World J Microbiol Biotechnol (2009) 25:101106

DOI 10.1007/s11274-008-9866-4

ORIGINAL PAPER

Are bioleaching rates determined by the available particle surface

area concentration?
Pedro Valencia Fernando Acevedo

Received: 28 April 2008 / Accepted: 22 September 2008 / Published online: 7 October 2008
Springer Science+Business Media B.V. 2008

Abstract It is well known that pulp density and particle

size determine the available surface area concentration and
have an influence in the overall rate of bioleaching of
minerals. As metal solubilization takes place through the
surface area of the particles, it can be expected that different combinations of pulp densities and particle sizes
giving the same surface area concentration would determine the same leaching rate. The objective of this work
was to test this hypothesis on the effect of surface area
concentration, pulp density and particle size of the biooxidation of a pyritic gold concentrate by the thermophilic
Archaeon Sulfolobus metallicus in shake flasks. The gold
concentrate was used at 2.5%, 5%, 10%, and 15% w/v pulp
density and at four size fractions: 150106, 10675, 7538
and 38 lm. Temperature was 68C and the initial pH was
2.0. Results showed that the volumetric productivities of
iron and sulfate depend not only on the surface area concentration but also on pulp density and particle size
considered separately. These two variables not only
determine surface area but also exert additional effects on
the process, so the hypothesis was not confirmed. Maximum attained iron productivity was 1.042 g/l day with the
7538 lm fraction at 5% pulp density. Maximum sulfate
productivity was 4.279 g/l day with the 7538 lm fraction
at 10% pulp density.
Keywords Bioleaching
Iron volumetric productivity

Particle size
Pulp density
Sulfate volumetric
productivity
Surface area concentration
Thermophilic
Archaeon
P. Valencia (&)
F. Acevedo
School of Biochemical Engineering, Catholic University of
Valparaiso, Av. Brasil 2147, Valparaiso, Chile
e-mail: pvalencia@vtr.net

Introduction
The bioleaching of minerals and the biooxidation of
refractory gold concentrates take place through the surface
of the mineral particles and its overall rate is influenced by
several operational factors. It is well known that increasing
pulp densities and decreasing particle sizes have positive
effects in the rate of biooxidation, a fact that can be
interpreted as a result of an increasing surface area concentration. Several investigators have also noted that these
factors together with others such as mechanical effects,
metabolic stress caused by uncomfortable environmental
conditions and inhibitory concentrations of ferric ion and
heavy metals can lead to a negative overall effect (Torma
et al. 1972; Groudev 1986; Nemati and Harrison 1999;
Nemati et al. 2000; Sissing and Harrison 2003; Acevedo
et al. 2004).
The effects of high pulp densities and small particle
sizes have been studied in bioleaching with mesophilic
bacteria for a long time (Torma et al. 1972; Groudev
1986). When working with thermophilic Archaea, the
negative effect of high solids concentration is likely to be
larger because of their weaker cell wall (Kandler and
Konig 1998) that makes them susceptible to mechanical
damage and metabolic stress (Norris 1997; Toma et al.
1991; Rikmanis et al. 2007). Decreasing particle sizes can
reduce bioleaching rate for a number of reasons, i.e., difficulties in cell attachment and increased rate of collision
(Nemati et al. 2000; Howard and Crundwell 1999; Harrison et al. 2003).
Irrespectively of these positive and negative effects, an
important question arises: Are bioleaching rates determined solely by the available surface area concentration as
is the case in heterogeneous chemical reactions (Levenspiel
1972). Or could it be that different results are obtained with

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World J Microbiol Biotechnol (2009) 25:101106

different combinations of pulp densities and particles sizes

giving the same surface area?
The objective of this work was to test the hypothesis that
the surface area concentration by itself determines the rate
of biooxidation of a pyritic gold concentrate by the thermophilic Archaeon Sulfolobus metallicus in shake flasks.

Materials and methods

densities were 2.5%, 5%, 10%, and 15% w/v for each
concentrate fraction. Cultures were performed in 500-ml
Erlenmeyer flasks with 100 ml of suspension of concentrate in culture medium and were inoculated with 10 ml of
active culture with 10 g of total soluble iron per liter.
Samples of 1.5 ml were taken each 4 days and centrifuged
at 10,000g for 10 min and the clear liquid used for performing analyses. Reported results correspond to single
experiments run after repeated successive batches until
constant behavior was obtained.

Microorganism and culture medium

Analytical methods
A strain of S. metallicus kindly supplied by Dr. A. Ballester (Universidad Complutense, Madrid) was used
throughout this work. Norris medium (Norris 1989) was
used throughout (per liter 0.4 g ammonium sulfate, 0.5 g
magnesium sulfate, 0.2 g monoacid potassium phosphate,
0.1 g potassium chloride). A flotation gold concentrate was
the energy source. Initial pH was adjusted to 2.0 with
sulfuric acid.
Mineral concentrate and particle size distribution
A pyritic gold flotation concentrate supplied by Refinera
Las Ventanas (Las Ventanas, V Region, Chile) was used. It
contained 15 g Au/tonne and 67.6% pyrite, the rest being
minor quantities of copper sulfide species and 22.2%
gangue. The concentrate was screened and classified into
four size fractions: 150106, 10675, 7538 and 38 lm.
Each fraction presented minor differences in composition
as shown in Table 1.
The determination of the particle size distribution of
each fraction was done by means of digital photographs of
dispersions over microscope slides and computerized
analysis of the images.

Ferrous ion was determined by a modified Muir method

(Herrera et al. 1989) based on the formation of a red
complex with 1,10-phenanthroline; the color was measured
by spectrophotometry at 510 nm. Sodium fluoride was
used to avoid ferric ion interference. Total soluble iron was
measured by reduction of the ferric ion with hydroxylamine chloride and quantification of the ferrous ion by the
above mentioned method. Ferric ion was calculated by
difference.
Sulfate was quantified by precipitation volumetry using
kit 05542-23 of Cole-Parmer (Cole-Parmer Instrument Co.,
Vernon Hills, IL). Sulfate ions are precipitated with BaCl2
and the excess barium ions determined by titration.
Determinations of the maximum volumetric
productivity
Maximum iron and sulfate volumetric productivities (QP)
were determined from the curves of solubilization kinetics
as the slope of the straight line tangent to the curve drawn
from the pseudo-origin (0, S0), where S0 is the initial species concentration. Iron productivities were calculated from
total soluble iron data.

Biooxidation experiments
Batch biooxidation runs were carried on for 40 days in a
rotary shaker (Environmental Incubator Shaker, New
Brunswick Scientific Co.) at 68C and 220 rev min-1. Pulp

Table 1 Composition of the concentrate and of each size fraction

Size fraction
(lm)

Pyrite
(%)

Fe
(%)

Total
150

67.55
72.13

31.48 36.07 15.00

35.90 43.40 31.31

22.20

150106

72.50

35.50 44.24 22.00

21.60

10675

76.79

37.80 47.05 19.70

16.46

7538

74.22

37.00 43.73 28.40

17.48

38

58.85

30.39 34.90 65.50

30.81

123

S
(%)

Au (g/
tonne)

Gangue
(%)

Results and discussion

Surface area concentration was calculated from the results
of particle size distribution determinations assuming
spherical particles. First the total surface was obtained for
each particle size and each pulp density, and then the
surface area concentration was calculated dividing those
values by the liquid medium volume. These results are
shown in Table 2.
Typical iron solubilization kinetics is shown in Fig. 1,
from which volumetric productivities were calculated.
Each pair of factors, particle size and pulp density, was
associated with a maximum volumetric productivity. It can
be seen that the Fe2? concentration was low at all times
and particle sizes and that the Fe3?/Fe2? ratio increased

World J Microbiol Biotechnol (2009) 25:101106

103

with decreasing particle sizes. The same behavior was

observed with decreasing pulp densities (results not
shown). Figure 2 depicts these productivities for iron

solubilization as a function of surface area concentration

with pulp density as a parameter. For each pulp density the
surface area was varied by changing the particle size in the
range shown in Table 1. The increase in surface area
produced an increase in productivity up to a maximal
value, except for the case of 2.5% w/v pulp density.
The maximum iron volumetric productivities at different
surface areas with particle size as parameter are shown in
Fig. 3; this time the different surface areas were obtained
varying the pulp density. Again, the increase in surface
area was accompanied by an increase in productivity up to
a certain value that depended on the particle size. An
exception to this behavior was the run with the larger
particle size (150106 lm). Table 3 is a general summary
of the results for iron productivity. It can be noted that,

Table 2 Surface area concentration as a function of particle size and

pulp density
Fraction (lm)

Surface area concentration (m2/l)

Pulp density (% w/v)
2.5%

5.0%

150106

0.54

1.09

2.18

10675

1.39

2.79

5.57

8.36

7538

1.72

3.43

6.87

10.30

38

3.50

7.00

13.99

20.99

15%
3.26

20

20
150-106 m

Concentration (g/l)

18

106-75 m

18

16

16

14

14

12

12

10

10

0
0

10

15

20

25

t (days)

10

15

20

25

20

25

t (days)

20

20
75-38 m

18

Concentration (g/l)

Fig. 1 Kinetics of iron

solubilization by Sulfolobus
metallicus in shake flasks.
Experimental conditions: 5%
w/v pulp density, initial pH 2.0,
68C, 220 rev min-1. j total
iron concentration; m Fe3?;
d Fe2?

10%

-38 m

18

16

16

14

14

12

12

10

10

0
0

10

15

t (days)

20

25

10

15

t (days)

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World J Microbiol Biotechnol (2009) 25:101106

1.2

Table 3 Maximum volumetric iron productivity as a function of

particle size and pulp density

1.0

Fraction (lm) Maximum volumetric iron productivity (g Fe/l day)

QP (g Fe /lday)

Pulp density (% w/v)

2.5%

5.0%

10%

15%

150106

0.776

0.646

0.420

0.084

10675

0.712

0.985

0.794

0.318

7538

0.740

1.042

1.089

0.582

38

0.819

0.948

1.009

0.618

0.8

0.6

0.4

0.2

4
0.0
0

10

15

20

25

1.2

-2

Fig. 2 Effect of the surface area concentration on the maximum

volumetric iron productivity (QP) in shake flasks at different pulp
densities. Experimental conditions: 68C, initial pH 2.0 and
220 rev min-1. Pulp densities, % w/v: j 2.5; d 5.0; m 10.0; . 15.0

QP (g SO4 /lday)

Surface Area Concentration (m /l)

1
1.0

QP (g Fe /lday)

0
0.8

10

15

20
2

Surface Area Concentration (m /l)

0.6

Fig. 4 Effect of the surface area concentration on the maximum

volumetric sulfate productivity (QP) in shake flasks at different pulp
densities. Experimental conditions: 68C, initial pH 2.0 and
220 rev min-1. Pulp densities, % w/v: j 2.5; d 5.0; m 10.0; . 15.0

0.4

0.2

0.0
0

10

15

20
2

Surface Area Concentration (m /l)

Fig. 3 Effect of the surface area concentration on the maximum
volumetric iron productivity (QP) in shake flasks at different particle
size fractions. Experimental conditions: 68C, initial pH 2.0 and
220 rev min-1. Particle sizes, lm: j 38; d 7538; m 10675;
. 150106

except for the larger particle size fraction, the productivity

presented a maximum with respect to pulp density.
Maximum sulfate productivities at different particle
sizes and pulp densities associated to surface area concentrations are presented in Figs. 4 and 5. These results are
analogous to those of iron productivity and are summarized
in Table 4. It can be noted that maximum values are

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obtained at intermediate conditions of pulp density and

particles. Similarly than in the case of iron productivity the
larger fraction (150106 lm) was an exception to this rule.
The ratio between sulfate and soluble iron at the end of the
process is very near what is to be expected from pyrite
composition. Transient deviations during the biooxidation
may be due to the formation of different sulfur species
intermediates.
The results for iron and sulfate productivities show that
the bioleaching of pyrite by S. metallicus depended not
only on the surface area but also on solids concentration
and particle size, probably because these two variables
provoke different effects on the bioleaching rate. No
increase in iron and sulfate productivities was observed as
a result of increasing surface areas obtained by decreasing
particle size from the 7538 lm to the 38 lm fraction.
These results suggest the existence of an additional factor
affecting the biooxidation process. This effect has been

World J Microbiol Biotechnol (2009) 25:101106

105

surface area concentration is the sole determinant of the

biooxidation rate, which depends also on pulp density and
particle size as separate variables.

Acknowledgements
project 1020768.

References

This work was supported by FONDECYT

-2

QP (g SO4 /lday)

0
0

10

15

20
2

Surface Area Concentration (m /l)

Fig. 5 Effect of the surface area concentration on the maximum
volumetric sulfate productivity (QP) in shake flasks at different
particle size fractions. Experimental conditions: 68C, initial pH 2.0
and 220 rev min-1. Particle sizes, lm: j 38; d 7538; m 10675;
. 150106
Table 4 Maximum volumetric sulfate productivity as a function of
particle size and pulp density
Fraction (lm)

Maximum volumetric sulfate productivity

(g SO2-/l day)
Pulp density (% w/v)
2.5%

5.0%

10%

15%

150106

2.775

2.419

1.647

0.521

10675

2.891

3.494

2.900

1.364

7538

2.900

3.556

4.279

2.329

38

2.650

2.977

3.521

2.410

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The results obtained in our work are similar, but we do
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particle sizes. Rather we think that difficulties in cell
attachment to so small particles together with metabolic
stress could be the main reasons for this behavior.
Finally, it can be concluded that the observations
reported in this work reject the hypothesis by which the

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