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Kinetics of Ferrous Iron Oxidation by Sulfobacillus Thermosulfidooxidans

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Kinetics of Ferrous Iron Oxidation by Sulfobacillus Thermosulfidooxidans Biochemical Engineering Journal 51 (2010) 194–197 Contents lists available at ScienceDirect Biochemical Engineering Journal journal homepage: www.elsevier.com/locate/bej Short communication Kinetics of ferrous iron oxidation by Sulfobacillus thermosulfidooxidans Pablo S. Pina 1, Victor A. Oliveira, Flávio L.S. Cruz, Versiane A. Leão ∗ Universidade Federal de Ouro Preto, Department of Metallurgical and Materials Engineering, Bio&Hidrometallurgy Laboratory, Campus Morro do Cruzeiro, s.n., Ouro Preto, MG 35400-000, Brazil article info abstract Article history: The biological oxidation of ferrous iron is an important sub-process in the bioleaching of metal sul- Received 9 October 2009 fides and other bioprocesses such as the removal of H2S from gases, the desulfurization of coal and the Received in revised form 1 June 2010 treatment of acid mine drainage (AMD). As a consequence, many Fe(II) oxidation kinetics studies have Accepted 10 June 2010 mostly been carried out with mesophilic microorganisms, but only a few with moderately thermophilic microorganisms. In this work, the ferrous iron oxidation kinetics in the presence of Sulfobacillus thermo- sulfidooxidans (DSMZ 9293) was studied. The experiments were carried out in batch mode (2L STR) and Keywords: the effect of the initial ferrous iron concentration (2–20 g L−1) on both the substrate consumption and Growth kinetics Thermophiles bacterial growth rate was assessed. The Monod equation was applied to describe the growth kinetics of −1 −1 Batch processing this microorganism and values of max and Ks of 0.242 h and 0.396 g L , respectively, were achieved. Microbial growth Due to the higher temperature oxidation, potential benefits on leaching kinetics are forecasted. Waste treatment © 2010 Elsevier B.V. All rights reserved. 1. Introduction mesophile microorganism was able to grow in the presence of up to 30gL−1 Fe(II), but the maximum oxidation rate (0.47 g L−1 h−1)was Bioleaching is now a proven technology that has been applied observed only in the presence of 20 g L−1 Fe(II). The thermophiles to the processing of a series of metallic sulfides such as copper, were more sensible to the presence of Fe(II) and did not grow at uranium and refractory gold ores. Two mechanisms have been pro- concentrations higher than 5.8 g L−1 Fe(II). In this substrate concen- posed to explain the bacterial attack on the sulfides: (i) the direct tration, an overall oxidation rate of 0.105 g L−1 h−1 was achieved. mechanism, which relies on the catalytic oxidation of the metal- The authors also determined the specific growth rate () and the lic sulfide. This mechanism is thought to be important whenever yield (Y) for A. brierleyi as 0.028 h−1 and 1.9 × 1010 cells g−1, respec- there is a direct contact between the bacteria and the mineral, as tively. for example, in heap bioleaching. The second mechanism refers to Moderate thermophiles have shown the ability to improve the biological production of Fe(III) and its role on the oxidation bioleaching kinetics of selected sulfides due to the higher bioleach- of sulfides [1] which follows different patterns according to sulfide ing temperature as compared to mesophiles. As the indirect electronic configuration. Some are oxidized directly to sulfate while mechanism plays a key role in most bioleaching systems and only others are oxidized to elemental sulfur. Notwithstanding, ferrous few ferrous iron bio-oxidation kinetics studies were performed iron is a by-product and needs to be biologically re-oxidized by the with moderately thermophilic microorganisms, this work aimed bacteria. to address Fe(II) oxidation with Sulfobacillus thermosulfidooxidans. Owing to the numerous potential applications of ferrous iron bio-oxidation, several works were carried out with mesophilic 2. Materials and methods microorganisms, especially with Acidithiobacillus ferrooxidans and Leptospirillum ferrooxidans [2], and a number of kinetic models have S. thermosulfidooxidans (strain DSMZ 9293) was grown in a −1 −1 been proposed for the oxidation of ferrous iron [3,4] in the temper- medium containing: 0.4 g L (NH4)2SO4, 0.8gL MgSO4·7H2O, −1 −1 −1 ature range usually found in industrial applications. Nemati et al. 0.4gL K2HPO4,10gL of ferrous iron and 0.1 g L yeast extract [3] compared the batch kinetics of Fe(II) oxidation by A. ferrooxi- and provided the inoculum for the bio-oxidation experiments from dans and Acidianus brierleyi, at pH 1.8. The authors observed that the a 48 h grown culture. The bio-oxidation experiments were carried out in batch mode in a bioreactor (New Brunswick Scientific – BioFlo 110) with 2 L ∗ of suspension containing 10% (volume) of inoculum. To produce Corresponding author. Tel.: +55 31 3559 1102; fax: +55 31 3559 1596. the latter, 400 mL of the culture were filtered through a Millipore E-mail addresses: [email protected], [email protected] (V.A. Leão). ␮ 1 Current address: Vale – Centro de Desenvolvimento Mineral, Santa Luzia, Minas (0.22 m) membrane and resuspended in 200 mL of distilled water, Gerais, Brazil. at pH 2.0. Afterwards the pH was reduced to 1.5 and kept at this 1369-703X/$ – see front matter © 2010 Elsevier B.V. All rights reserved. doi:10.1016/j.bej.2010.06.009 P.S. Pina et al. / Biochemical Engineering Journal 51 (2010) 194–197 195 rate dX/dt was calculated according to Eqs. (1) and (2), where is the specific growth rate. 1 dX = (1) X dt dX X = X or Ln = t (2) dt X0 Eq. (2) represents exponential growth and applies to closed systems where growth is the only process affecting cell concentration (X) [7]. A plot of Ln X versus time gives a straight line with slope . Following, a set of specific growth rate values at different ini- tial ferrous iron concentration was achieved and the experimental data were then fit to the Monod equation so that the maximum specific growth rate (max) and the substrate constant (Ks) could be assessed. The Monod equation is the most widely used expres- sion to correlate the growth of bioleaching microorganisms with Fig. 1. Ferrous ion concentrations and cell numbers as a function of time for the ferrous iron concentration and does not consider inhibitory effects −1 ◦ growth of S. thermosulfidooxidans. [Fe(II)]0 =5gL ;50 C, 10% inoculum, pH 1.5. in its standard form [8]. In batch systems, it applies only for bal- anced growth, which is most cultures, occurs concurrently with value in all experiments. A pH meter (Hanna 2221) and glass mem- the exponential growth phase [7]. brane electrode calibrated against pH 4.0 and 7.0 buffers was used Fig. 3 shows the ferrous iron oxidation rate and the microbial for pH measurement. The pH was controlled during the experi- specific growth rate as a function of the initial ferrous iron concen- −1 −1 ments by addition of either concentrate sulfuric acid or sodium tration. A maximum overall oxidation rate of 0.697 g L h was −1 hydroxide. Both the temperature and the stirring rate were main- observed in the experiments carried out with 10 g L Fe(II) and −1 tained constant at 50 ◦C and 31.43 rad s−1, respectively. This latter did not change for higher concentrations (15 and 20 g L )(Fig. 3a). was defined as the value which produced the highest ferrous iron The value achieved for S. thermosulfidooxidans is 5 times higher than oxidation rate. The initial ferrous iron concentration varied from 2 that determined by Nemati and Harrison [6] in experiments carried −1 −1 −1 to 20 g L−1. The solution was aerated at a rate of 1 L min−1 and 5 mL out with A. brierleyi (0.133 g L h , in systems containing 7.5 g L samples were withdrawn regularly and analyzed for ferrous iron Fe(II)) and higher than the value reported by the same authors for −1 −1 −1 concentration and cell number. Cell counts were performed using A. ferrooxidans (0.47 g L h ), at 20 g L Fe(II). a Neubauer chamber and light contrast microscope (Leica). The bacterial specific growth rate shows similar behavior level- −1 −1 The ferrous iron concentration was determined by titration ing out at around 0.25 h for the 10–20 g L Fe(II) concentration −1 with standard potassium dichromate solution in the presence range (Fig. 3b). This value is similar to that (0.35 h ) achieved by Bogdanova et al. [9] in the presence of 9.82 g L−1 FeSO ·7H O, for a ofa1H2SO4:1 H3PO4 solution using an automatic titrator 4 2 (Schott–Tritoline Alpha). All chemicals used in this study were ana- new species of Sulfobacillus with an optimum growth temperature ◦ lytical grade reagents (AR) unless otherwise stated and all solutions of 40 C. Applying the Monod equation to the data here presented were prepared with distilled water. (Fig. 3b), ferrous iron bio-oxidation can be described by the parame- −1 −1 Statistical analysis was carried out using the OriginTM version ters max and Ks as 0.242 h and 0.396 g L , respectively. Nemati 8.0 software to determine the specific growth rate, the Fe(II) oxi- et al. [3] after compiling max and Ks data available in the liter- dation rate as well as the yield values for a 95% confidence interval. ature for the growth of Acidithiobacillus ferooxidans pointed out The data points used to calculate such parameters were those that significant discrepancies in the reported values. Notwithstanding, produced linear regression with correlation coefficients (r2) higher the values observed in the present work are generally higher than than 0.94.
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