Bioresource Technology 97 (2006) 1876–1879

Beneficiation of iron ore slime using Aspergillus niger

and Bacillus circulans
N. Pradhan *, B. Das, C.S. Gahan, R.N. Kar, L.B. Sukla
Regional Research Laboratory, CSIR, Bhubaneshwar 751 013, India

Received 17 December 2004; received in revised form 8 August 2005; accepted 26 August 2005
Available online 13 March 2006

Abstract

Studies were carried out on the removal of alumina from iron ore slime containing (%) Fe2O3 75.7, Al2O3 9.95, SiO2 6.1, Fe (total)
52.94 with the help of Bacillus circulans and Aspergillus niger. B. circulans and A. niger showed 39% and 38% alumina removal after six
and 15 days of in situ leaching at 10% pulp density, respectively. Culture filtrate leaching with A. niger removed 20% alumina at 2% pulp
density with 13 day old culture filtrate. B. circulans was more efficient than A. niger for selective removal of alumina. In case of A. niger
in situ leaching rather than culture filtrate leaching was found to be more effective.
 2005 Elsevier Ltd. All rights reserved.

Keywords: Iron ore slime; Aspergillus niger, Bacillus circulans; Bio-beneficiation; Alumina removal

1. Introduction to time to reduce the gangue constituents so that the bene-

Reserves of high-grade ores are diminishing all over the
world at an alarming rate as a result of rapid increase in de-
mands for metals. The recovery of mineral value from the
low-grade ores using present technology is prohibitively
expensive due to high energy and capital costs. Presently
available physico-chemical methods are not environment
friendly. Bio-beneficiation is considered as eco-friendly,
promising and revolutionary solutions to these problems
and is gaining more importance due to depletion of high-
grade ores and enforcement of strict anti-pollution laws.
Iron ore slimes containing around 48–62% Fe are gener-
ated from the iron ore washing plants and are disposed into
tailing ponds without any further utility. The slimes are not
suitable in iron and steel making due to the extremely finer
size and presence of higher amount of gangue constituents.
Several beneficiation techniques have been tried from time
*
Corresponding author. Tel.: +91 674 2584091 94x528; fax: +91 674
2581636.
E-mail addresses: npradhan@rrlbhu.res.in, nilotpala_pradhan@
yahoo.co.in (N. Pradhan).
ficiated products could be effectively used for iron and steel
making (Das et al., 1992; Pradip, 1994; Prakash et al.,
1999). Gangue minerals present in iron ores comprise both
metallic and non-metallic components. Usually, siliceous
gangue consists of larger proportion of alumina in the form
of clay and laterite along with varying amounts of undesir-
able constituents such as phosphorous, sulphur, titanium,
copper and arsenic. Many microorganisms have been re-
ported to solubilise different alumino-silicate compounds
found in nature. In silicates, silicon is usually surrounded
by four oxygen atoms in tetrahedral fashion (Kretz,
1972) whereas aluminium in aluminosilicates is coordi-
nated with oxygen in tetrahedral or octahedral fashion,
depending upon the mineral (Tan, 1986). In minerals, these
units are arranged in bi- or tri-layers separated by water
layers of variable thickness into which other polar mole-
cules, including some organic molecules can enter. This
type of structure makes them susceptible for weathering
by microorganisms. Si–O bonds of silloxanes linkages
(Si–O–Si) in silicates and aluminosilicates are very strong,
whereas Al–O bonds are somewhat weaker. Thus Si–O
bonds are relatively resistant to acid hydrolysis (Karavaiko

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doi:10.1016/j.biortech.2005.08.010
 N. Pradhan et al. / Bioresource Technology 97 (2006) 1876–1879 1877

et al., 1985), unlike Al–O bonds. Some bacteria and fungi
are known to solubilise silica and silicates. They accom-
plish this by forming chelators, acids or bases, which react
with silicates and exo-polysaccharides, which react with sil-
ica and silicates.
Although the use of different microorganisms in ore
leaching is well-established, use of microorganism to re-
duce alumina and silica from ore has been attempted in
very few investigations (Natarajan and Deo, 2000). This
paper describes the bio-beneficiation of iron ore slimes
using Aspergillus niger and Bacillus circulans.
2. Methods
2.1. Sample
Iron ore slime sample was obtained from Bolani iron ore
washing plant, Orissa, India. Around one ton of the tailing
sample from the washing plant was mixed thoroughly and
representative sample was drawn for the experiments. The
sample was analyzed by wet chemical and instrumental
techniques (Vogel, 1978). The size analysis of the sample
was done in wet conditions with standard sieves. All size
fractions were collected separately, dried in a hot air oven
(105 C) and analyzed by standard methods (Vogel, 1978).
tively. Different sets were incubated for different period
of 5, 10, 15 days, after which the solid residues were sepa-
rated and analyzed as above.
For culture filtrate leaching, A. niger grown in mineral
salt medium for different time durations was used. To the
culture filtrates of different period, iron ore slime at a rate
of 2% pulp density was added and boiled for three hours
with occasional replacement of volume by water. After
boiling, the solid residue was separated by sedimentation
and filtration, dried and analyzed for Al and Fe.
3. Results and discussion
3.1. Chemical and size analysis of iron ore slimes
Complete chemical analysis of Bolani iron ore slime re-
vealed that the sample on an average contained (%) Fe2O3
75.7, Al2O3 9.95, SiO2 6.1, LOI 8.7, Fe (total) 52.94. As
alumina concentration in the sample was quite high, it
could not be used in iron and steel making. Besides this,
the ratio of SiO2:Al2O3 was also very high contradicting
to blast furnace chemistry. The size analysis of the iron
ore slimes down to 20 lm size is shown in Table 1. The re-
sult indicated that the sample was very fine in nature with
76.7% of the material of below 20 lm size.

2.2. Microorganisms 3.2. Beneficiation experiments

B. circulans MTCC 879 and A. niger MTCC 282 were Both Aspergillus sp. and Bacillus sp. are known to be in-
used for the bio-beneficiation of iron ore slime. Strains volved in the leaching and beneficiation processes of silicate
were maintained on nutrient agar and potato dextrose agar ores and minerals (Karavaiko et al., 1980; Avakyan et al.,
slants, respectively. Bromfield medium was used for benefi- 1986). The solubilizing action by microorganisms may in-
ciation studies by B. circulans, which contained (g/L) su- volve the cleavage of Si–O–Si or Al–O framework bonds,
crose 20, yeast extract 1, K2HPO4 0.25, NH4SO4 0.25, or the removal of cations from the crystal lattice of silicate
MgSO4 0.75, sodium bisphosphate 0.30, pH 6.8 ± 0.2. causing the subsequent collapse of silicate lattice structure.
Medium used for growth and beneficiation studies by A. In situ leaching with A. niger varied with incubation
niger was mineral salt medium containing (g/L) Sucrose time and 5, 10 and 15 days incubation resulted in 10%,
10, K2HPO4 1, NH4NO3 3, MgSO4 0.5, pH 6.8 ± 0.2. 27% and 38% removal of alumina, respectively (Table 2).
Final pH of the medium was 2.1, 1.75 and 1.32 after 5,
2.3. Beneficiation process 10 and 15 days, respectively. About 2% and 25% iron was
also leached out along with the alumina in 10 and 15 days
In situ leaching experiments with B. circulans were car- of leaching, respectively. Removal of alumina was maxi-
ried out in 100 ml of Bromfield medium in 250 ml Erlen- mum with 15 days incubation but simultaneous dissolution
meyer flask under sterile conditions at a pulp density, of Fe was also substantially high.
temperature and agitation speed of 10%, 35 C and
150 rpm, respectively. Inoculation was done with 10% inoc- Table 1
ulum (v/v) containing 1 · 108 cfu/ml of B. circulans and Size analysis and distribution of Al2O3 and Fe in different size fractions
incubation time was six days. At the end of experiments, Size (lm) Weight (%) Fe (%) Al2O3 (%)
solid residue was separated by filtration through medium
+212 0.6 54.45 4.97
fast filter paper, dried in hot air oven and analyzed for 212 + 150 0.4 58.64 2.87
Al and Fe. 150 + 105 3.2 59.76 1.95
With A. niger, in situ leaching was done in 100 ml of 105 + 75 5.7 62.55 2.66
mineral salt medium under sterile conditions. Iron ore 75 + 45 9.1 62.00 2.50
slime sample was added at 10% pulp density (w/v). Inocu- 45 + 30 2.7 62.55 2.98
30 + 20 1.6 64.50 3.19
lation was done with 10% inoculum (v/v) containing 20 76.7 50.27 4.73
1 · 106 spores/ml of A. niger spores. Incubation tempera-
Head 100 52.98 9.95
ture and agitation speed were 35 C and 150 rpm, respec-
1878 N. Pradhan et al. / Bioresource Technology 97 (2006) 1876–1879

Table 2
In situ leaching with A. niger and B. circulans
Microorganism Duration (days/pH) Al2O3 (%) Fe (%) Al2O3 removed (%) Fe removed (%)
Original sample – 9.56 53.62 – –
A. niger 5/2.10 8.63 53.12 10 0.93
A. niger 10/1.75 7.01 52.49 27 2.10
A. niger 15/1.32 5.93 40.21 38 25.0
B. circulans 6/4.03 5.80 51.74 39 3.50

Table 3
Culture filtrate leaching with A. niger
pH of culture filtrate Culture age (days) Strength of acid (Moles) Fe (T) (%) Al2O3 (%) Al2O3 removed (%)
1.35 13 0.18 53.00 7.64 20
2.10 6 0.07 51.76 8.10 15
3.02 2 0.03 52.44 8.78 8

For culture filtrate leaching A. niger grown in mineral
salt medium of initial pH of 6.8 was used, which after 2,
6 and 13 days growth resulted in pH around 3, 2 and 1,
respectively. Beneficiation studies with A. niger using cul-
ture filtrate of different pH showed that pH 1 was more
suitable with 20% removal of alumina (Table 3). At pH 2
and 3, the removal of alumina was 15% and 8%, respec-
tively. Some amount of iron was also leached out along
with alumina but was below 4%. Higher leaching of alu-
mina at lower pH could be due to higher acidity and more
metabolites of the culture filtrate (Table 3). Alumina re-
moval was better in case of in situ leaching compared to
culture filtrate leaching.
During metabolism, microorganisms convert glucose or
other carbohydrates into variety of products, including or-
ganic acids. Bio-leaching processes are mediated due to the
chemical attack by the extracted organic acids on the ores.
Acids usually have dual effect of increasing metal dissolu-
tion by lowering the pH and increasing the load of soluble
metals by complexing/chelating into soluble organo-metal-
lic complexes (Burgstaller and Schinner, 1993). Microbially
produced ligands of divalent cations have been shown to
cause dissolution of calcium containing silicates. These
ligands include 2-ketogluconic acid (Duff et al., 1963),
gluconic acid (Kim et al., 1997; Fenice et al., 2000;
Vandevivere et al., 1994), citric (Pandey et al., 2000), oxa-
lic, humic acid (Bennett et al., 1988) etc. A wide variety of
microorganisms have been reported to produce acidic
metabolites, which are responsible in solubilisation of alu-
mino-silicates (Karavaiko et al., 1979). Dissolution of
spodumene (LiAlSi2O6) has been reported by many micro-
organisms including A. niger (Karavaiko et al., 1980;
Avakyan et al., 1986) by production of acid. Alkaline con-
ditions have been reported to be very conducive to mobilize
silicon, from silicates, aluminosilicates or even quartz
(Karavaiko et al., 1985; Kutuzova, 1969). But in our case
we find that alumina removal by A. niger was associated
with increase in acidity of the medium due to production
of acidic metabolites.
In situ leaching with B. circulans resulted in 39% re-
moval of alumina with simultaneous removal of only
3.5% iron in six days (Table 2). Drop in pH was from 6.8
to 4.03, which was not significant when compared to A.
niger. Bacillus sp. is known to produce extra-cellular poly-
saccharides (Shih et al., 2001) and other metabolites, which
may be playing a role in alumina solubilisation from iron
ore slime. Extra-cellular polysaccharides have been re-
ported to be involved in silicon release (Avakyan et al.,
1986). Such polysaccharides, of bacterial or fungal origin,
are able to react with silioxanes to form organic silioxane.
Bacillus mucilaginosus (Malinovskaya et al., 1990) was
effective in solubilising Si in addition to Li and Al by reac-
tion of its extra-cellular polysaccharides with the silicate of
spodumene. In case of B. circulans, drop in pH was less
when compared to A. niger but solubilization of Al to sig-
nificant extent even at pH 4.03 indicated the role of extra-
cellular polysaccharides. Culture filtrate leaching and
in situ leaching with these cultures indicated that it was
possible to remove the silica and alumina from iron ore
slime with the help of microorganisms. Simultaneous disso-
lution of Fe was less with B. circulans as compared to A.
niger which was desirable.
4. Conclusions
Bolani iron ore slime containing 53% Fe, 9.56% Al2O3
and 6.14% SiO2 was subjected to bio-beneficiation studies.
In situ leaching by A. niger removed about 38% of alumina
in 15 days, while B. circulans could removed 39% of
alumina after six days at 10% pulp density. Culture filtrate
leaching with A. niger removed 20% alumina with 13 day
culture filtrate at 2% pulp density. With B. circulans simul-
taneous dissolution of Fe was also less. Thus, B. circulans
proved to be more effective than the A. niger in removal
of alumina from the iron ore slime.
Acknowledgements
Authors are grateful to Dr. R.P. Das, Director, Regio-
nal Research Laboratory, Bhubaneshwar for his kind per-
mission to publish this paper.
 N. Pradhan et al. / Bioresource Technology 97 (2006) 1876–1879
References
Avakyan, Z.A., Pivovarora, T.A., Karavaiko, G.I., 1986. Properties
of a new species, Bacillus mucilaginosus. Mikrobiologiya 55, 369–
374.
Bennett, P.C., Melcer, M.E., Sigel, D.I., Hassett, J.P., 1988. The
dissolution of quartz in dilute aqueous solutions of organic acids at
25 C. Geochimica Cosmochimica Acta 52, 1521–1530.
Burgstaller, W., Schinner, E., 1993. In: Torma, A.E., Wey, J.E.,
Askshmanan, V.L. (Eds.), Metal Leaching with Fungi in Biohydro-
metallurgical Techniques. The Mineral, Metal and Materials Society,
Warrendale, PA, pp. 325–333.
Das, B., Prakash, S., Mohapatra, B.K., Bhaumik, S.K., Narsimhan, K.S.,
1992. Beneficiation of iron ore slimes using hydrocyclone. Minerals
and Metallurgical Processing 4, 101–103.
Duff, R.B., Webley, D.M., Scott, R.O., 1963. Solubilization of minerals
by 2-ketogluconic acid producing bacteria. Soil Science 95, 105–
114.
Fenice, M., Selbman, L., Federici, F., Vassilev, N., 2000. Application of
encapsulated Penicillium variable in solubilization of rock phosphate.
Bioresource Technology 73, 157–162.
Karavaiko, G.I., Avakyan, Z.A., Krutsko, V.S., Zhdanov, A.V., Pisku-
nov, V.P., 1979. Microbiological investigations on a spodumene
deposit. Mikrobiologiya 48, 383–398.
Karavaiko, G.I., Belkanova, N.P., Eroshchev-Shak, V.A., Avakyan, Z.A.,
1985. Role of microorganisms and some physicochemical factors of the
medium in quartz destruction. Mikrobiologiya 53, 795–800.
Karavaiko, G.I., Krutsko, V.S., MelÕnikova, E.O., Avakyan, Z.A.,
Ostroushko, Yu.I., 1980. Role of microorganisms in spodumene
degradation. Mikrobiologiya 49, 402–406.
Kim, K.Y., Jordan, D., Krishnan, H.B., 1997. Rahnella aquatilis, a
bacterium isolated from soybean rhizosphere, can solubilize hydroxy-
apatite. FEMS Microbiology Letters 153, 273–277.
1879
Kretz, R., 1972. Silicon: elements and geochemistry. In: Fairbridge, R.W.
(Ed.), The Encyclopedia of Geochemistry and Environmental Sciences,
Encyclopedia of earth sciences series, vol. IVA. Van Norstrand
Reinhold, New York, pp. 1091–1092.
Kutuzova, R.S., 1969. Release of silica from minerals as result of
microbiological activity. Mickrobiologiya 38, 596–602.
Malinovskaya, I.M., Kosenko, L.V., Vostelko, S.K., Podgorskii, V.S.,
1990. Role of Bacillus mucilaginosus polysaccharide in degradation of
silicate minerals. Mikobiologiya 59, 49–55.
Natarajan, K., Deo, N., 2000. Role of bacterial interaction and
bioreagents in iron ore floatation. International Journal of Mineral
Processing 62, 143–157.
Pandey, A., Soccol, C.R., Nigam, P., Soccol, V.T., Vandenberghe, L.P.S.,
Mohan, R., 2000. Biotechnological potential of agro-industrial
residues II: Cassava bagasse. Bioresource Technology 74, 81–87.
Pradip, P., 1994. Beneficiation of alumina rich Indian iron ore slimes.
Metal Material and Processes 6, 170–194.
Prakash, S., Das, B., Mohanty, J.K., Venugopal, R., 1999. Recovery of
iron minerals from quartz and corundum mixture by selective coating.
International Journal of Mineral Processing 57, 87–103.
Shih, I.L., Van, Y.T., Yeh, L.C., Lin, H.G., Chang, Y.N., 2001.
Production of a biopolymer flocculant from Bacillus licheniformis
and its flocculation properties. Bioresource Technology 78, 267–272.
Tan, K.H., 1986. Degradation of soil minerals by organic acids. In: Haung
P.M., Schnitzer, M. (Eds.), Interaction of soil minerals with natural
organics and microbes. SSSA Special publication Number 17. Soil
Science Society of America Madison, WI, pp. 1–27.
Vandevivere, P., Welch, S.A., Ullman, W.J., Kirchman, D.L., 1994.
Enhanced dissolution of silicate minerals by bacteria at near neutral
pH. Microbial Ecology 27, 241–251.
Vogel, A.I., 1978. Textbook of Quantitative Inorganic Analysis Including
Elementary Instrumental Analysis. ELBS and Longman publications,
London.