from
1.8 to over 7. Although the system is efficient, it is non-
selective in so far as precipitation of metals is concerned.
Some algal species have shown the capability of non-
sorptive removal of mercury from aqueous medium by
145
volatilization (De Filippis and Pallaghy, 1976; Wilkin-
son, Goulding, and Robinson, 1990; Devars et al., 2000).
De Filippis (1978) reported that volatilization of mer-
cury by Chlorella sp. required an enzyme system, sul-
phydryl groups and NADPH/NADP as the energy
source. Mercury volatilization by algae depends on cell
density, mercury concentration in the medium, and the
process is light dependent (Devars et al., 2000). How-
ever, the process is not environmentally sound because
the release of mercury into the atmosphere may lead to
serious ecological repercussions.
VIII. ELECTROLYSIS: THE LAST STEP
OF THE METAL REMOVAL PROCESS
A logical approach to the treatment of wastewater
could be to combine removal process with metal re-
covery. This practice is particularly necessary when a
wastewater contains precious metals like Au and Ag, etc.
Metals can be recovered by electrolysis of eluate result-
ing from desorption process. For electrolysis of metal
containing eluate, cathode cells may be used. Low levels
(>10 mg l−1 ) of metals in eluate could create some diffi-
culty in electrolysis that may be overcome with rotating
cathode cells (Walsh, 1992). Rotating cathode cells have
been successfully used in electrolysis of eluates contain-
ing Cd as low as 10 mg l−1 (Butter et al., 1998). After
separation of metal, desorbing solution could be used
for the next cycle of desorption. However, more studies
are needed in this direction.
IX. EPILOGUE
The ability of algae to sorb high concentrations of
heavy metals makes them a suitable candidate for re-
moving these ions from wastewaters. This might be-
come possible if an efficient and commercially viable
algal technology is developed for metal removal from
wastewaters. Development of such a technology would
require filling in a number of gaps in knowledge con-
cerning metal sorption by algae. There is a need to
develop a thorough understanding of the mechanism
of metal sorption. Metal sorption by algae has been
generally described using Langmuir and the Freundlich
models. However, it is necessary to develop a model
that takes into account the parameters, such as pH
and biomass concentration, size of sorbent particles,
etc., in order to predict the metal sorption capacity
of the biomass. Despite the fact that industrial efflu-
ents contain several metal ions simultaneously, little
Use of Algae for Removing Heavy Metal Ions From Wastewater
attention has been given to sorption of metals from
multi-metal solutions. Many abundantly growing fresh-
water and marine algae have not been evaluated for
their metal binding capacity. Therefore, screening of al-
gae should be done in order to select species having a
high affinity for a particular metal or species able to ef-
ficiently bind several metal ions concomitantly. Once
suitable metal sorbing species are identified, the tech-
nology for metal removal should be perfected. In its
native form, algal biomass cannot be used for remov-
ing metal ions in a continuous system as biomass sep-
aration would be virtually impossible, and moreover it
may clog the column. Therefore, biomass has to be im-
mobilized or granulated so that it can be filled in the
column for passing wastewater through it. Immobiliza-
tion of biomass with expensive chemicals (e.g., alginate)
is not feasible, and therefore naturally immobilized al-
gae hold tremendous potential for metal removal from
wastewater. In this context, algal biofilms and mats,
seem particularly attractive for removing metals from
solution.
It may be possible to produce genetically engineered
strains of algae with increased synthesis of metalloth-
ioneins or phytochelatins eventually leading to en-
hanced metal binding capacity (Gardea-Torresdey et al.,
1998). Another probability could be genetic manipula-
tion of algae for increased synthesis of cell wall polysac-
charides and other metal-binding sites on the cell sur-
face for increasing their metal removal potential. Algal
technology for metal removal may eventually turn out
to be more efficient than the conventional physico-
chemical methods. However, comparative economics
of algal and physicochemical methods of metal removal
from wastewater have not been critically worked out.
Indeed, a lot of efforts have been made and some tech-
nologies have even been patented, but there is still a
long way to go in fully exploiting the metal sorption
potential of algae.
ACKNOWLEDGEMENTS
We are grateful to the Head, Department of Botany,
and the Coordinator, Centre of Advanced Study in
Botany, Banaras Hindu University, for facilities, and
Dr. B.A. Whitton, University of Durham (U.K.), for
useful suggestions. SKM thanks the Council of Scien-
tific and Industrial Research, New Delhi, for financial
support.
S. K. Mehta and J. P. Gaur
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