Enzyme and Microbial Technology 41 (2007) 98–102
The investigation of lead removal by biosorption:
An application at storage battery industry wastewaters
Tolga Bahadir a,1 , Gulfem Bakan a,∗ , Levent Altas b,2 , Hanife Buyukgungor a,1
a
Environmental Engineering Department, Faculty of Engineering, Ondokuz Mayis University, Samsun, Turkey
b Environmental Engineering Department, Faculty of Engineering, Aksaray University, Aksaray, Turkey
Received 12 September 2006; received in revised form 1 December 2006; accepted 12 December 2006
Abstract
Lead is present in different types of industrial effluents, being responsible for environmental pollution. Biosorption has attracted the attention in
recent years as an alternative to conventional methods for heavy metal removal from water and wastewater. The biosorption of Pb(II) ions present
in the storage battery industry wastewaters intensively, by Rhizopus arrhizus has been investigated in this study. This microorganism has been
preferred since its biosorption feature was well known. A detailed study was conducted for the removal of Pb(II) ions which was very toxic even in
low quantities to the receiving environment, from storage battery industry wastewater by biosorption system as advanced treatment technique, and
to investigate the effects of the several parameters on its removal. The average Pb(II) ions concentration in the storage battery industry wastewater
found 3.0 mg/L and reducing this value below 0.5 mg/L was aimed. In this study, the effects of the media conditions (pH, temperature, biomass
concentration) on the biosorption of Pb(II) ions to R. arrhizus have been investigated in a batch reactor. Optimum biosorption conditions have
been found of initial pH 4.5, temperature 30 ◦ C and biomass concentration 1.0 g/L. The maximum biosorption capacity was obtained as 2.643 mg
Pb(II)/g microorganism.
© 2007 Elsevier Inc. All rights reserved.
Keywords: Biosorption; Storage battery industry wastewater; Lead removal; Rhizopus arrhizus
1. Introduction
Heavy metal pollution is an environmental problem of worldwide concern. Heavy metals released into the environment have
been increasing continuously as a result of industrial activities
and technological development, posing a significant threat to the
environment and public health because of their toxicity, accumulation in the food chain and persistence in nature. The heavy
metals lead, mercury, copper, cadmium, zinc, nickel, chromium
are among the most common pollutants found in industrial effluents. Even at low concentrations, these metals can be toxic to
organisms, including humans. For instance, lead is extremely
toxic and can damage to the nervous system, kidneys and reproductive system, particularly in children [1,2].
∗
Corresponding author. Tel.: +90 3623121919; fax: +90 3624576035.
E-mail addresses: [email protected] (T. Bahadir), [email protected]
(G. Bakan), lvnt [email protected] (L. Altas),
[email protected] (H. Buyukgungor).
1 Tel.: +90 3623121919; fax: +90 3624576035.
2 Tel.: +90 3822150953; fax: +90 3822150592.
0141-0229/$ – see front matter © 2007 Elsevier Inc. All rights reserved.
doi:10.1016/j.enzmictec.2006.12.007
Heavy metals are discharged from various industries, such
as smelters, electroplating, metal refineries, textile, mining,
ceramic and glass. Many industries, such as coating, automotive, storage batteries, aeronautical and steel industries generate
large quantities of wastewater containing various concentrations
of lead [2–5].
Lead is particularly suitable for batteries, because of its specific characteristics (conductivity, resistance to corrosion and
the special reversible reaction between lead oxide and sulphuric
acid). A recent survey of three storage battery producers showed
that the pH of wastewater at the source ranged between 1.6 and
2.9, while the concentration of soluble lead was in the range
of 5–15 mg/L. The storage battery industry wastewater contains
lead(II) ion, is occurred from formation, negative and positive
drying operations [6].
Conventional methods used to remove dissolved heavy
metal ions from wastewaters include chemical precipitation,
chemical oxidation and reduction, ion exchange, filtration,
electrochemical treatment and evaporative recovery. However,
these high-technology processes have significant disadvantages,
including incomplete metal removal, requirements, such as
T. Bahadir et al. / Enzyme and Microbial Technology 41 (2007) 98–102
expensive equipment and monitoring systems, high reagent,
energy and generation of toxic sludge or other waste products
that require disposal. New technologies are required that can
reduce heavy metal concentrations to environmentally acceptable levels at affordable costs [2,7,8].
Biosorption process offers the advantages of low operating
costs, possibility of metal recovery, regeneration of biosorbent,
minimization of the volume of chemical and/or biological sludge
to be disposed of and high efficiency in detoxifying very dilute
effluents [9,10]. Biosorption of heavy metals is one of the most
promising technologies involved in the removal of toxic metals
from industrial waste streams [11].
Biosorption could be generally defined as the removal of
metal or metalloid species, compounds and particulate from
solution by biological materials. The term “biosorption” is inadequate since it encompasses, metal adsorption and chemical
deposition as well as intracellular uptake by cells. Microorganisms including bacteria, algae, fungi and yeast can efficiently
accumulate heavy metals and radio nucleides from their external
environment [12].
Rhizopus arrhizus, a filamentous fungus, is used in fermentation industries to produce various metabolites, such as lipase,
fumaric acid, lactic acid, steroids and celluloses [13]. The inactive biomass of R. arrhizus is effective in the removal of heavy
metals, separately radioactive elements, such as uranium and
thorium [14]. The cell wall of R. arrhizus essentially consists
of various organic compounds including chitin, acidic polysaccharides, lipids, amino acids and other cellular components
[15].
The main aim of this research was to determine the effects
of different parameters (i.e. pH, temperature, biomass concentration) which were effective at the removal of Pb(II) ions from
storage battery industrial wastewaters using non-living biomass
of R. arrhizus by biosorption.
2. Materials and methods
2.1. Microorganism and growth conditions
99
Table 1
Characteristics of wastewater used
Characteristics
Value
pH
Lead (mg/L)
Nickel (mg/L)
Copper (mg/L)
Chromium (mg/L)
Zinc (mg/L)
Cobalt (mg/L)
Cadmium (mg/L)
1.53–1.62
3.094
0.005
0.027
0.373
–
–
–
(−) Not available.
measured as 42 ◦ C. The daily flow of the wastewater of the factory was changing in between 96 and 264 m3 /day according to the production variations. The
concentrations of the other metal ions present at the wastewater (Table 1) were
so small that they could not effect the removal of lead ions from the wastewater.
In fact, optimum removal conditions of metal ions were different. In this study,
the main aim was to remove the lead(II) ions from the storage battery industry
wastewaters.
2.4. Biosorption studies
The microorganism solutions (5 mL) were added to Erlenmayer flasks containing wastewater of storage battery industry (45 mL). The flasks were agitated
at 120 rpm on a shaker (Nüve ST 402) for 90 min, which gave ample time for
adsorption equilibrium to be reached at the desired conditions. After the samples
were filtered by Whatman (pore size 45 ␮m) the concentration of unadsorbed
lead(II) ion in the supernatants were measured by using an atomic adsorption
spectrophotometer (AAS: ATI-UNICAM 929).
Metal uptake (q) was determined as follows:
q=
V (Ci − Ceq )
m
(1)
where q (metal uptake, mg/g) is the amount of metal ions adsorbed on the
biosorbent, V (mL) the volume of metal containing solution in contact with
the biosorbent, Ci (mg/L) the initial concentration of metal ions in the solution, Ceq (mg/L) the final concentration of metal ions in the solution and m
(g) is the dry weight of fungal biomass. These are schematically shown in
Fig. 1.
R. arrhizus, a filamentous fungus, was obtained from the US Department
of Agriculture Culture Collection (NRRL 2286). R. arrhizus was grown at
room temperature in liquid media containing malt extract (Merck) (17.0 g/L)
and soybean peptone (Acumedia) (5.4 g/L).
2.2. Preparation of the microorganism for biosorption
After the growth period (5–6 days), R. arrhizus was washed twice with
distilled water, inactivated using 1% formaldehyde (Merck) and then dried in an
oven at 70 ◦ C for 24 h. Dried R. arrhizus was homogenized in a mixer (Severin)
to destroy cell aggregates and sieved at Fritsch (Alnalysette 3, model SPARTAN)
sieve. Different particle ranges of biomass, such as 0.045–0.063, 0.063–0.090
and 0.090–0.125 mm were used for this metal biosorption study. Finally, for
adsorption studies, a weighed amount of dried cells (10 g/L) were suspended in
100 mL of distilled water.
2.3. Wastewater sample
Wastewater was obtained from storage battery factory in Industrial Park
Region Administrations, Yiğit Battery Co., Inc., Ankara, Turkey. The composition of the wastewater is given in Table 1. The temperature of the wastewater was
Fig. 1. Experimental set up for the removal of Pb(II) ions from aqueous solutions.
100
T. Bahadir et al. / Enzyme and Microbial Technology 41 (2007) 98–102
Fig. 2. Effect of initial pH on the equilibrium Pb(II) uptake in the storage battery industry (temperature, 30 ◦ C; biomass concentration, 1.0 g/L; agitation rate,
120 rpm; contact time, 90 min).
3. Results and discussion
Fig. 4. Effect of biomass concentration on the equilibrium Pb(II) uptake in
the storage battery industry (initial pH, 4.5; temperature, 30 ◦ C; agitation rate,
120 rpm; contact time, 90 min).
3.1. Effect of initial pH on lead(II) removal
3.3. Effect of biomass concentration on Pb(II) removal
The most important single parameter influencing the sorption capacity is the pH of adsorption medium. The initial pH
of adsorption medium is related to the adsorption mechanisms
onto the adsorbent surface from water and reflects the nature of
the physicochemical interaction of the species in solution and
the adsorptive sites of adsorbent [7]. Pb(II) uptake from storage battery industry wastewater was very sensitive to changes
in biosorption medium pH. To determine the effect of pH of
the biosorption medium on the initial biosorption rates of Pb(II)
ions, the pH was varied between 2.0 and 5.0. The optimum initial pH of the biosorption medium was found to be 4.5 (Fig. 2).
The equilibrium Pb(II) uptake was 2.265 mg/g at this pH value.
At pH higher than 5.0, lead(II) ions precipitated and adsorption
studies at these pH values could not be performed.
In order to determine the effect of biomass concentration on
Pb(II) removal, stock solutions of 10.0 g/L were prepared for different microorganism particle size ranges, such as 0.045–0.063,
0.063–0.090 and 0.090–0.125 mm. These stock solutions were
diluted to 0.1–1.0 g/L standard solutions in order to find the
effect of microorganism concentration on biosorption (Fig. 4).
For Pb(II) adsorption on R. arrhizus, as the concentration
of microorganism was increasing at the medium, the Pb(II) ion
concentration was decreasing (Fig. 4). At equilibrium conditions, biosorption amount of Pb(II) was increasing parallel to
microorganism concentration. When the Pb(II) ions remained
on the solution concentration was investigated at each particle
size solutions, the optimum biomass concentration was obtained
at 1.0 g/L dose. It may be possible to consider the biomass dose as
0.5 g/L but it may take time to reach at equilibrium conditions at
this dose. After sieving, microorganisms at 0.063–0.090 mm particle size, mainly at medium size were selected for the removal
of lead ions storage battery industrial wastewater.
3.2. Effect of temperature on lead(II) removal
Results of removal of Pb(II) ions from storage battery industry wastewater experiments carried out at different temperatures
ranging from 20 to 45 ◦ C are shown in Fig. 3. At low temperatures, the binding of Pb(II) ions to R. arrhizus was by passive
uptake. Maximum initial adsorption rate of Pb(II) ions to R.
arrhizus was obtained at temperature 30 ◦ C. The equilibrium
Pb(II) uptake was 2.306 mg/g at this temperature value.
Fig. 3. Effect of temperature on the equilibrium Pb(II) uptake in the storage
battery industry (initial pH, 4.5; biomass concentration, 1.0 g/L; agitation rate,
120 rpm; contact time, 90 min).
3.4. Wastewater treatment of storage battery industry
For the treatment of storage battery industry wastewater, different samples of wastewater were contacted with the biomass
for 90 min at optimum conditions. The variation of Pb(II) ion
concentration at the solutions remained and the amount of pollutant adsorbed per unit weight of microorganisms through the
time is illustrated in Fig. 5.
In the biosorption of the tested metal, Pb(II) by the different R. arrhizus dose, most of the metal ions were sequestered
very fast from solutions within the first phase (5 min) and the
concentration of Pb(II) ions at the wastewater was 0.369 mg/L
and removal efficiency was performed as 86.82% at this stage
(Fig. 5). After this time interval, the biosorption process was
nearly very slow at the second phase. It can be concluded from
Fig. 5 that the biosorption process has reached at equilibrium at
time 60 min and the biosorption of the Pb(II) ions from storage
battery industry wastewater was 94.39% at this time intrude.
As a conclusion, it is therefore important to study new methods for metal exclusion and/or recuperation from dilute solutions
T. Bahadir et al. / Enzyme and Microbial Technology 41 (2007) 98–102
101
Table 2
Comparison of various literature studies of metal removal by biosorption
Biosorbent
Metal
Operation conditions
Co
pH
T
(◦ C)
Biomas
concentration (g/L)
Agitation rate
(rpm)
Biosorption
capacity q
Reference
Sargassum sp.
Pb
Ni
1.0 mmol/L
1.0 mmol/L
5.0
5.5
25
25
1
1
200
200
1.16 mmol/g
0.61 mmol/g
[1]
Panida sp.
Pb
Ni
1.0 mmol/L
1.0 mmol/L
5.0
5.5
25
25
1
1
200
200
1.25 mmol/g
0.63 mmol/g
[1]
Phanerochaete chrysosporium
Cu
Zn
Pb
100 mg/L
100 mg/L
100 mg/L
6.0
6.0
6.0
25
25
25
1
1
1
100
100
100
60.94 mg/g
34.13 mg/g
76.95 mg/g
[3]
Activated sludge
Saccharomyces cerevisiae
Cr
Cd
Pb
100 mg/L
25 mg/L
25 mg/L
1.0
6.0
5.0
25
30
30
0.5
1
1
150
200
200
61.00 mg/g
15.38 mg/g
15.59 mg/g
[7]
[10]
Rhizopus arrhizus
Cd
Pb
Cu
3.5
4.0
4.5
30
25
30
n.a.
n.a.
n.a.
150
150
150
11.17 mg/g
12.31 mg/g
7.32 mg/g
[15]
Chlorella vulgaris
Cu
Ni
Cr
100 mg/L
100 mg/L
100 mg/L
4.5
5.0
2.0
25
25
25
1
1
1
125
125
125
40.00 mg/g
42.30 mg/g
23.00 mg/g
[16]
Scenedesmus obliquus
Cu
Ni
Cr
100 mg/L
100 mg/L
100 mg/L
4.5
5.0
2.0
25
25
25
1
1
1
125
125
125
20.00 mg/g
18.70 mg/g
15.60 mg/g
[16]
Synechocystis sp.
Cu
Ni
Cr
100 mg/L
100 mg/L
100 mg/L
4.5
5.0
2.0
25
25
25
1
1
1
125
125
125
23.40 mg/g
15.80 mg/g
19.20 mg/g
[16]
Bacillus sp.
Pb
Cu
250 mg/L
200 mg/L
3.0
5.0
25
25
2
2
125
125
92.27 mg/g
16.25 mg/g
[17]
Rhizopus arrhizus
Pb
4.5
30
1
120
2.64 mg/g
50 mg/dm3
50 mg/dm3
50 mg/dm3
2.80 mg/L
This work
n.a., not available.
(1–100 mg/L) and for the reduction of heavy metal ions very
low concentrations [2]. The removal of Pb(II) ions obtained
from storage battery industry wastewater was performed by R.
arrhizus fastly and efficiently. Biosorption results of heavy met-
als reported by the other researchers in the literature by various
biosorbents at various operation conditions are summarized in
Table 2. It can be stated that the uptake values determined in this
study were found to be higher than that of many similar biosorbents. So, it can be pointed out that R. arrhizus is a good and
effective biosorbent that can be used for the removal of Pb(II)
ions from storage battery industrial wastewater at the present
conditions of this study.
The discharge limit of wastewater for storage battery industry
wastewater is given as 2.00 mg/L at National Environmental Regulations of Turkey. At this study, after the treatment
of wastewater, the Pb(II) concentration was measured as
0.157 mg/L which was far from the maximum discharge limit
of legacy. According to this results, it can be stated as that
the storage battery industry wastewaters at low lead concentration can effectively be treated by biosorption of Pb(II) using
R. arrhizus.
Acknowledgements
Fig. 5. Variations of Pb removal by biosorption of R. arrhizus from storage
battery industry wastewaters biosorption as a function of time (initial pH,
4.5; temperature, 30 ◦ C; biomass size, 0.063–0.090 mm; biomass concentration,
1.0 g/L; agitation rate, 120 rpm).
We would like to thank to Sadık Çelikel and Yiğit Battery
Co., Inc., for the storage battery industry wastewater samples,
and Zümriye Aksu and Tülin Kutsal for the R. arrhizus microorganism.
102
T. Bahadir et al. / Enzyme and Microbial Technology 41 (2007) 98–102
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