Chemical Engineering Journal 168 (2011) 737–745

Contents lists available at ScienceDirect

Chemical Engineering Journal

journal homepage: www.elsevier.com/locate/cej

Study on the adsorption of Ca2+ , Cd2+ and Pb2+ by magnetic Fe3 O4 yeast treated
with EDTA dianhydride
Meng Xu a,1 , Yunsong Zhang a,1 , Zhiming Zhang b , Yaou Shen b , Maojun Zhao a,∗ , Guangtang Pan b,∗
a
College of Life and Science, Sichuan Agricultural University, Yaan 625014, PR China
b
Maize Research Institute, Sichuan Agricultural University, Yaan 625014, PR China

a r t i c l e i n f o a b s t r a c t

Article history: Magnetic Fe3 O4 baker’s yeast biomass (FB) was prepared by combining baker’s yeast biomass and
Received 18 November 2010 nano-Fe3 O4 using glutaraldehyde as a cross-link agent, and was chemically treated with ethylene-
Received in revised form 17 January 2011 diaminetetraacetic dianhydride (EDTAD). The EDTAD-treated magnetic Fe3 O4 baker’s yeast biomass
Accepted 17 January 2011
(EFB) was investigated by Fourier transform infrared spectroscopy (FTIR), potentiometric titration, zeta
potential, and magnetic response analysis. The results revealed that the EFB possessed not only the super-
Keywords:
paramagnetic characteristic of nano-Fe3 O4 , but its surface also had plenty of carboxyl and amino groups
Baker’s yeast
introduced by the EDTA molecules. The adsorption properties of EFB for Pb2+ , Cd2+ , and Ca2+ ions were
Nano-Fe3 O4
Super-paramagnetic
also evaluated. The results showed that the uptakes of EFB for the three metal ions were higher than
EDTAD that of FB, and the adsorption capability of Pb2+ , Cd2+ , and Ca2+ ions increased with an increase in pH.
Chemically treated The adsorption process was followed by the pseudo-second-order kinetic model and Langmuir isotherm
Adsorption equation. The maximum adsorption capacities of 99.26 mg/g for Pb2+ at pH 5.5, 48.70 mg/g for Cd2+ at pH
6.0, and 33.46 mg/g for Ca2+ at pH 6.0 were observed at 30 ◦ C. The regeneration experiments showed that
the EFB could be successfully reused.
© 2011 Elsevier B.V. All rights reserved.

1. Introduction agricultural production [13,14]. Saccharomyces cerevisiae, which

Water pollution, especially heavy metal ion pollution, is cur-
rently one of the most serious environmental problems [1,2]. Heavy
metal ions such as lead, copper, cadmium, zinc, nickel, and oth-
ers are the most common pollutants found in industrial effluents
[3]. They are non-biodegradable, and thus some heavy metal ions,
even at low concentrations, are highly toxic to living organisms
[4]. Among them, lead and cadmium can directly damage the ner-
vous, reproductive, and skeletal systems and the kidney, and can
even cause cancer. Thus, finding an efficient method that can purify
waste water is necessary. Although there are traditional methods,
such as chemical precipitation, ion exchange, electrolysis, filtra-
tion, extraction, and evaporation, among others [5–8], these are
sometimes restricted by practical technique or economic factors
[9].
Recently, the utilization of biomass and agricultural waste mate-
rials for the removal of metal ions has been explored [10–12].
Attention has been diverted toward the usage of biomaterials,
which are byproducts or wastes from large-scale industrial and
∗ Corresponding authors. Tel.: +86 835 2885782; fax: +86 835 2862227.
E-mail address: numberlala123@163.com (M. Zhao).
1
These authors contributed equally to this work.
has been considered a cheap, available, and safe industrial
microorganism, is an economic and constant supply source of
microorganism for the removal of heavy metal ions [15,16]. Inves-
tigations conducted by many researchers have demonstrated that
baker’s yeast biomass is capable of accumulating heavy metals,
such as Cu2+ , Cd2+ , Pb2+ , Hg2+ , and others from waste water [7].
However, because the adsorption capability of heavy metal ions by
waste brewery biomass is lower, improving the removal efficiency
of the yeast biomass by chemical treatment methods becomes the
focal point in recent research [9,17].
Ethylenediaminetetraacetic acid (EDTA), a powerful complex-
ing agent, is widely commercially available. EDTA, which is a Lewis
acid, has six binding sites, four carboxyl and two amino groups,
providing six pairs of electrons. The resulting metal–ligand com-
plex, where EDTA forms a cage-like structure around the metal
ions, is very stable. EDTA dianhydride (EDTAD), a ramification of
EDTA containing two anhydride groups per molecule, can react
with hydroxyl and amino groups of other materials. The use of
EDTAD to treat baker’s yeast biomass for producing biosorbents
with high adsorption capacity of Pb2+ and Cu2+ ions has been
reported in the study [18]. However, the separation of EDTAD-
treated baker’s yeast biomass from disposed waste water, as well
as the regeneration of the adsorbents, is very difficult in practical
application.

1385-8947/$ – see front matter © 2011 Elsevier B.V. All rights reserved.
doi:10.1016/j.cej.2011.01.069
738 M. Xu et al. / Chemical Engineering Journal 168 (2011) 737–745
Nanomaterials are widely considered as the new functional
materials; they have been utilized in various fields such as
medicine, environment, and industry [19,20]. Nano-Fe3 O4 , which
is superparamagnetic, presents high recovery ability [21]. To facil-
itate the recovery of baker’s yeast, Fe3 O4 nano-particles are loaded
on the baker’s yeast biomass, enabling it as an efficient candidate
for combining adsorption properties with ease of phase separation
in the removal of heavy metal ions.
In this work, the magnetic Fe3 O4 baker’s yeast biomass (FB) was
prepared using glutaraldehyde as a cross-link agent and was chem-
ically treated with EDTAD. The resulting EDTAD-treated FB (EFB)
was investigated by magnetic response, Fourier transform infrared
(FTIR) spectroscopy, potentiometric titration, and zeta potential
analysis. The mechanisms of EFB were studied. EFB was utilized as
an adsorbent for the removal of Ca2+ , Cd2+ , and Pb2+ in the solution.
The effects of the experimental parameters such as time, pH, tem-
perature, and initial concentration of the adsorbate on adsorption
were also investigated. The mechanism of interaction between the
functional groups on EFB and metal ions as well as the regeneration
properties of EFB was also clarified.
2. Materials and methods
2.1. Materials
Baker’s yeast was purchased from Harbin Mali Ltd., China. The
samples were repeatedly washed with deionized water to remove
dirt and soluble impurities. They were dried at 80 ◦ C for 24 h. After-
wards, they were crushed and sieved to a particle size of under 100
meshes using a standard sieve.
The stock standard solution of Cd(NO3 )2 , Pb(NO3 )2 , and
Ca(NO3 )2 with 1000 ␮g/mL Pb2+ , Cd2+ , and Ca2+ was obtained from
the National Analysis Center for Iron and Steel (Beijing, China).
The working solutions were diluted from the above stock solu-
tion. Other chemicals were of analytical grade and were purchased
from ChengDu KeLong Corp., China. Ultrapure water with resistiv-
ity of 18.23 M/cm obtained from a pure water system (Ai Kuo,
KL-UP-11-20) was used throughout the experiment.
2.2. Synthesis of EFB
2.2.1. Preparation of FB
The synthetic nano-Fe3 O4 was prepared according to Shan Zhi’s
method [22]. NaOH at 4 mol/L (100 mL) was placed in a three-
neck flask under nitrogen flow at 150 rpm stirring at 65 ◦ C for
30 min; 56 mL of solution containing Fe2 SO4 (0.01 mol) and FeCl3
(0.018 mol) was quickly added. The resultant mixture was stirred
for 90 min and was washed with ultrapure water until the pH of the
supernatant became neutral with the help of an external magnetic
separator. The resultant dark precipitate was placed in a vacuum
drying oven overnight until further use.
After 1.0 g of synthetic nano-Fe3 O4 was dispersed in 200 mL of
ultrapure water by ultrasonication for 20 min, 5.0 g of dry baker’s
yeast biomass and 100 mL glutaraldehyde solutions (1.5 wt% in
water) were added to the suspension; the mixture was stirred at
room temperature for 24 h. After the reaction, FB was obtained by
an external magnet and was washed thrice before it was freeze-
dried in high vacuum for 24 h. Finally, FB was preserved in a
desiccator for further use.
2.2.2. Synthesis of EDTAD
EDTAD was prepared following the methodology described by
Capretta et al. [23]. About 18.0 g EDTA was suspended in 30 mL
anhydrous pyridine. Then, 24 mL of acetic anhydride was added to
the suspension. The mixture was stirred at 65 ◦ C for 24 h. EDTAD
obtained was filtered. It was then washed with acetic anhydride
and diethyl ether before it was dried under vacuum and stored in
a desiccator.
2.2.3. EDTAD treatment of FB
Approximately 2.0 g of EDTAD was added to 100 mL of N,N-
dimethylformamide (DMF) containing 5.0 g of FB in a three-neck
round bottom flask equipped with a condenser. The mixture was
stirred at 60 ◦ C for 4 h. After the reaction, EFB was isolated from
the mixture by an external magnet and then washed with DMF,
ultrapure water, and 10% NaHCO3 solution, respectively. EFB was
freeze-dried in high vacuum for 24 h and then preserved in a des-
iccator for further use.
2.3. Biosorption studies
Batch biosorption studies were conducted in 100 mL conical
flasks at different temperatures (10, 20, 30, 40, 50, and 60 ◦ C).
Then, 100.0 mL of metal ion (Cd2+ , Ca2+ , and Pb2+ ) solution at
an initial concentration of 40–300 mg/L was stirred at 150 rpm
in a rotary shaker (DH2-DA, China) with 0.1 g dry sample for a
required biosorption time (5–120 min). Varying pH values of the
solutions (2–6) were used. Afterwards, the external magnetic field
was applied to separate the samples from the solution. The super-
natants were analyzed to determine the concentration of the metal
ions by an atomic absorption spectrometer (Japan Shimadzu AA-
6300). The adsorption capacity was calculated using the following
relationships:
(Ci − Ct )V
qt = (1)
m
Ci − Ct
Q (%) = × 100% (2)
Ci
where qt is the amount of adsorption capacity at time t (mg/g), Q
is the metal ions uptake efficiency, Ci is the initial concentration
of metal ions (mg/L), Ct is the metal ions concentration at time t
(mg/L), V is the volume of the solution (L), and m is the mass of the
adsorbent (g).
2.4. Desorption studies
Desorption studies were carried out in 100 mL conical flasks.
The metal ion-loaded EFB was magnetically separated. The set-
tled EFB was eluted with different eluents. The suspension was
shaken at 150 rpm for 30 min to release the metal ions from the
EFB. The desorbed metal ions were analyzed, and the metal ions
recovery efficiency was calculated. The EFB regeneration efficiency
was calculated as follows:
Metal desorbed efficiency
Amount of metal ions desorbed
= × 100% (3)
Amount of metal ions adsorbed
2.5. Characterization of EFB and FB
The zeta potential of EFB and FB (with/without metal ions) was
measured using a zeta potential analyzer (Malvern–Zetasizer nano
ZS, UK) at pH 5. The types of binding groups present in the EFB and
FB were identified by FTIR spectroscopy (Shimadzu FTIR-8400S,
Japan) analysis in the region of 400–4000 cm−1 through the KBr
pressed-disc method. The active sites present on the surface of
EFB and FB were determined by the potentiometric titration on
an autotitrator (ZD-2, China) with a combined glass electrode. The
software ProtoFit Version 2.0 [24,25], a useful software for the
calculation of pKa values and functional groups on the surface of
biological material, was employed to fit the acid–base titration data
of the FB and EFB.
 M. Xu et al. / Chemical Engineering Journal 168 (2011) 737–745 739

Fig. 1. Preparation scheme of EFB and the adsorption mechanism of heavy metal ions on EFB.

3. Results and discussion of the FB, EFB, and the 0.1 mol/L NaCl electrolyte. Based on the
figure, FB and EFB provided a significant buffering capacity to the
3.1. Mechanism of preparation and adsorption of metal ions on solution across the pH range studied compared with the blank. This
EFB buffering capacity was due to the functional groups on the biomass
surface consuming the added base by donating protons. However,
Fig. 1 presents the procedure for the synthesis of EFB in the both FB and EFB suspensions showed different titration curves that
present work and the suggested mechanism for the Ca2+ , Cd2+ ,
or Pb2+ adsorption onto the EFB. The synthesis of EFB was per-
formed in three steps. First, FB was synthesized by combining
baker’s yeast and Fe3 O4 nanoparticles using the glutaraldehyde
cross-linked method. Second, the FB was reacted with EDTAD in
anhydrous DMF, and the EDTA molecules were introduced onto the
FB surface through the esterification reaction. Third, the EFB was
rinsed with a NaHCO3 solution; the additional exposed functional
groups, such as carboxyl and amino groups, on EFB could improve
the surface activity and increase water solubility. Finally, the pre-
pared EFB with good chelating ability was utilized as a reusable
magnetic adsorbent to remove the heavy metal ions in the aqueous
solution.

3.2. Magnetic response analysis and the amount of nano-Fe3 O4

in the EFB

Fig. 2 shows the magnetic response of the EFB and yeast toward
the external magnet at the beginning (a) and after 100 s (b), respec-
tively. As shown in Fig. 2(a), the EFB suspension is on the left side,
whereas the yeast is on the right. Both EFB and yeast dispersed
uniformly in the aqueous solution at the beginning. In contrast,
Fig. 2(b) shows the EFB gathered on the cuvette wall 2.0 cm far
from the external magnet. Nevertheless, the suspension of yeast on
the right of the external magnet remained intact, proving that the
Fe3 O4 nanoparticles with superparamagnetism were firmly loaded
on the yeast and that the EFB was good at magnetic reclamation. In
addition, compared to the EDTAD-modified baker’s yeast reported
by Yu, the regeneration of EFB was cheaper and faster [18]. The
amount of nano-Fe3 O4 on EFB was measured using the GB1868-
1881 method and was calculated as 0.1003 g/g. This indicates that
the Fe3 O4 nanoparticles were on the baker’s yeast biomass surface,
which poses as a favorable superparamagnetism and is consistent
with the results from the magnetic response analysis.

3.3. Determination of active sites

The study of the proton binding active zone of the absorbent is of

great importance. Fig. 3 depicts a comparison of the titration curves Fig. 2. Magnetic response of EFB.
740 M. Xu et al. / Chemical Engineering Journal 168 (2011) 737–745

Fig. 3. Titration curves of the FB and EFB (FB/EFB: 3 g/L; titrant: 0.1 mol/L NaOH; Fig. 4. Effect of the pH value on the three metal ions adsorption by EFB (Pb2+ con-
reaction tempertaure: 25 ± 0.5 ◦ C; background, electrolyte: 0.1 mol/L NaCl). centration: 200 mg/L; Cd2+ and Ca2+ concentration: 300 mg/L; biomass dose: 1 g/L;
contact time: 30 min; reaction temperature: 10 ± 1 ◦ C).

should be associated with the difference in the amount of the func-

tional groups between the two samples, which most likely resulted the EFB [28,29]. The effect of pH on the adsorption capacity of EFB
from the EDTAD treatment. The pKa values of the organic functional for Ca2+ , Cd2+ , and Pb2+ in the pH ranging from 2 to 6 (for Pb2+
groups and the corresponding site densities cAi were calculated from 2 to 5.5) is presented in Fig. 4. The uptake of EFB for the
using the non-electrostatic model in the software ProtoFit [25,26]. three metal ions increased with an increase in pH. At low pH values,
By combining the microorganism cell wall structure and the com- the adsorption efficiency of EFB for the three metal ions was less
parisons of the previous reports, the carboxyl (pKa : 2.0–6.0), amino favorable because the protonation of the functional groups on EFB
(pKa : 9.0–11.0), hydroxyl (pKa : 8.0–12.0), and phosphoryl groups decreased the amount of available functional groups, coordinating
(pKa : 0.2–2.9, 5.6–7.2) are suggested to be mainly present in the the reaction with the metal ions. In addition, the positively charged
yeast biomass [27]. Analysis of the titration data indicated that a hydronium ions (H3 O+ ) competed with the metal cations for bind-
four-site model provided a good fit for both FB and EFB. The acidity ing on the surface functional groups (binding sites) on the EFB at
constants and site concentrations for each type of surface functional lower pH values [3]. As the deprotonation degree of the functional
groups on FB and EFB are summarized in Table 1. The total concen- groups increased simultaneously with the increase in pH, the coor-
tration of the carboxyl and amine groups on EFB was higher than dination ability between the functional groups on the EFB and the
that on FB. Apparently, the EDTAD treatment enhanced the number metal ions in the solution were enhanced. The maximum uptakes
of carboxyl and amine groups on the surface of FB. Furthermore, (88.16 mg/g for Pb2+ , 40.70 mg/g for Cd2+ , and 27.19 mg/g for Ca2+ )
the repeated vacuum freeze-drying as well as the dissolution of were observed at approximately pH 5.5 and pH 6.0, respectively.
DMF to yeast cell wall (mannan) resulted in the destruction of the Compared with the ethanol treated baker’s yeast biomass [30] and
outer structure layer of the yeast cell wall, exposing more phos- the cystine-modified baker’s yeast biomass [31], the adsorption
phoryl groups in the cell wall. As a consequence, the amount of capacity of Pb2+ , Cd2+ by EFB is higher.
phosphoryl on EFB was higher than that on FB. Moreover, the tem-
perature will affect the extent of the non-protonated functional 3.5. Effect of contact time and adsorption kinetics
group on the surface of EFB. The amount of non-protonated func-
tional group on EFB increased along with the rise in the reaction Adsorption equilibrium time is one of the parameters for eco-
temperature. nomical waste water treatment in practical applications. Different
contact times (from 5 to 120 min) were tested at pH values (5.5
3.4. Effects of pH on adsorption for Ca2+ and 6.0 for Cd2+ and Pb2+ ) at 10 ◦ C using 0.1 g of the EFB.
Fig. 5 presents the adsorption kinetics of Ca2+ , Cd2+ , and Pb2+ on
The pH of the aqueous solution plays an important role with EFB. Lead, cadmium, and calcium uptake increased rapidly, and
respect to adsorption because it influences not only the metal almost 90% adsorption capacity was attained in about 5 min. The
species distribution in solution but also the surface properties of quick and efficient adsorption process illustrates that the adsorp-
tion of metal ions almost occurred on the surface of EFB, as well
Table 1 as the abundant availability of binding sites present on the surface
Concentration and acidity constants of the surface functional groups of FB and EFB. of the baker’s yeast. The biosorption equilibrium was established
within 30 min, which was shorter than that in other biosorbents
Adsorbent Functional group pKa values Concentration of
obtained functional groups [32]. Therefore, the optimum contact time was selected as 30 min
(mmol/g) for further experiments.
FB Carboxyl 4.07–4.15 0.27 ± 0.05 To analyze the adsorption kinetics of the metal ions onto the
Phosphoryl 6.34–6.80 0.21 ± 0.04 EFB, the pseudo-first-order, kinetics pseudo-second-order kinetics,
Amine 8.16–9.04 0.35 ± 0.02 and intraparticle diffusion models were tested using the obtained
Hydroxyl 9.84–10.34 0.79 ± 0.03 experimental data and rate constants. The pseudo-first-order
EFB Carboxyl 4.51–4.83 0.87 ± 0.07 kinetics, pseudo-second-order kinetics, and intraparticle diffusion
Phosphoryl 6.23–6.77 0.76 ± 0.06 equations are given in the following forms [33]:
Amine 8.28–9.12 0.72 ± 0.02
Hydroxyl 9.35–10.45 0.79 ± 0.04 pseudo-first-order equation : ln(qe − qt ) = −k1 t + ln qe (4)
 M. Xu et al. / Chemical Engineering Journal 168 (2011) 737–745
Fig. 5. Adsorption of metal ions onto EFB as a function of time (300 mg/L for
Ca2+ /Cd2+ at pH 6 and 200 mg/L for Pb2+ at pH 5.5. Biomass dose: 1 g/L; reaction
temperature: 10 ± 1 ◦ C).
t 1 t
pseudo-second-order equation : = + (5)
qt k2 q2e bqe
intraparticle diffusion equation : qt = ki t 0.5 + I (6)
where k1 is the Lagergren adsorption rate constant (h−1 ),
k2 is the
pseudo-second-order adsorption rate constant [g/(mg h)], qe and qt
are the amounts of mental ions absorbed (mg/g) at equilibrium and
time t, ki is the intraparticle diffusion rate constant (mg/g min−0.5 ),
and I (mg/g) is a constant that provides information on the thick-
ness of the boundary layer. The estimated parameters are shown
in Table 2. The values of the calculated adsorption capacity (qe,cal )
were close to the experimental values (qe,exp ) for the second-order
and intraparticle diffusion kinetics compared with the first-order
kinetics equation. However, the correlation coefficient values (R2 )
for intraparticle diffusion kinetics were much lower than those for
the pseudo-second-order kinetics. These results indicate that the
adsorption process according to the pseudo-second-order kinetics
model is better than the two other models. Thus, the chemisorption
occurred in the adsorption process.
3.6. Effect of initial concentration on adsorption
Studies on the adsorption isotherm are prerequisites to under-
standing the adsorbate–adsorbent interaction and to optimize the
use of the adsorption. Fig. 6 shows the effect of the three metal ions
concentrations (20–200 mg/L for Pb2+ at pH 5.5 and 20–300 mg/L
for Cd2+ /Ca2+ at pH 6.0) on the adsorption capacity at 10 ◦ C. The
adsorption capacity of EFB for the three metal ions increased along
with the increase in the initial metal ions concentrations, ultimately
attaining a saturated value. This is attributed to the increase in the
driving force among the metal ions at a high initial concentration
that enhanced the effective collision probability between the metal
ions and EFB. The almost stationary adsorption capacity against
any further increase in the metal ions concentration ions could be
explained by the lack of sufficient adsorption sites to accommo-
date more metal ions available in the solution. Similar findings have
also been reported by other researchers [34,35]. These results sug-
gest that the available functional groups on EFB are the limiting
factors in metal ions adsorption. These experimental data were fit-
ted by the Langmuir and Freundlich adsorption isotherm models.
The parameters and correlation coefficients of the two adsorption
isotherms evaluated at 10 ◦ C are presented in Table 3. The linear
forms of the Langmuir and Freundlich equations are expressed as
741
Fig. 6. Effect of the initial concentration on adsorption (Ca2+ /Cd2+ at pH 6 and Pb2+ at
pH 5.5; biomass dose: 1 g/L; contact time: 30 min; reaction temperature: 10 ± 1 ◦ C).
follows:
Ce Ce 1
Langmuir equation : = + (7)
qe qm bqm
ln Ce
Freundich equation : ln qe = + ln KF (8)
n
where qe and qm are the equilibrium adsorption capacity and the
maximum adsorption capacity of EFB (mg/g), respectively, ce is the
equilibrium metal ion concentration in the solution (mg/L), b is
the Langmuir adsorption equilibrium constant (L/mg) related to
the free energy and affinity of adsorption, kF is a constant (L1/n
mg1/n−1 /g) related to the adsorption capacity, and 1/n is an empiri-
cal parameter related to the adsorption intensity, which varies with
the heterogeneity of the materials. Compared with the Langmuir
and Freundlich correlation coefficients (R2 ) presented in Table 3,
the adsorption process of the EFB for the three metal ions was
more consistent with the Langmuir isotherm equation model. Sim-
ilar findings have also been reported by other researchers [30,36]. A
linearized plot of ce /qe versus ce was obtained from the models (not
shown), illustrating that a monolayer adsorption process occurred
on the surface of EFB. Moreover, the qm of the FB obtained at 10 ◦ C in
the same conditions as that of EFB for Pb2+ , Cd2+ , and Ca2+ is 21.78,
13.02, and 5.38 mg/g, respectively, which are lower than those of
the EFB. The results further confirmed that the FB was successfully
treated by EDTAD. The Langmuir constant b can be used to deter-
mine the suitability of the adsorbent for the sorbate using the Hall
separation factor (RL ) as follows [37]:
1
RL = (9)
1 + bCm
where Cm is the highest initial metal ion concentration (mg/L). RL
is the constant separation factor (dimensionless) and can be used
for the interpretation of the sorption type as follows [38]:
RL > 1 unfavorable
RL < 0 unfavorable
RL = 1 favorable (linear)
0 < RL < 1 favorable
RL = 0 irreversible
As shown in Table 3, the value of RL for Pb2+ , Cd2+ , and Ca2+
is 0.0095, 0.0077, and 0.0154, respectively, indicating a favorable
adsorption of the three metal ions on the EFB.
742 M. Xu et al. / Chemical Engineering Journal 168 (2011) 737–745

Table 2
Comparison of the pseudo-first-order, pseudo-second-order, and intraparticle diffusion adsorption rate constants and calculated and experimental qe values.

Metal ions qe exp (mg/g) Pseudo-first-order kinetic model Pseudo-second-order kinetic model Intraparticle diffusion kinetic model
−1 −1
k1 (min ) R2
qe cal (mg/g) k2 (g/mg min ) R 2
qe cal (mg/g) ki (mg/g min−0.5 ) R2 qe cal (mg/g)

Ca2+ 27.30 −0.0125 0.5898 1.3751 −0.0156 0.9973 22.11 −0.4243 0.5838 25.45
Cd2+ 41.55 0.0042 0.0668 3.8421 −0.0535 0.99899 38.23 0.3346 0.1594 38.23
Pb2+ 88.16 −0.1082 0.3512 1.6912 −0.0152 0.9998 83.82 −0.5441 0.4550 85.65

Table 3
Langmuir and Freundlich isotherm constants and correlation coefficients for adsorption.

Metal ions Langmuir constant Freundlich constant

qm (mg/g) b (L/mg) R2 RL 1/n kF (L1/n mg1/n−1 /g) R2

Ca2+ 26.50 0.3468 0.9993 0.0095 0.1644 16.02 0.9273

Cd2+ 41.00 0.4280 0.9995 0.0077 0.1578 18.77 0.9255
Pb2+ 89.21 0.3192 0.9902 0.0154 0.1458 43.91 0.9700

3.7. Effect of temperature on adsorption Table 4

pKw at different temperatures.

At different temperature ranges (10–60 ◦ C), 0.1 g of EFB was T (◦ C) 10 20 30 40 50 60

added to a 100.0 mL solution with different metal ions, namely,
300 mg/L for Ca2+ /Cd2+ at pH 6 and 200 mg/L for Pb2+ at pH 5.5.
As shown in Fig. 7, the adsorption capacity of the EFB for the
three metal ions was obviously affected by temperature. The uptake
increased with a rise in the reaction temperature from 10 to 30 ◦ C,
and the maximum adsorption capacities of Pb2+ , Cd2+ , and Ca2+
were 99.26, 48.70, and 33.46 mg/g at 30 ◦ C, respectively. First, at a
low temperature (10–30 ◦ C), the amount of non-protonated func-
tional group on EFB increased due to the increasing ionization
constant of protonated functional groups along with the rise in the
reaction temperature. Second, the mobility of metal ions increased
as the temperature increased. However, the adsorption capacity of
EFB decreased with the increasing reaction temperature from 30
to 60 ◦ C. The increasing temperature impelled the hydrolysis of the
ester bond between the EDTA molecules due to the endothermic
property of the hydrolysis reaction [39]. The ester bonds were dis-
rupted, and the EDTA molecules were subsequently released from
the surface of the EFB. Therefore, the hydrolysis reaction of the ester
bond restricted the adsorption of EFB for the metal ions. Moreover,
because the increase in the dissociation constant of water increased
simultaneously with the rapid increase in temperature (shown in
Table 4) [40], the concentration of hydronium (H3 O+ ) and hydroxyl
(OH− ) increased in the solution. The increasing concentration of
hydrogen ions resulted in the protonation of the functional groups
pKw 14.528 14.163 13.836 13.542 13.275 13.034
on the EFB, leading to the decrease in the adsorption capacity of EFB
for the metal ions. The metal species distribution in the solution
changed by the increasing hydroxyl concentration in the solution
directly caused the adsorption capacity of the EFB for metal ions to
decrease. Therefore, from 10 to 30 ◦ C, the increasing uptake of EFB
for the three metal ions with the rise in the reaction temperature
was due to the increasing amount of non-protonated functional
groups and the mobility of metal ions. On the contrary, the decreas-
ing adsorption capacity of the EFB for metal ions was primarily
affected by the increase in the hydrolysis reaction and the dissoci-
ation constant of water between 30 and 60 ◦ C.
3.8. Effect of ternary metals ions mixtures on adsorption
The results of the adsorption of the single Pb2+ , Cd2+ , Ca2+ , and
the ternary Pb2+ –Cd2+ –Ca2+ system (100 mg/L for Pb2+ , Cd2+ , Ca2+
at pH 5.5) on EFB/FB are presented in Table 5, respectively. The
adsorption capacity of the EFB in the ternary metal ions system was
lower than that in the single metal ion system, of which the adsorp-
tion capacity of EFB for Pb2+ decreased less than that of Cd2+ and
Ca2+ . Moreover, for the FB, the uptake of the three metal ions was
lower than that of the EFB under the same condition, the adsorption
capacity in the coexisting metals condition apparently decreased
compared with the single metal condition. This shows that the EFB
possessed a better selective adsorption capacity for the metal ions
than the FB. Given that the EDTA molecules were successfully intro-
duced to the surface of FB by EDTAD treatment, and the stability
constant of Pb-EDTA was much higher than those of Cd-EDTA and
Ca-EDTA [41], the competitive ability of lead(II) ions for the limited
adsorption sites on EFB was stronger than that of the other two
ions. Thus, the massive Pb2+ ions were adsorbed by the EFB.

Table 5
Effect of the different coexisting cations on the adsorption of metal ions by EFB
and FB (Pb2+ , Cd2+ and Ca2+ concentration: 100 mg/L; contact time: 30 min; reaction
temperature: 10 ± 1 ◦ C; pH 5.5).

Adsorbent Uptake in single metal Uptake in ternary metal

ions solution (mg/g) ions solution (mg/g)

Pb2+ Cd2+ Ca2+ Pb2+ Cd2+ Ca2+

FB 13.53 8.24 4.41 7.31 3.55 2.09

Fig. 7. Effect of temperature on adsorption(300 mg/L for Ca2+ /Cd2+ at pH 6 and
EFB 66.89 38.66 25.38 55.45 23.37 3.52
200 mg/L for Pb2+ at pH 5.5. biomass dose: 1 g/L).
 M. Xu et al. / Chemical Engineering Journal 168 (2011) 737–745
Fig. 8. Comparison of IR spectra of FB and EFB with/without metal ions (a: FB; b:
EFB; c: EFB with Pb2+ ; d: EFB with Cd2+ ; e: EFB with Ca2+ ).
3.9. Desorption and regeneration studies
The regeneration of the adsorbent is a significant factor in
assessing its potential for commercial application. The adsorp-
tion/desorption (A/D) cycles of Pb2+ , Cd2+ , and Ca2+ were repeated
thrice using different eluents of 0.1 mol/L HCl and 0.1 mol/L DTPA,
respectively. Table 6 shows the minimal change in the uptake and
metal recovery efficiency of EFB for the three metal ions after three
consecutive adsorption/desorption cycles, indicating that the EFB
is an effective and reusable adsorbent and that HCl/DTPA are good
eluents of metal ions. In addition, the metal recovery efficiency
with the eluent HCl was higher compared with the eluent DTPA
after the same cycle of adsorption/desorption. This was due to the
stability of the metal ion-ligands between the calcium, lead, and
cadmium ions and the EDTA molecules introduced to baker’s yeast
biomass, which was significantly affected by the concentration of
H+ in solution. Thus, the complexation of the metal ion with EDTA
was disrupted by HCl, and the metal ions were released from the
surface of EFB.
3.10. FTIR analysis
Infrared (IR) spectra is important in identifying the organic func-
tional groups of EFB as well as deducing its treated and adsorption
mechanism [42,43]. To confirm the EDTA bond in the FB and the
adsorption mechanism of the EFB on the metal ions (Pb2+ , Cd2+ and
Ca2+ ), the IR spectra for FB, EFB and metal ions-loaded EFB were
investigated, respectively. As shown in Fig. 8(a), the FB had a very
complex structure and a number of absorption peaks. The broad
strong band at 3445 cm−1 corresponded with the overlapping
of –NH/–OH stretching. The peaks at 1645 cm−1 and 1566 cm−1
showed the asymmetric stretching vibration band of C–O from
the amide I band and the bending band of N–H from the amide
II band, respectively. They are the characteristic absorption peaks
of protein. The band at 1412 cm−1 belonged to the symmetrical
stretching band of carboxyl (CO–O), the weak peaks at 1153 cm−1
came from the coupling of the stretching band of C–N, and the
absorption peaks at 1069 cm−1 and 955 cm−1 were attributed to
the P–O–C and glycitols (C–OH), respectively [18,44]. The vibrating
absorption peaks at 868 cm−1 was the stretching band of N–H from
amide III. Moreover, the significant peaks at 540 cm−1 with respect
to the characteristic stretching band of Fe–O further confirmed the
loading of nano-Fe3 O4 onto the biomass surface. Thus, the main
organic functional groups on FB were carboxyl, amino/hydroxyl,
phosphoryl, and others.
743
Fig. 9. Comparison of the zeta potential of FB/EFB with/without metal ions (FB/EFB
dose: 1 g/L; concentration: 300 mg/L for Ca2+ /Cd2+ , 200 mg/L for Pb2+ ; contact time:
30 min; pH 5).
As shown in Fig. 8(b), there are apparent differences between
the EFB and FB spectra. For EFB, not only did the decrease in
the absorption peak strength of –NH/–OH stretching vibration
occur, but the broad overlapping peak also moved from 3445 to
3447 cm−1 to a high frequency direction. It is possible that some of
the hydroxyl groups on the surface of the baker’s yeast biomass par-
ticipated in the esterification process. A significant absorption peak
at 1747 cm−1 representing the carbonyl group appeared, and the
symmetrical stretching band of the carboxyl (CO–O) group moved
from 1412 cm−1 to 1418 cm−1 in the IR spectrum of EFB, indicat-
ing that a certain amount of the carboxyl groups was introduced to
the surface of EFB by treatment of EDTAD. In addition, the stretch-
ing band of C–N moved from 1153 to 1161 cm−1 , and the peak
strength evidently increased. The bending band of N–H from the
amide II band moved from 1566 to 1570 cm−1 . All these mean that
the hydroxyl groups on the surface of the baker’s yeast participated
in the esterification reaction with the carboxyl groups in the EDTA
molecules. Hence, treatment of EDTAD for FB resulted in an increase
in the amount of the carboxyl and amino groups on the surface of
EFB.
Fig. 8(c–e) shows the changes in the IR spectrum of the three
metal ions’ (Pb2+ , Cd2+ and Ca2+ ) biosorption by EFB. After adsorp-
tion, the peaks at 3447 cm−1 attributed to the –NH/–OH absorbance
shifted to different peaks at 3448, 3427 and 3433 cm−1 ; the charac-
teristic peaks representing carbonyl also moved 12, 10 and 9 cm−1 ,
respectively. Moreover, the characteristic absorbance peaks indi-
cating amide I and II also exhibited the same changes. Among
them, the peaks at 1651 cm−1 , representing amide I after adsorp-
tion moved 4 cm−1 to a low wavenumber direction, and 1570
representing amide II after adsorption moved 8 cm−1 to a high
wavenumber direction. Hence, these changes indicate that the car-
boxyl, amino/hydroxyl groups on the EFB were mainly involved in
the adsorption process of the three metal ions in various degrees.
3.11. Zeta potential analysis
The zeta potential showing the surface charge of EFB and FB
was used to study the behavior of the three metal ions adsorbed by
the adsorbent. Large quantities of negatively charged organic func-
tional groups, such as carboxyl, amino, phosphoryl, and so on, are
present on the baker’s yeast cell wall containing amylase, protein,
lipid, chitin, and others [45,46]. Hence, yeast biomass has gener-
ally some attraction with cations in a solution under a certain pH
range. The zeta potentials of EFB and FB before and after adsorbing
the three metal ions shown in Fig. 9 were measured at pH 5. The zeta
potentials of EFB and FB had negative charges (−15.78, −3.26 mV)
744 M. Xu et al. / Chemical Engineering Journal 168 (2011) 737–745

Table 6
Adsorption and desorption (recovery) of metal ions on EFB (Pb2+ concentration: 200 mg/L, pH 5.5; Cd2+ and Ca2+ concentration: 300 mg/L, pH 6.0; EFB dose: 1 g/L; contact
time: 30 min; reaction temperature: 10 ± 1 ◦ C).

Metal ions A/D cycle HCl ETPA

Uptake Metal recovery Uptake Metal recovery

(mg/g) efficiency (%) (mg/g) efficiency (%)

Ca2+ 1 27.19 90.03 27.19 85.46

2 23.90 87.93 23.07 83.70
3 20.16 85.20 18.78 82.17

Cd2+ 1 40.72 89.64 40.72 81.71

2 34.69 85.55 32.92 75.33
3 32.55 80.27 25.20 70.17

Pb2+ 1 88.16 85.58 88.16 78.21

2 72.99 82.82 60.66 70.57
3 71.77 81.47 58.47 68.89

with the following order and strength of charge: EFB > FB. This indi-
cates that the EDTAD treatment enhanced the amount of carboxyl
and amino groups on the surface of FB. Moreover, the repeated
vacuum freeze-drying as well as the dissolution of DMF to the
yeast cell wall resulted in the destruction of the outer structure
layer of the yeast cell wall; the negatively charged intracellu-
lar functional groups (i.e., phosphoryl group) also became more
exposed. More negatively charged functional groups likely caused
the increase in the electrostatic attraction between the EFB and the
metal cations. Furthermore, the zeta potential of EFB changed to
−8.68, −10.45, −11.73 mV, whereas that of the FB changed to −1.85,
−2.45, −2.83 mV, respectively, after they adsorbed Pb2+ , Cd2+ , and
Ca2+ . Perhaps it was due to the positive charge of the metal ions
neutralizing the negative charges of the EFB and the FB. Based
on the charge changes, the electrostatic interaction appears to be
advantageous in adsorbing metal cations.
4. Conclusions
FB was prepared using glutaraldehyde as a cross link agent and
was chemically treated with EDTAD. EFB was investigated by a
number of techniques. Magnetic response analysis showed that the
EFB had superparamagnetism ability as an external magnet. The
results of IR and potentiometric titration indicated that the amount
of the carboxyl and amino groups were higher on the surface of
the EFB than that of the FB. The adsorption properties of EFB for
Pb2+ , Cd2+ , and Ca2+ were also investigated in our study. The maxi-
mum adsorption capacity (88.16 mg/g for Pb2+ at pH 5.5, 40.72 mg/g
for Cd2+ , and 27.19 mg/g for Ca2+ at pH 6.0) was obtained at 10 ◦ C.
The adsorption process was described well by the pseudo-second-
order kinetics model and the Langmuir isotherm equation. The zeta
potential of EFB and FB changed before and after the adsorption
of the metal ions. The regeneration of the EFB was achieved using
0.1 mol/L HCl and 0.1 mol/L DTPA with over 68% metal recovery effi-
ciency. Thus, EFB is a potentially recyclable, effective, and selective
adsorbent for heavy metal disposals.
Acknowledgments
This research was supported by the Key Basic Research Program
of the Sichuan Provincial Education Commission, PR China (Grant
Nos. 10ZB034, 07ZA063, 2005A014) and the Basic Research Pro-
gram of the Science & Technology Department of Sichuan Province
(Grant No. 2006J13-039).
References
[1] B. Kiran, A. Kaushik, C.P. Kaushik, Response surface methodological approach
for optimizing removal of Cr (VI) from aqueous solution using immobilized
cyanobacterium, Chem. Eng. J. 126 (2007) 147–153.
[2] D. Mohan, C.U. Pittman Jr., Activated carbons and low cost adsorbents for reme-
diation of tri- and hexavalent chromium from water, J. Hazard. Mater. 137
(2006) 762–811.
[3] P.X. Sheng, Y.P. Ting, J.P. Chen, L. Hong, Sorption of lead, copper, cadmium,
zinc, and nickel by marine algal biomass: characterization of biosorptive
capacity and investigation of mechanisms, J. Colloid Interface Sci. 275 (2004)
131–141.
[4] P.B. Kelter, J. Grundman, D.S. Hage, J.D. Carr, C.M. Castro-Acuña, A discussion of
water pollution in the United States and Mexico; with high school laboratory
activities for the analysis of lead, atrazine, and nitrate, J. Chem. Educ. 74 (1997)
1413–1421.
[5] World Health Organization, Recommendations Guidelines for Drinking
Water Quality, vol. 1, 2nd ed., World Health Organization (WHO), Geneva,
Switzerland, 1993.
[6] S. Babel, T.A. Kurniawan, Low-cost adsorbents for heavy metals uptake from
contaminated water: a review, J. Hazard. Mater. B97 (2003) 219–243.
[7] P. Marques, M.F. Rosa, H.M. Pinheiro, pH Effects on the removal of Cu2+ , Cd2+ and
Pb2+ from aqueous solution by waste brewery biomass, Biotechnol. Bioprocess
Eng. 23 (2000) 135–141.
[8] K.M. Hong, M.S. Kim, J.G. Chung, Adsorption characteristics of Ni(II) on c-type
alumina particles and its determination of overall adsorption rate by a differ-
ential bed reactor, Chemosphere 54 (2004) 927–934.
[9] H. Yavuz, A. Denizli, H. Güngünes, M. Safarikova, I. Safarik, Biosorption of
mercury on magnetically modified yeast cells, Sep. Purif. Technol. 52 (2006)
253–260.
[10] V.K. Gupta, I. Ali, Removal of lead and chromium from wastewater using
bagasse flyash – a sugar industry waste, J. Colloid Interface Sci. 271 (2004)
321–328.
[11] D. Sud, G. Mahajan, M.P. Kaur, Agricultural waste material as potential adsor-
bent for sequestering heavy metal ions for aqueous solutions – a review,
Bioresour. Technol. 99 (2008) 6017–6027.
[12] K.K. Krishnani, S. Ayyappan, Heavy metals remediation of water using plants
and lignocellulosic agrowastes, Rev. Environ. Contam. Toxicol. 188 (2006)
59–84.
[13] U.K. Garg, M.P. Kaur, V.K. Garg, D. Sud, Removal of hexavalent chromium from
aqueous solution by agricultural waste biomass, J. Hazard. Mater. 140 (2007)
60–68.
[14] Y.S. Zhang, W.G. Liu, M. Xu, F. Zheng, M.J. Zhao, Study of the mechanisms of
Cu2+ biosorption by ethanol/caustic-pretreated baker’s yeast biomass, J. Haz-
ard. Mater. 178 (2010) 1085–1093.
[15] D.M. Borrok, J.B. Fein, The impact of ionic strength on the adsorption of pro-
tons, Pb, Cd, and Sr onto the surfaces of Gram negative bacteria: testing
non-electrostatic, diffuse, and triple-layer models, J. Colloid Interface Sci. 286
(2005) 110–126.
[16] R. Han, H. Li, Y. Li, J. Zhang, H. Xiao, J. Shi, Biosorption of copper and lead ions
by waste beer yeast, J. Hazard. Mater. B137 (2006) 1569–1576.
[17] J.L. Wang, Biosorption of copper(II) by chemically modified biomass of Saccha-
romyces cerevisiae, Process Biochem. 37 (2002) 847–850.
[18] J.X. Yu, M. Tong, X.M. Sun, B.H. Li, Enhanced and selective adsorption of Pb2+
and Cu2+ by EDTAD-modified biomass of baker’s yeast, Bioresour. Technol. 99
(2008) 2588–2593.
[19] S.G. Wang, Y.H. Li, X.Y. Gong, Z. Luan, C. Xu, Surface characteristics of modified
carbon nanotubes and its application in lead adsorption from aqueous solution,
Chin. Sci. Bull. 48 (2003) 441–443.
[20] J.Y. Wei, Y.J. Zhang, F. Tan, H.L. Liu, J.L. Wang, Y. Cai, X.H. Qian, A
novel Desalting method on target for MALDI-TOF-MS analysis based on
Fe3 O4 /TiO2 nanoparticles, Chin. J. Anal. Chem. 35 (2007) 1–7.
[21] W.J. Chen, P.J. Tsai, C.C. Yu, Functional Fe3 O4 /TiO2 core/shell magnetic nanopar-
ticles as photokilling agents for pathogenic bacteria, Small 4 (2008) 485–491.
[22] X.X. Wang, S. Huang, Z. Shan, Preparation of Fe3 O4 @Au nano-composites by
self-assembly technique for immobilization of glucose oxidase, Chin. Sci. Bull.
54 (2009) 1176–1181.
[23] A. Capretta, R.B. Maharajh, R.A. Bell, Synthesis and characterization of
cyclomaltoheptaose-based metal chelants as probes for intestinal permeabil-
ity, Carbohydr. Res. 267 (1995) 49–63.
 M. Xu et al. / Chemical Engineering Journal 168 (2011) 737–745
[24] S.Q. Memon, N. Memon, S.W. Shah, M.Y. Khuhawar, M.I. Bhanger, Sawdust – a
green and economical sorbent for the removal of cadmium(II) ions, J. Hazard.
Mater. 139 (2007) 116–121.
[25] B.F. Turner, J.B. Fein, Protofit: a program for determining surface protonation
constants from titration data, Comput. Geosci. 32 (2006) 1344–1356.
[26] A. Barkleit, H. Moll, G. Bernhard, Interaction of uranium(VI) with lipopolysac-
charide, J. Chem. Soc., Dalton Trans. 21 (2008) 2879–2886.
[27] Y. Nathan, L.G. Benning, V.R. Phoenix, F.G. Ferris, Characterization of met-
alcyanobacteria sorption reactions: a combined macroscopic and infrared
spectroscopic investigation, Environ. Sci. Technol. 38 (2004) 775–782.
[28] H. Bessbousse, T. Rhlaloub, J.-F. Verchère, L. Lebruna, Mercury removal from
wastewater using a poly(vinylalcohol)/poly(vinylimidazole) complexing mem-
brane, Chem. Eng. J. 164 (2010) 37–48.
[29] P.A. Marques, H.M. Pinheiro, J.A. Teixeira, M.F. Rosa, Removal efficiency of Cu2+ ,
Cd2+ , Pb2+ by waste brewery biomass: pH and cation association effects, Desali-
nation 124 (1999) 137–144.
[30] Y. Göksungur, S. Üren, U. Guvenç, Biosorption of cadmium and lead ions by
ethanol treated waste baker’s yeast biomass, Bioresour. Technol. 96 (2005)
103–109.
[31] J.X. Yu, M. Tong, X.M. Sun, B.H. Li, Cystine-modified biomass for Cd(II) and Pb(II)
biosorption, J. Hazard. Mater. 143 (2007) 277–284.
[32] S. Veli, B. Pekey, Removal of copper from aqueous solution by ion exchange
resins, Fresenius Environ. Bull. 13 (2004) 244–250.
[33] Y.S. Ho, G. McKay, Pseudo-second order model for sorption processes, Process
Biochem. 34 (1999) 451–465.
[34] R. Nadeem, M.H. Nasir, M.S. Hanif, Pb (II) sorption by acidically modified Cicer
arientinum biomass, Chem. Eng. J. 150 (2009) 40–48.
[35] L.Y. Wang, L.Q. Yang, Y.F. Li, Y. Zhang, X.J. Ma, Z.F. Ye, Study on adsorption
mechanism of Pb(II) and Cu(II) in aqueous solution using PS-EDTA resin, Chem.
Eng. J. 163 (2010) 364–372.
745
[36] J.X. Yu, M. Tong, X.M. Sun, B.H. Li, A simple method to prepare poly(amic acid)-
modified biomass for enhancement of lead and cadmium adsorption, Biochem.
Eng. J. 33 (2007) 126–133.
[37] R. Nadeem, M.A. Hanif, F. Shaheen, S. Perveen, M.N. Zafar, T. Iqbal, Physical
and chemical modification of distillery sludge for Pb(II) biosorption, J. Hazard.
Mater. 150 (2008) 335–342.
[38] S. Dahiya, R.M. Tripathi, A.G. Hegde, Biosorption of heavy metals and radionu-
clide from aqueous solutions by pre-treated arca shell biomass, J. Hazard. Mater.
150 (2008) 376–386.
[39] N. Olaru, A. Andriescu, L. Olaru, On the hydrolysis of cellulose acetate in
toluene/acetic acid/water system, Eur. Polym. J. 37 (2001) 865–867.
[40] W.L. Marshall, E.U. Franck, J. Phys. Chem. Ref. Data 10 (1981) 295.
[41] J.A. Dean, Lange’s Handbook of Chemistry, 5th ed., McGraw-Hill Inc., 1998.
[42] E. Kiefer, L. Sigg, P. Schosseler, Chemical and spectroscopic characterization of
algae surfaces, Environ. Sci. Technol. 31 (1997) 759–764.
[43] M. Bansala, U. Garg, D. Singh, V.K. Garg, Removal of Cr(VI) from aqueous solution
using pre-consumer processing agricultural waste: a case study of rice husk, J.
Hazard. Mater. 162 (2009) 312–320.
[44] K. Nakanish, Infrared Absorption Spectroscopy – Practical, Holden-Day Inc., San
Francisco, 1962, pp. 38–47.
[45] Y.S. Zhang, R.G. Wang, X.X. Dai, S.Z. Lei, D.M. Tong, Study on comparison of
the cell morphology of baker’s yeast biomass modified by chemical treatment
and their characteristics of Cu2+ adsorption, Acta Sci. Circumstantiae 28 (2008)
897–901 (in Chinese).
[46] D. Brady, A.D. Stoll, L. Starke, J.R. Duncan, Chemical and enzymatic extraction
of heavy-metal binding polymers from isolated cell walls of Saccharomyces
cerevisiae, Biotechnol. Bioeng. 44 (1994) 297–302.