Minerals Engineering 22 (2009) 311313

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Minerals Engineering
journal homepage: www.elsevier.com/locate/mineng

Technical Note

Recovery of nickel from lateritic nickel ore using Aspergillus niger and
optimization of parameters
S. Mohapatra *, N. Pradhan, S. Mohanty, L.B. Sukla
Institute of Minerals and Materials Technology (CSIR), BMBT, Jaydev Vihar, Bhubaneswar 751013, Orissa, India

a r t i c l e

i n f o

Article history:
Received 12 September 2007
Accepted 12 August 2008
Available online 23 September 2008
Keywords:
Oxide ore
Sulphide ores
Fungal strain
Aspergillus niger
Bioleaching

a b s t r a c t
The present study deals with the extraction of nickel from lateritic nickel ore of Sukinda mines, Orissa,
through microbial leaching using Aspergillus niger. The presence of signicant metals in oxidic nickel ores,
are potential sources of nickel for the future. Thus, experiments were carried out with ore to optimize the
conditions for maximum nickel recovery. Experiments designed as per central composite design technique, were carried out for tting an empirical reduced second order model. The factors studied were
sucrose concentration, pulp density, temperature and duration. In addition to main effects of four factors,
sucrose concentration, pulp density and duration had quadratic effect on the percentage extraction and
there were interactions between different factors also. There was also signicant interaction between
temperature and duration as well as sucrose and pulp density. The F-value for the model shows that
model is signicant at less than a 0.01% level (i.e. at 99.99% condence level). The predicted maximum
nickel extraction was 31.34% with a pulp density of 8.75%, sucrose concentration of 10.04 g/l, temperature 33.8 C and duration of 37.5 days.
2008 Elsevier Ltd. All rights reserved.

1. Introduction
The overburdens generated during chromite mining contains
low concentrations of valuable metals, when exposed to environment for long period, causes serious environmental problems due
to natural weathering mediated by indigenous microbes. The dominating importance of lateritic ores for the future supply of nickel is
about 85% of the total known nickel reserves of the world. (Mohapatra et al., 2007). Recent studies have shown the amenability of
laterite ores to leaching by Aspergillus, Penicillium etc. (Sukla
et al., 1993; Tzeferis and Agatzini-Leonardou, 1994). Organic acids
(oxalic acid, citric acid, gluconic acid etc.) are excreted by the
microorganism into the culture medium as metabolic products
and subsequently dissolve heavy metals by forming salts and
chelates into soluble organo-metallic complexes (Avakyan and
Rabotnova, 1971; Valix et al., 2001).
Central composite design (CCD) technique, a response surface
methodology (RSM), was used to design the present study. RSM,
an experimental strategy for the optimum conditions for a multivariable system, is a much more efcient technique for optimization (Box and Wilson, 1951; Box et al., 1978). In particular, they
are used when several factors have to be studied in order to determine their main effects and interactions.
* Corresponding author. Fax: +91 674 2581637/2581160.
E-mail address: mohapatrasmaranika@yahoo.co.in (S. Mohapatra).
0892-6875/$ - see front matter 2008 Elsevier Ltd. All rights reserved.
doi:10.1016/j.mineng.2008.08.002

The experimental work aims to optimize the bioleaching conditions in shake asks with roasted lateritic nickel ores using Aspergillus niger. Sucrose concentration, pulp density, temperature and
duration of leaching were taken into consideration to evaluate
their effect on the nickel extraction.

2. Materials and methods

2.1. Ore materials
The ore used for the study was nickel laterite obtained from the
major deposits of Sukinda mines, Orissa, India. The ore was sieved
to a mean particle size of +45 lm. Roasting was done at 400 C in a
mufe electric furnace for 5 h in a silica crucible and it converts
goethite (FeOOH) into hematite (Fe2O3). The roasted ore shows
more porous structure and had about two times more surface area
than raw nickel ore (Olanipekum, 2000). A representative sample
was subjected to acid digestion and analyzed by AAS and the ore
contains 1.06% nickel.
A. niger, a laboratory stock culture at IMMT, Bhubaneswar was
tested for feasibility of leaching nickel from low-grade ores. Mineral salt medium (MSM) containing (g/100 ml) NH4NO3 0.3;
KH2PO4 0.1; MgSO4  7H2O 0.05; sucrose 512.5; yeast extract
0.1; pH 6.8, was used for fungal growth. Spore suspension of 5
7 days old culture was used as inoculum.

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S. Mohapatra et al. / Minerals Engineering 22 (2009) 311313

2.2. Experimental design by central composite design

performed with each set of experiments without the addition of

microorganisms.

2.2.1. Shake ask studies

CCD experimental technique was used for carrying out the
shake ask experiments. The CCD is shown in Table 1. CCD contains an imbedded factorial design with center points that is augmented with a group of axial points that allow estimation of
curvature. CCD contains twice as many axial points as there are
factors in the design. The axial points represent the extreme values
(low and high) for each factor in the design (Cochran and Cox,
1957; Montgomery, 1997). In this study, a 24 full factorial central
composite design for four independent variables, each at ve levels
with sixteen factorial points, replicates of four factorial points,
eight axial points, replicates of two axial points and six replicates
at the center points, were employed to t a reduced second order
polynomial model and estimate the experimental error.
A. niger was grown in presence of ore and produces acids to dissolve the mineral, and referred to as indirect leaching. Leaching
experiments were carried out in 250 ml Erlenmeyer asks containing sterilized 100 ml MSM and sterilized adapted ore (ranging between 5% and 20%) with addition of 512.5% sucrose. The pH of the
media was adjusted at 6.8 with 1 N NaOH. Each ask was inoculated with 10 ml of ore adapted spore suspension and incubated
at respective temperatures ranging between 30 C and 40 C in a
rotary shaker at 150 rpm. The incubation periods were 15, 22.5,
30 and 37.5 days. The contents of asks were ltered and the solutions were appropriately diluted and analyzed by AAS to determine
the percentage of nickel leached. Control experiments were also

Table 1
Central composite design (CCD)
Standard
run

Block

A (%)
Sucrose

B (%) Pulp
density

C (C)
Temperature

D (days)
Duration

1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16

Factorial
Factorial
Factorial
Factorial
Factorial
Factorial
Factorial
Factorial
Factorial
Factorial
Factorial
Factorial
Factorial
Factorial
Factorial
Factorial

7.5
12.5
7.5
12.5
7.5
12.5
7.5
12.5
7.5
12.5
7.5
12.5
7.5
12.5
7.5
12.5

8.75
8.75
16.25
16.25
8.75
8.75
16.25
16.25
8.75
8.75
16.25
16.25
8.75
8.75
16.25
16.25

32.5
32.5
32.5
32.5
37.5
37.5
37.5
37.5
32.5
32.5
32.5
32.5
37.5
37.5
37.5
37.5

22.5
22.5
22.5
22.5
22.5
22.5
22.5
22.5
37.5
37.5
37.5
37.5
37.5
37.5
37.5
37.5

17
18
19
20
21
22
23
24

Axial
Axial
Axial
Axial
Axial
Axial
Axial
Axial

5
15
10
10
10
10
10
10

12.5
12.5
5
20
12.5
12.5
12.5
12.5

35
35
35
35
30
40
35
35

30
30
30
30
30
30
15
45

25
26
27
28
29
30

Center
Center
Center
Center
Center
Center

10
10
10
10
10
10

12.5
12.5
12.5
12.5
12.5
12.5

35
35
35
35
35
35

30
30
30
30
30
30

31
32
33
34
35
36

Factorial
Factorial
Axial
Factorial
Axial
Factorial

12.5
7.5
10
7.5
10
7.5

16.25
8.75
12.5
8.75
20
8.75

37.5
37.5
40
32.5
35
37.5

22.5
22.5
30
22.5
30
37.5

3. Results and discussion

3.1. Data analysis
The data generated from shake ask experiments were statistically analyzed to identify the signicant main and interaction effects. Analysis of variance (ANOVA) of the experimental results
was carried out to decide upon the signicant and insignicant effects and thus obtain best possible regression model. It was found
that a reduced quadratic model as given in Eq. (1) could best t
with the experimental data

% Ni Leaching 418:05 11:13  sucrose

0:6792  pulp density
20:40  temperature 2:47  duration
2

 0:6056  sucrose2  0:1278  pulp density

 0:2833  temperature2
2

 0:0173  duration

0:1194  sucrose  pulp density

 0:0334  temperature  duration

The ANOVA for this model is shown in Table 2. The F-value for the
model shows that model is signicant at less than a 0.01% level (i.e.
at 99.99% condence level). Sucrose concentration, pulp density,
temperature and duration have a signicant main and quadratic effect on the nickel leaching. In addition there is signicant interaction between sucrose and pulp density, temperature and duration.
The presence of an interaction (or synergism) indicates that there
is a change in the effect of a factor, when changing the levels of
the other factors i.e. two factors are said to interact if the effect of
a factor depends on the level of another factor.
The R2 value between the experimental and predicted values
was 0.955, which shows that there is good agreement. The lackof-t test shows that the contribution of model error is more signicant than the experimental error for the deviation between
the predicted and experimental values. However, since there is
not much difference between the experimental and predicted values, the model is acceptable. The F-values are all below 0.005, with
the exception of the CD interaction. The maximum, minimum and
average percentage absolute error between the experiment and
predicted were 17.54, 0.43 and 6.48, respectively. Fig. 1 shows
the three-dimensional response surface of nickel extraction (%) as
a function of pulp density (%) and sucrose (%), at optimized temperature (33.8 C) and duration (37.5 days).
The empirical model obtained was used for optimization of the
process parameters to maximize nickel extraction, within the
range in which the experiments were carried out. The optimization
technique used in the present study was Nelder-Mead. Qualitative
and statistical analysis indicates that the extraction of nickel is favored with the increase in sucrose concentrations but it is restricted because of economical point of view. The optimized
conditions obtained from the shake ask experiments were pulp
density 8.75%, sucrose concentration 10.04 g/l, temperature
33.8 C and duration 37.5 days for a maximum nickel extraction
of 31.34%.
A. niger was found to be efcient in extracting nickel at high sucrose concentration in the medium because it serves as the energy
source in the ore as well as for the strain. Drop in pH, was more in
case of high sucrose concentration. Biomass generation was 1.72 g
with high sucrose and 1.05 g dry weight per 100 ml media for low
sucrose concentration. Previous reports suggests that leaching ef-

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S. Mohapatra et al. / Minerals Engineering 22 (2009) 311313

Table 2
ANOVA for response surface reduced quadratic model
Source

Sum of squares

DF

Mean square

F-value

Prob > F

Model
A
B
C
D
A2
B2
C2
D2
AB
CD

1413.965428
44.53987006
737.155338
34.50931626
106.6659618
403.3456429
110.9700402
106.8936004
26.77028073
23.9096887
7.658461371

10
1
1
1
1
1
1
1
1
1
1

141.3965
44.53987
737.1553
34.50932
106.666
403.3456
110.97
106.8936
26.77028
23.90969
7.658461

52.81518
16.63677
275.3462
12.8901
39.84243
150.6598
41.45012
39.92746
9.999377
8.930873
2.860629

<0.0001
0.0004046
<0.0001
0.001407
<0.0001
<0.0001
<0.0001
<0.0001
0.004076
0.006208
0.1032

Signicant

66.9298724
63.99508906
2.934783333

25
14
11

2.677195
4.571078
0.266798

<0.0001

Signicant

Residual
Lack-of-t
Pure error

17.13307

Orissa, India, using A. niger. The experiments were designed using

central composite design technique, to carry out a systematic and
optimized approach to experimentation. The factors studied were
sucrose concentration, pulp density, temperature and duration
and found that in addition to main effects and quadratic effects,
the interactions between different factors also exist. A non-linear
regression model was obtained based on the statistical analysis
and parameters were optimized within the desirable operating
range to maximize nickel extraction. The predicted nickel extraction percentage was 31.34% with a pulp density of 8.75%, sucrose
concentration of 10.04 g/l, temperature 33.8 C and duration of
37.5 days for the experiment.

Acknowledgements

Fig. 1. Three-dimensional response surface curve of nickel extraction (%) showing

the interaction between pulp density and sucrose at various levels (at optimized
duration 37.5 and temperature 33.8 C).

The authors wish to thank the Department of Science and Technology, New Delhi, India, for nancial support. S. Mohapatra
acknowledges CSIR for offering her Senior Research Fellowship.
The authors also acknowledge the Director, Institute of Minerals
and Materials Technology (CSIR), Bhubaneswar, for permission to
publish the paper.

References
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33 C, which is the optimum temperature for the growth of this
organism. At extreme temperatures, the metabolic rate of organism decreases and also the organism might not survive.
4. Conclusions
The present shake ask studies were carried out to optimize the
nickel extraction from roasted nickel laterite of Sukinda mines of

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