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Waste Management 28 (2008) 333–338

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Bioleaching of metals from spent lithium ion secondary batteries

using Acidithiobacillus ferrooxidans
a,b,*
Debaraj Mishra , Dong-Jin Kim a, D.E. Ralph c, Jong-Gwan Ahn a, Young-Ha Rhee b

a
Korea Institute of Geosciences and Mineral Resources, Gajeong-dong, Daejeon 305-350, South Korea
b
Department of Microbiology, Chungnam National University, Kung-dong, Daejeon 305-764, South Korea
c
A J Parker CRC for Hydrometallurgy, Chemistry Department, Murdoch University, South Street, Murdoch, 6150, Western Australia, Australia

Accepted 19 January 2007

Available online 21 March 2007

Abstract

Bioleaching of spent lithium ion secondary batteries, containing LiCoO2, was attempted in this investigation. The present study was
carried out using chemolithotrophic and acidophilic bacteria Acidithiobacillus ferrooxidans, which utilized elemental sulfur and ferrous
ion as the energy source to produce metabolites like sulfuric acids and ferric ion in the leaching medium. These metabolites helped dis-
solve metals from spent batteries. Bio-dissolution of cobalt was found to be faster than lithium. The effect of initial Fe(II) concentration,
initial pH and solid/liquid (w/v) ratio during bioleaching of spent battery wastes were studied in detail. Higher Fe(II) concentration
showed a decrease in dissolution due co-precipitation of Fe(III) with the metals in the residues. The higher solid/liquid ratio (w/v) also
affected the metal dissolution by arresting the cell growth due to increased metal concentration in the waste sample. An EDXA mapping
was carried out to compare the solubility of both cobalt and lithium, and the slow dissolution rate was clearly found from the figures.
 2007 Elsevier Ltd. All rights reserved.

1. Introduction uti et al., 1998). In these processes, microorganisms (bacte-

Production of spent lithium ion secondary batteries
(LIBs) is increasing world-wide and 10% of market share
is in Korea. The spent lithium ion batteries contain 5–
15 wt.% cobalt and 2–7 wt.% lithium as important constit-
uents, which are an active cathodic material (Lee and Rhee,
2002). Since cobalt is a relatively expensive material com-
pared to the other battery constituents, its recovery is one
of the primary objectives in the recycling of spent batteries.
Lithium has also vital importance in many industrial appli-
cations. To recover cobalt and lithium from battery wastes,
hydrometallurgical operations have been adopted (Nan
et al., 2005; Zhang et al., 1998; Shin et al., 2005). However,
it has been reported that bio-hydrometallurgical processes
have been gradually replacing the hydrometallurgical one
due to their higher efficiency, lower costs and few industrial
requirements (Ehrlich, 1981; Rohwerder et al., 2003; Cerr-
*
Corresponding author. Tel.: +82 42 868 3592; fax: +82 42 861 9727.
E-mail address: mishradebaraj@yahoo.com (D. Mishra).
ria and fungi) perform numerous physiologically important
reactions that enable them to grow and reproduce. The for-
mations of metabolic products such as inorganic and
organic acids by microbial activities help in the leaching
of metals from wastes and minerals. Bio-hydrometallurgi-
cal processing of solid waste is similar to natural biogeo-
chemical metal cycles and reduces the demand of
resources, such as ores, energy and landfill space (Verstra-
ete, 2002; Rossi, 1990; Horvath and Gruiz, 1996; Moore
and Luoma, 1990; Stefan and Nicole, 2002).
A key microorganism is the iron and sulfur oxidizing
bacterium, Acidithiobacillus ferrooxidans, which has been
studied extensively for its ability to oxidize metal sulfide
ores (Norris, 1990). However, its application in the dissolu-
tion of metals from various industrial waste products is a
core topic at present. Use of this bacterium has been cited
in the dissolution of valuable metals from wastes such as
electronics scrap, spent nickel–cadmium batteries, smelter
dusts and ferromanganese crusts (Brandl et al., 2001; Zhu
et al., 2003; Olson et al., 1990; Nakazawa and Sato,

0956-053X/$ - see front matter  2007 Elsevier Ltd. All rights reserved.
doi:10.1016/j.wasman.2007.01.010
334 D. Mishra et al. / Waste Management 28 (2008) 333–338
1995). Taking this into account, it was thought that it
should be possible to use a bioleaching method for the
extraction of cobalt and lithium from spent lithium ion sec-
ondary batteries.
2. Materials and methods
All the reagents used in the experiment are of analytical
grade.
2.1. Bacteria
A pure culture of A. ferrooxidans (ATCC 19859) was
obtained from EWHA University, Seoul. The organism
was cultured at initial pH 2.0 in iron free 9 K medium (Silv-
erman and Lundgren, 1959) with elemental sulfur as energy
source. Cells were harvested at late logarithmic phase when
the culture pH was about 1.2 and re-suspended with iron-
free 9 K medium and used as inocula in the different leach-
ing experiments.
2.2. Adaptation of bacteria
The growth cultures of A. ferrooxidans on sulfur was
investigated with the addition of LIBs at 30 C and
200 rpm having the standard growth medium (iron free)
(Silverman and Lundgren, 1959). Initial growth was carried
out at a LIB concentration of 0.25 g/100 mL. The bacterial
growth was monitored by cell counts (Neuber counting
chamber) and pH measurement. The cell number was con-
tinuously monitored during the growth period. The num-
ber remained almost constant after 2 weeks and then the
further addition of LIBs in the same medium was followed.
The amount of LIBs was slowly increased in the medium to
1 g/100 mL. Since the waste material is an oxide in nature,
the cell number was observed to decrease slowly at 1 g/
100 mL concentration of the LIBs. Expecting the harmful
effects of cobalt and lithium on this pure bacterium, we lim-
ited our adaptation study at 10 g/L. However, we did not
study the uptake capacity of cobalt and lithium of this pure
culture of A. ferrooxidans. At the final growth stage, the
suspended bacterial population was found to be at least
4.0 · 107 cells/mL. This adapted strain was used in our
bioleaching experiments.
2.3. Characterization of battery waste powder
Generally lithium ion batteries (LIBs) consist of some
heavy metals like cobalt, nickel, lithium and some other
organic chemicals, and the compositions also vary slightly
with different manufacturers. In the present study, cathode
waste materials of LIBs, previously separated by the man-
ufacturers, were collected. The material was ground by
milling and sieved to obtain a mesh size of <150 lm. This
powder was characterized by XRD (Rigaku X-Ray Dif-
fractometer, Rigaku, Japan) and the LiCoO2 phase was
clearly viewed. The specific surface area of the waste sam-
ple was determined by BET method (Micromeritics, Tristar
3000, Unit 1) and found to be 32.6 m2/g. The mean particle
size was measured with a particle size analyzer (Malvern
Mastersizer 2000, Version 2.00) and found to be
d(0.5) = 8.3 lm.
2.4. Bioleaching experiment
Bioleaching studies were carried out in 300-mL conical
flasks. About 90 mL of 9 K medium was prepared and
the initial pH was adjusted to 2.5 using concentrated
H2SO4. To the prepared medium, 1.0 g of powdered ele-
mental sulfur and Fe(II) ion of 3 g/L were input as the sole
energy source for the bacterium. For the leaching experi-
ments, the required amount of LIBs was added, followed
by 10 mL of inoculum, having a bacteria concentration
of 3.2 · 107 cells/mL. The flasks were incubated in a shak-
ing incubator at 30 C and agitated at 200 rpm. Sterile con-
trols were performed using filter-sterilized medium at an
initial pH of 2.5 with 1% of elemental sulfur and 3 g/L of
Fe(II) ion in absence of inoculum.
2.5. Analytical determinations
The pH of each experiment was measured periodically
throughout the experimental period using a glass electrode
(Thermo Electro Corporation, Orion 720+). The leached
solutions (1.5 mL) were taken periodically, transferred to
microfuge tubes (1.5 mL) and centrifuged at 12,000 rpm
for 10 min. Cobalt and lithium concentrations in the solu-
tion were determined after centrifugation using an Atomic
Absorption Spectrophotometer (Varian SpectrAA-400).
Samples were diluted to the appropriate concentration
range with distilled water and measured in triplicate.
Losses from leaching solutions during leaching experiments
were compensated by adding distilled water.
3. Results and discussion
3.1. Metal solubilization by bacteria
An initial series of experiments was carried out to study
the behavior of the microbial strain in the LIB wastes con-
taining no energy source such as sulfide or sulfur or iron
for the microbial growth. The preliminary results are pre-
sented in Figs. 1 and 2, where data showing the percentage
of Co and Li leached into the solution are plotted, at solid/
liquid (w/v) ratio of 5 g/L. Each experiment showed a high
dissolution of cobalt in the leaching medium compared to
lithium. From the results, microbial activity is confirmed
during leaching experiments. Where inocula were present
with energy sources such as elemental sulfur and Fe(II)
ion, dissolution of both lithium and cobalt increased. In
the presence of bacteria, cobalt leaching was found to be
65%, whereas it was 41% in the absence of cells. Also,
microbial activity during the LIB dissolution experiments
was confirmed by comparing the pH change (Fig. 3) of
 D. Mishra et al. / Waste Management 28 (2008) 333–338 335

70 ure of the material as the materials have oxide

60
compositions.
% of Cobalt leached

During bacterial attack, the cells metabolize elemental

50 sulfur to H2SO4 and the protons from this metabolite
40
attack or react with the metals in LIBs. Being both an iron
and sulfur oxidizer, A. ferrooxidans forms sulfate from
30 reduced elemental sulfur in the leaching medium. So, the
20
generation of sulfuric acid and its attack on the metals in
LIBs occurred in parallel. That is why in the absence of sul-
10 fur and iron, the bacteria were inactive and effective cell
0
density was quite low in the lixiviant. The oxide compo-
Blank Inoculum(only) Iron+Sulfur Iron+Sulfur+Inoculum
nents of the metals in the LIBs are expected to suppress
Fig. 1. Cobalt leaching at different experimental conditions. (Conditions: the cell growth in absence of the vital energy sources, and
solid/liquid ratio (w/v) – 5 g/L, initial pH – 2.5, elemental sulfur – 1%, hence the pH of the solution increased to 5.5–6.0. Thus,
Fe(II) – 3 g/L). a low dissolution of cobalt and lithium was obtained at
the end.
10
3.2. Metal dissolution at different initial Fe(II)
8
concentrations
% of Li leached

All metal sulfides leaching by A. ferrooxidans involve

6
4
2
0
Blank
Fig. 2. Lithium leaching at different experimental conditions. (Conditions:
solid/liquid ratio (w/v) – 5 g/L, initial pH – 2.5, elemental sulfur – 1%,
Fe(II) – 3 g/L).
the leaching solutions and blank during the course of the
experiment. In the presence of bacteria, the pH of the solu-
tion decreased from 2.5 to 1.8 due to production of sulfuric
acid by the oxidative action of bacteria, whereas in blanks,
the pH of the solution increased from 2.5 to 4.5. The
increase in pH in case of blanks is due to the alkaline nat-
Inoculum (Only) Iron+ Sulfur
Fe(II) ion as a cardinal part in the leaching medium. Since
the present experiment used pure culture of A. ferrooxi-
dans, the effect of Fe(II) ion was considered having its role
during cobalt and lithium leaching from the LIBs. Addi-
tion of Fe(II) ion only acted as a source for bacterial
growth during leaching process. The oxidation of ferrous
Iron+ Sulfur to ferric could not produce any oxidative attack on the
+Inoculum
metal surface or in the other sense there was no indirect
leaching through Fe(III) ion in the solution. In the lixivi-
ant, cobalt and lithium are associated with their respective
sulfate components and here transformation of cobalt is
taking place from Co+3 ion to Co+2 ion. Therefore, the oxi-
dative attack of Fe(III) ion on reductive dissolution of
Co+3 is an unexpected critique. However, a partial solubi-
lization of lithium can be presumed via Fe(III) ion reaction
in the course of experiments. This can be surmised that
most of the dissolution of metals (Co and Li) was appar-
ently due to the formation of intermediate reducing com-
pounds, such as sulfates, by sulfur metabolism of

6
60 Fe(II)= 1 g/L
Fe(II)= 3 g/L
Fe(II)= 5 g/L
pH of the solution

5 50
% of Cobalt leached

Fe(II)= 7 g/L
Control
40
4
Blank
Inoculum(only) 30
Iron+Sulfur
3 Iron+Sulfur+Inoculum
20

2
10

0 2 4 6 8 10 12 14 16 18 20 22
0
Time in days 0 2 4 6 8 10 12 14 16 18 20 22
Time in days
Fig. 3. Final pH of the solution at different experimental conditions.
(Conditions: solid/liquid ratio (w/v) – 5 g/L, initial pH – 2.5, elemental Fig. 4. Cobalt leaching at different Fe(II) concentration. (Conditions:
sulfur – 1%, Fe(II) – 3 g/L). solid/liquid ratio (w/v) – 10 g/L, initial pH – 2.5, elemental sulfur – 1%).
336 D. Mishra et al. / Waste Management 28 (2008) 333–338

bacteria. Figs. 4 and 5 illustrate the pattern of metal (Co 60 pH 2.5

and Li) dissolution during the bioleaching of LIBs at differ- pH 3.0
pH 3.5
ent initial Fe(II) concentrations (1, 3, 5 and 7 g/L), while 50 pH 4.0

% of Cobalt leached
keeping the concentration of elemental sulfur constant at
40
1%. The Fe(III) ion in the leaching medium finally precip-
itated with the metals in the waste materials. It made the 30
cobalt dissolution slower forming complex components in
association with cobalt (from XRD data). During XRD 20

screening, even some phases of jarosites were obtained at

10
higher Fe(II) ion concentration and the presence of such
materials are detrimental for bacterial growth (Curutchet 0
0 5 10 15 20 25
et al., 1992). Therefore, assuming that lower Fe(II) ion con- Time in days
centration would proliferate bacterial growth as well as
increase the metal dissolution in the leaching solution, we Fig. 6. Cobalt leaching at different initial pH. (Conditions: solid/liquid (w/
have carried out other experiments at 3 g/L of Fe(II) ion v) ratio – 10 g/L, initial Fe(II) – 3 g/L, elemental sulfur – 1%).
concentration.
12
3.3. Effect of initial pH pH 2.5
pH 3.0
10 pH 3.5

% of Lithium leached
pH 4.0
The acidity or pH of the medium usually controls bacte-
8
rial growth, and bacterial catalytic action is significant only
up to pH 3.5 (Pesic et al., 1989). In the present study we 6
have conducted experiments varying from pH 2.5–4.0.
Figs. 6 and 7 show the effect of initial pH of the medium 4
on the dissolution of Co and Li by bacterial leaching. At
pH 3.5 and 4.0, initial lag periods were observed for about 2

1 week and later a fast dissolution of cobalt was obtained.

0
Since at higher pH, Fe(II) ion oxidation by bacterial catal- 0 5 10 15 20
ysis is generally inhibited, therefore, in the beginning of Time in days
leaching experiments (at pH 3.5 and 4.0) slow metal disso-
Fig. 7. Lithium leaching at different initial pH. (Conditions: solid/liquid
lution was achieved due to retarded cell growth. However, ratio (w/v) – 10 g/L, initial Fe(II) – 3 g/L, elemental sulfur – 1%).
all inoculated flasks showed a gradual decrease in pH after
1 week of the experimental period due to the oxidation of
elemental sulfur to inorganic sulfuric acid. The solid sulfur 3.4. Metal dissolution at different solid/liquid ratio
was found slowly losing its hydrophobicity due to bacterial
action and obtained in the solution being wetted and dis- Bacterial dissolution of metals from battery wastes at
persed. In the sterile control flasks the sulfur compounds different solid/liquid (w/v) ratios is shown in Figs. 8
were in its original form (leaching data not shown). The and 9. Increasing the value of the solid/liquid (w/v) ratio,
final pH in all inoculated flasks was between 2 and 2.5 the dissolution of metal was decreased owing to bacterial
and independent of the initial value. An initial pH of 2.5
is suitable for leaching experiments.
70
0.5g
11 60 0.5g (Control)
Fe(II)=1 g/L
10 Fe(II)=3 g/L 1g
% of Cobalt leached

Fe(II)=5 g/L 50 1g (Control)

% of Lithium leaching

9 Fe(II)=7 g/L 1.5g

8 Control
40 1.5g (Control)
7 2g

6 2g(Control)
30
5
4 20

3
10
2
1
0
0 2 4 6 8 10 12 14 16 18 20 22 0 5 10 15 20
Time in days Time in days

Fig. 5. Lithium leaching at different Fe(II) concentration. (Conditions: Fig. 8. Cobalt leaching at different solid/liquid ratio (w/v). (Conditions:
solid/liquid ratio (w/v) – 10 g/L, initial pH – 2.5, elemental sulfur – 1%). initial pH – 2.5, initial Fe(II) – 3 g/L, elemental sulfur – 1%).
 D. Mishra et al. / Waste Management 28 (2008) 333–338 337

12 3.5. EDAX (energy dispersive X-ray analysis) study

0.5g
10 0.5g(Control) The slow dissolution of lithium in the experiment is the
1g
issue to consider. To prove the slower solubility of lithium
% of Lithium leached

1g (Control)
8
1.5g
than that of cobalt, EDX (energy dispersive X-ray analysis)
1.5g (Control) mapping of the leach residues after leaching was carried out
6 2g (Figs. 10a and b) and it was compared with the original sam-
2g (Control)
ple. The distributions of Co and Li have been indicated in
4 the diagram. The distribution of lithium was observed to
be the same in both leach residues and in the original sample.
2 However, a little distribution of cobalt on the surface of the
leach residues indicated that it has been leached to the solu-
0 tion during the leaching process. The mapping proved that
0 5 10 15
Time in days lithium itself has slow dissolution during the bioleaching
process and there might be some other reasons that inter-
Fig. 9. Lithium leaching at different solid/liquid ratio (w/v). (Conditions:
feres with lithium solubility. Hence, in each set of experi-
initial pH – 2.5, initial Fe(II) – 3 g/L, elemental sulfur – 1%).
ments, cobalt recovery is five times greater than lithium.

cell inhibition at a higher solid/liquid (w/v) ratio, 4. Conclusions

although initial bacterial adaptation was limited to a
solid/liquid ratio of 1 g/100 mL. Earlier, Tuovinen et al.
(1971) reported that cobalt was the most toxic cation
among other heavy metals on the growth of A. ferrooxi-
dans on an elemental sulfur medium. So, when the solid/
liquid ratio was increased, the metal concentration in the
LIBs increased simultaneously, and hence, the cell growth
was arrested by the toxic nature of the metals. At 5 g/L
of solid/liquid ratio, cobalt dissolution was about 65%
and at 10 g/L it was about 56%. Respective control stud-
ies have been carried out and marked as dotted lines in
Figs. 8 and 9.
Results of present work indicated that it is possible to
dissolve metals from spent cathode material of LIBs by
the use of acidophilic bacteria. These cells are able to grow
in the medium containing elemental sulfur and iron as their
energy source. Results revealed that a culture of A. ferroox-
idans can produce sulfuric acid to leach metals indirectly
from the LIBs. Cobalt was leached faster than lithium.
The EDXA mapping demonstrated a slower dissolution
rate of lithium than that of cobalt. The use of Fe(II) ion
in the leaching experiments was to proliferate the cell
growths in the lixiviant: the higher the Fe(II) ion concen-

Fig. 10. (a) SEM image of the raw sample and responding X-ray energy dispersive spectrum element distribution of Li and Co. (b) SEM image of the leach
residues and responding X-ray energy dispersive spectrum element distribution of Li and Co.
338 D. Mishra et al. / Waste Management 28 (2008) 333–338
tration, the slower the metal dissolution. The Fe(III) ion
formed during the course of the investigation was found
to be precipitated with the metals in the leach residues.
Higher solid/liquid ratios prevented bacterial activity in
the process as higher metal concentrations are considered
to be toxic for cells.
Acknowledgements
The authors are thankful to the MOST (Ministry of Sci-
ence and Technology), Korea, for the financial assistance.
First author (D.M.) is thankful to KRF (Korea Research
Foundation) for supporting the fellowship.
References
Brandl, H., Bosshard, R., Wegmann, M., 2001. Computer-munching
microbes: metal leaching from electronic scrap by bacteria and fungi.
Hydrometallurgy 59, 319–326.
Cerruti, C., Curutchet, G., Donati, E., 1998. Bio-dissolution of spent
nickel–cadmium batteries using Thiobacillus ferrooxidans. Journal of
Biotechnology 62, 209–219.
Curutchet, C., Pogliani, C., Donati, E., Tedesco, P., 1992. Effect of iron
(III) and its hydrolysis products (Jarosites) on Thiobacillus ferrooxi-
dans growth and on bacterial leaching. Biotechnology Letters 14, 329–
334.
Ehrlich, H.L., 1981. Geomicrobiology. Marel Dekkar De, New York.
Horvath, B., Gruiz, K., 1996. Impact of metalliferous ore mining activity
on the environment in Gyöngyösoroszi, Hungary. Science of the Total
Environment 184, 215–227.
Lee, C.K., Rhee, K.I., 2002. Preparation of LiCoO2 from spent lithium-
ion batteries. Journal of Power Sources 109, 17–21.
Moore, J.N., Luoma, S.N., 1990. Hazardous wastes from large-scale metal
extraction. Environment Science Technology 24, 1278–1285.
Nakazawa, H., Sato, H., 1995. Bacterial leaching of cobalt-rich ferro-
manganese crusts. International Journal of Mineral Processing 43,
255–265.
Nan, J., Han, D., Zuo, X., 2005. Recovery of metal values from spent
lithium ion batteries with chemical deposition and solvent extraction.
Journal of Power Sources 152, 278–284.
Norris, P.R., 1990. Acidophilic bacteria and their activity in mineral
sulfide oxidation. Microbial Mineral Recovery. McGraw-Hill, New
York.
Olson, G.J., Sakai, C.K., Parks, E.J., Brinckman, F.E., 1990. Bioleaching
of cobalt from smelter wastes by Thiobacillus ferrooxidans. Journal of
Industrial Microbiology 6, 49–52.
Pesic, B., Oliver, D.J., Wichlacz, P., 1989. An electrochemical method to
measuring the rate of ferrous to ferric iron with oxygen in the presence
of Thiobacillus ferrooxidans. Biotechnology and Bioengineering 33,
428–439.
Rohwerder, T., Gehrke, T., Kinzler, K., Sand, W., 2003. Bioleaching
review part A: progress in bioleaching: fundamentals and mechanisms
of bacterial metal sulfide oxidation. Applied Microbiology Biotech-
nology 63, 239–248.
Rossi, G., 1990. Biohydrometallurgy. McGraw-Hill, Hamburg, New
York.
Shin, S.M., Kim, N.H., Sohn, J.S., Yang, D.H., Kim, Y.H., 2005.
Development of a metal recovery process from Li-ion battery wastes.
Hydrometallurgy 79, 172–181.
Silverman, M.P., Lundgren, D.G., 1959. Studies on the chemoautotrophic
iron bacterium Ferrobacillus ferrooxidans. I. An improved medium and
a harvesting procedure for securing high cell yields. Journal of
Bacteriology 77, 642–647.
Stefan, S., Nicole, A.I., 2002. Importance of public relations in recycling
strategies: principles and case studies. Environmental Management 30,
68–76.
Tuovinen, O.H., Niemela, S.I., Gyllenberg, H.G., 1971. Tolerance of
Thiobacillus ferrooxidans to some metals. Antonie van Leeuwenhoek
37, 489–496.
Verstraete, W., 2002. Environmental biotechnology for sustainability.
Journal of Biotechnology 94, 93–100.
Zhang, P., Yokoyama, T., Itabashi, O., Suzuki, T.M., Inoue, K., 1998.
Hydrometallurgical process for recovery of metal values from spent
lithium-ion secondary batteries. Hydrometallurgy 47, 259–271.
Zhu, N., Zhang, L., Li, C., Cai, C., 2003. Recycling of spent nickel–
cadmium batteries based on bioleaching process. Waste Management
23, 703–708.