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Removal of Copper and Lead Ions from Aqueous Solution Using Brewer Yeast
as Biosorbent

Article  in  Revista de Chimie -Bucharest- Original Edition- · July 2016

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 Removal of Copper and Lead Ions from Aqueous Solution
Using Brewer Yeast as Biosorbent

ANDREEA STANILA1, TANIA MIHAIESCU2*, CARMEN SOCACIU1, ZORITA DIACONEASA1

1
University of Agricultural Sciences and Veterinary Medicine, Faculty of Food Science and Technology, 400372, 3-5 Manastur St.,
Cluj-Napoca, Romania
2
University of Agricultural Sciences and Veterinary Medicine, Faculty of Agriculture, 400372, 3-5 Manastur St., Cluj-Napoca,
Romania

Biosorption is an effective procedure of heavy metal ions removal from wastewater. Brewer yeast, a byproduct
of brewing industry could be a low cost and promising adsorbent for copper and lead ions from aqueous
solutions. In this work the removal of these ions from water solutions by brewer yeast was performed as
function of key parameters such pH, contact time, yeast dose and initial metal ions concentrations. Results
demonstrate that the highest equilibrium adsorption capacity was reached at the pH=6 for both metal ions.
It was found that the adsorptive capacity qe of both copper and lead ions on brewer yeast increase with the
increasing of the contact time until the equilibrium stage was obtained. The biosorption process was rapid
in the first 15 min. and the equilibrium was reached after 60 min. Results showed that the percent of metal
removal increase as yeast dosage increase up to 1 g/100 mL solution for metal ions. The increase in metal
uptake with yeast dosage confirms the increase in binding sites for metal ions. The metal uptake decrease
with the increasing of yeast dosage over 1 g/100 mL due to aggregation of the yeast. Biosorption experiment
was carried out at different initial concentrations of Cu(II) and Pb(II) ions, ranged between 1 mg L-1 to 10 mg
L-1. The removal efficiency increase with the increasing of metal ions concentrations until 7 mg L-1 for Pb(II)
ions and 8 mg L-1 for copper ions. The sorption equilibrium data were analyzed using the Langmuir adsorption
isotherms. The data obtained represent a favorable adsorption in the case of adsorption of Cu(II) and Pb(II)
ions. The maximum loading capacities of beer yeast was obtained as 20.6 mg g-1 for Cu(II) ions and 22.9 mg
g-1 for Pb(II) ions, so the ability of lead adsorption is higher than copper ions.
Keywords: biosorption, brewer yeast, metal ions removal, Saccharomyces cerevisae

Heavy metals are discharged from various industries
and pose serious environmental and human health
problems. Biosorption represent the removal of metal
species compounds from aqueous solution by biological
materials such as bacteria, fungi, algae and yeast [1].
Biosorption exploits the ability of microbial biomass to
sequester heavy metal ions from aqueous solution by
physico-chemical mechanisms [2, 3]. Charged groups
such carboxylate, amines and hydroxyl present in the
biopolymers of biomass cell walls are believed to be
responsible for the sequestration of metal ions [4].
A number of literature data have proved that S. cerevisiae
can remove toxic metals, recover precious metals and
clean radionuclides from aqueous solutions to various
extents [6-12].
The biosorption of heavy metals using microorganisms
like S. cerevisae is affected by several factors. These factors
include the specific surface properties of the biosorbent
and the physico-chemical parameters of the solution such
as temperature, pH, initial metal ion concentration and
biomass concentration [13]. The pretreatment and killing
of biomass either by physical or chemical treatments or
crosslinking are known to improve the biosorption capacity
of biomass. Carboxylate and amine in yeast play an
important role in metal biosorption [14].
Copper and lead ions were chosen for this study with
regard to their wide use in industry and due to their potential
pollution impact. Copper, which is a very widely used metal,
like others heavy metal could be potentially toxic for the
living organisms. Lead can contaminate the environment
from anthropogenic sources as well as natural
geochemical processes. It can accumulate along the food
chain and is not amenable to biological degradation [15].
* email: tmihaescu@yahoo.com
1276 http://www.revistadechimie.ro
The aim of this study was to use brewer’s yeast as
biosorbent for copper and lead ions in aqueous solution.
The objective of this work was to study the influence of the
uptake of Cu(II) and Pb(II) by brewer’s yeast in different
adsorptive conditions.
Experimental part
Biomass and chemicals preparation
Saccharomyces cerevisae biomass was supplied as a
lyophilized by-product from industrial ethanol production.
The waste biomass as washed with deionized water by
stirring followed by centrifugation at 3000 rpm for 20
minutes. The supernatant was discarded and the pellet
was reslurried in deionized water. The procedure was
repeated for three times until the supernatant was clear.
The chemicals used for this study were analytical grades
of Pb(NO3)2, CuSO4·5H2O, HCl, NaOH, 4-(2-pyridylazo)
resorcinol, purchased from Merck (Darmstadt, Germany).
Stock solution of 100 mg L-1 Cu(II) and Pb(II) were
prepared from the above metal salts dissolved in deionized
water. The stock solutions were diluted as required to obtain
working solution. The pH of the working solutions was
adjusted by addition of HCl 0.1 M or NaOH 0.1 M.
Biosorption experiments
Metal ion binding experiments were performed by
incubation of 250 mg biomass (dry weight) with 50 mL
metals ions-containing solution in test tube on an orbital
rotary shaker at 120 rpm for a maximum of 120 minutes.
Samples were taken at given time intervals and then spin
dried at 5500 rpm for 15 minute. The experiment was
conducted at different pH values and were established by
adjusting it with HCl 0.1 M or NaOH 0.1 M solutions.
REV.CHIM.(Bucharest)♦ 67♦No. 7 ♦2016
 The data obtained in batch mode studies was used to
calculate the equilibrium metal adsorptive quantity. It was
calculated for each sample of Cu(II) and Pb(II) by using
the following expression:
qe = v (c0 – ce) / m (1)
where qe is the amount of metal ions adsorbed onto per
unit weight of beer yeast in (µg g-1), v is the volume of the
solution in (L) and m is the weight of the dry biomass used
in (g), c0 and ce are the initial and metal equilibrium
concentration in (µg L-1).
Effect of contact time on biosorption
Batch biosorption tests were done at different contact
time at the initial concentration of 963 µg L-1 for Cu(II) and
986 µg L-1 for Pb(II) respectively and brewer yeast dose
concentration 5 g L-1 in 50 mL solution. The pH established
was 6. Test tubes were agitated on an orbital rotary shaker
at 120 rpm, then the samples were collected at different
intervals (5, 15, 30, 60, 90 and 120 min.) and were
centrifuged. The concentrations of metal ions in the
supernatant solution were analyzed using UV-Vis diode
array spectrometer (Hewlet-Packard). Each determination
was repeated three times and the given results were the
average values. The standard deviation is less than 5%.
Effect of pH solution on biosorption
For metal ions biosorption the pH is one of the most
important environmental factor. The effect of pH on the
adsorption capacity of brewer yeast was investigated using
solutions of 963 µg L-1 for Cu(II) and 986 µg L-1 for Pb(II),
and brewer yeast dose concentration 5 g L-1 in 50 mL
solution. The pH values ranged from 3.5 to 8 at 293 K.
Experiments could not be performed at higher pH value
due to the possibility of the lead ions precipitation. Test
tubes were agitated on an orbital rotary shaker at 120 rpm
for 60 min to ensure that the equilibrium was reached.
Then the mixtures were centrifuged and the concentration
of metals in the supernatant was measured.
Effect of yeast dose on biosorption
Batch sorption test were done at different yeast
concentration between 0.1 g/100 mL and 1.5 g/100 mL at
pH=6, for a contact time of 60 min. The initial concentration
of Cu(II) and Pb(II) were 963 µg L-1 for Cu(II) and 986 µg
L -1 for Pb(II). The mixtures were centrifuged and the
concentration of metals in the supernatant was measured.
Sorption dynamic studies
Samples of biomass were placed into a series of 250
mL conical flasks followed by addition of 150 mL copper
and lead solution concentration of 1, 25, 50, 75 and 100
mg L-1. The brewer yeast concentration was established at
2 g L-1. The mixture were adjusted at pH=6 using 0.1 M
NaOH and 0.1 M HCl solutions. The sorption equilibrium
data were analyzed using the Langmuir adsorption
isotherms. Langmuir adsorption isotherm is often used to
describe maximum adsorption capacity of an adsorbent and
it is given as:
qe = qm x KL x Ce / 1 + KL x Ce (2)
where qe (mg g-1) is the adsorption amount of adsorbent at
equilibrium; qm (mg g-1) is the maximum adsorption
amount of metal ions, Ce (mg L-1) is the equilibrium
concentration of adsorbate in solution and KL (L mg-1) is
the equilibrium adsorption constant which is related to the
affinity of the binding sites. The Langmuir constants KL and
qm are calculated with the following equation:
REV.CHIM.(Bucharest)♦67 ♦ No. 7 ♦ 2016 http://www.revistadechimie.ro
Ce/qe = 1 / KL x qm + qe/qm (3)
where Ce (mg L-1) is the equilibrium concentration of
adsorbate in solution; qe (mg g-1) is the adsorption amount
of adsorbent at equilibrium, qm (mg g-1) is the maximum
adsorption amount of metal ions and KL (L mg-1) is the
equilibrium adsorption constant which is related to the
affinity of the binding sites.
Metal ions quantification by UV-Vis spectroscopy
After the biosorption experiments were completed,
metal concentrations were determinated using UV-Vis
spectroscopy. UV-Vis spectra were recorded on an UV-Vis
diode array spectrometer (Hewlet-Packard). For copper
and lead quantification it was used a method according to
Szabo et al. (2011), which consist in the complexation of
these metal ions with 4-(2-pyridylazo) resorcinol (PAR)
[16]. This ligand is a non-selective azo dye, widely used as
colorimetric reagent for metal ions because of forming
very stable, water-soluble and highly colored complexes
with the majority of transition metals [17]. For the metal
complex preparation, copper sulphate and lead nitrate of
10-4 M concentrations were mixed with 10-4 M PAR solution
at 1:2 molar ration resulting in Cu(PAR)2 and Pb(PAR)2
complexes. According to literature data, PAR forming
bidentate complexes with copper and lead at pH above 5
[17]. The atoms involved in the coordination are pyridine
nitrogen, the azo nitrogen farthest from the heterocyclic
ring and the o-hydroxyl group [17]. The UV-Vis spectroscopy
is one of the most used method for the detection and
quantification of metallic ions. The UV-Vis spectra of
copper and lead ions with PAR show a broad band for
metal-ionofor complexes (fig. 1). The PAR molecule
present the absorption maximum at 412 nm, while for
Cu(II) complex this maximum is located at 505 nm and
for Pb(II) at 515 nm. For quantitative determination of Cu(II)
and Pb(II) ions in the supernatant after biosorption it was
used calibration curves based on the absorbance of these
complexes at their maxima (fig. 1).
Fig. 1. The UV-Vis spectra of PAR, Pb(PAR)2 and Cu-(PAR)2
complexes
In figure 2 it is presented the calibration curve for Cu-
(PAR) 2 complex. The squares (R 2) of the correlation
coefficient of the calibration curves were found to be 0.9808
for copper complex and 0.9796 for lead complex.
Results and dicussions
The effect of contact time
The effect of contact time on copper and lead ions
biosorption by brewer’s yeast was studied and the results
are presented in the figure 3. It was found that the
adsorptive capacity qe of both copper and lead ions on
1277
 Fig. 2. Calibration curve for Cu-(PAR)2 complex
Fig. 3. The effect on contact time on biosorption
brewer yeast increase with the increasing of the contact
time until the equilibrium stage was obtained (fig. 3).
The biosorption process was rapid in the first 15 min.
and the equilibrium was reached after 60 min. with a very
slow increase until 90 min. This trend emphasizes that
sorption time play an important role on recovery efficiency,
which decreases with the increasing of contact time with
metal solution. Metal accumulation inside the cells may
resulted from bioaccumulation, slow dependent removal
mechanism, or by simple metal diffusion [19, 20].
Han et al. (2006) reported that the uptake of metal ions
by microorganisms in batch system has been shown to
occur in two stages: an initial rapid stage (passive stage),
followed by a slower process (active stage) [15]. The first
stage represent a physical adsorption or ion-exchange at
the surface of the biomass, which is biosorption. The
biosorption equilibrium occurs at the end of the physical
adsorption stage.
The effect of pH
The charge of the adsorbent and the adsorbate depends
on the pH of the solution, thus the adsorption of copper and
lead ions as a function of pH was measured, and the results
are presented in the figure 4. The pH values chosen for
experiments were 3.5, 5, 6, 7 and 8. There was an increase
in biosorption capacity of biomass with the increasing of
the pH until pH=6 for both metals. The highest metal
uptake values obtained for copper and lead ions were 69.1
µg g-1 and 78.8 µg g-1 respectively. The low level of Pb(II)
and Cu(II) uptake at lower pH values could be attributed to
the increased concentration of hydrogen and hydronium
ions competing for metal ions binding sites on the biomass.
Fig. 4. The pH effect on Cu(II) and Pb(II) ions biosorption by
brewer’s yeast
1278 http://www.revistadechimie.ro
Fig. 5. Effect of yeast dosage on biosorption of Cu(II) and Pb(II)
ions
The increase in the Pb(II) and Cu(II) biosorption capacity
at the higher pH values (between 5 and 7) may be explained
by the ionization of functional groups on the cell surface
which serve as binding sites related to the isoelectric point
of the cells. The decreasing in the biosorption capacity at
higher p H (>7) is due to the formation of soluble
hydroxylated complexes of the metal ions, especially for
Pb(II) ions, and their competition with the active sites, and
as a consequence, the retention would decrease again. At
pH values above the isoelectric point, there is a net negative
charge on the cell wall components and the ionic state of
ligands such as carboxyl, amino, phosphate groups could
promote reactions with metal cations.
Effect of yeast dosage
The effect of yeast concentration on removal efficiency
is presented in the figure 5. Metal removal percentage (R%)
was calculated by equation:
R (%) = (C0-Ct) x 100 / C0 (4)
where (R%) is the ratio of difference in metal
concentration before and after adsorption; Co is the initial
concentration of metal (µg L -1 ); C t is the residual
concentration of metal after adsorption had taken place
over a period of time t (µg L-1).
Results showed that the percent of metal removal
increase as yeast dosage increase up to 1 g/100 mL solution
for metal ions. The increase in metal uptake with yeast
dosage confirms the increase in binding sites for metal
ions. In both experiments it was observed that the metal
uptake decrease with the increasing of yeast dosage over
1 g/100 mL due to aggregation of the yeast [21]. The higher
uptake at lower biomass concentration could be due to
metal ions and biosorbent ratio, which decrease upon an
increase in biomass dosage. This fact could be attributed
to the formation of aggregates of biomass and may cause
interferences between binding sites at higher
concentration or insufficiently of metal ions in the solution
with respect to available binding sites. It is likely that protons
will then combine with metal ions for the ligands and
thereby decrease the interaction of metal ions with the
cell components.
Adsorption equilibrium
Adsorption isotherms show the distribution of solute
between the liquid and solid phases equilibrium conditions.
Langmuir model is probably the most popular isotherm
model due to its simplicity and its good agreement with
experimental data. Langmuir adsorption isotherm model
represents one of the first theoretical treatments of non-linear
sorption and suggests that the uptake occurs on a
homogenous surface by monolayer sorption without
interaction between adsorbed molecules. Although the
REV.CHIM.(Bucharest)♦ 67♦No. 7 ♦2016

Fig. 6. Langmuir adsorption model of Pb(II) biosorption on brewer
yeast biomass
Fig. 7. Langmuir adsorption model of Cu(II) biosorption on brewer
yeast biomass
Langmuir constant qmax is dependent on experimental
conditions such as pH solution and temperature, it is a
good measure for comparing different sorbents for the
same metal sorbate. The mono-component Langmuir
constant q max, represents the monolayer saturation at
equilibrium or the total capacity of beer yeast for copper
and lead ions.
The adsorption isotherms of Cu(II) and Pb(II) biosorption
on brewer yeast at pH=6 were shown in the figures 6 and
7, and the isotherms parameters are presented in table 1.
The Langmuir isotherm in terms of a dimensionless
constant separation factor RL is defined by:
R L = 1 / 1 + K L C0 (5)
where KL is the Langmuir constant and C0 is the initial metal
ions concentration .
The RL values between 0 and 1 indicate favorable
adsorption. The shape of isotherm is given by the value of RL
as given in table 2.
The data obtained represent a favorable adsorption in
the case of adsorption of Cu (II) and Pb(II) ions. KL (L
mg-1) and qmax (mg g-1) were determinated from the slope
and intercept of the Langmuir isotherm plots (fig. 6,7).
The maximum loading capacities of beer yeast was
obtained as 20.6 mg g-1 for Cu(II) ions and 22.9 mg g-1 for
Pb(II) ions, so the ability of lead adsorption is higher than
copper ions (table 1), which is in accordance with Han et.
al [15].
Table 1
LANGMUIR ISOTHERMS PARAMETERS FOR Cu(II) AND Pb(II)
BIOSORPTION ON BREWER YEAST BIOMASS
REV.CHIM.(Bucharest)♦67 ♦ No. 7 ♦ 2016 http://www.revistadechimie.ro
Table 2
CORRESPONDENCE
BETWEEN SEPARATION
FACTOR RL VALUES AND
LANGMUIR ISOTHERM
TYPE
Lower binding capacity of copper ions over lead ions on
the biomass could be explained by comparing the covalent
index of Pb(II) ion (11.1) with Cu(II) ion (5.67). The high
value of the covalent index shows the high degree of
binding capacity of the metal ions to the biological ligands.
Also, Pb(II) ion is classified as a class b ion, while Cu(II) is
classified as borderline ion, so the behavior of adsorption is
not full the same and the binding capacity is different.
Conclusions
The results of our experiments concluded that the low-
cost biomaterial, such brewer’s yeast, has the ability to
adsorb Cu(II) and Pb(II) ions from aqueous solutions.
The process of biosorption has reached equilibrium after
60 minutes and the optimum pH for both metals is 6. Results
showed that the percent of metal removal increase as
yeast dosage increase up to 1 g/100 mL solution for both
metal ions. Based on the correlation coefficients, it could
be concluded that Langmuir isotherm is suitable to describe
equilibrium data of Cu(II) and Pb(II) ions biosorption on
brewer yeast. The maximum loading capacities of beer
yeast was obtained as 20.6 mg g-1 for Cu(II) ions and 22.9
mg g-1 for Pb(II) ions.
Acknowledgements: This work was supported by a grant of the
Romanian Ministry of Education, CNCSIS – UEFISCDI, PN-II-RU-PD-
2012-3-0055, no. 47/30.04.2013.
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Manuscript received: 27.07.2015
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