Biotechnology Letters, Vol 20, No 3, March 1998, pp.

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CO2 supply in the biooxidation of an enargite-pyrite gold concentrate

F. Acevedo*, J.C. Gentina and N. Garc a
School of Biochemical Engineering, Universidad Catolica de Valpara so, Casilla 4059 ,Valpara so, Chile CO2 supply at 4% (v/v) in air increased the biooxidation of a gold concentrate (41% enargite, 43% pyrite) with Thiobacillus ferrooxidans in a 2-l bioreactor at 4% (w/v) solids concentration, 35C and pH 2.4. Extraction increased from 21 to 69% for Fe, 19 to 25% for As, and 16 to 19% for Cu. Suspended biomass increased from 2.6107 to 1.2108 cells/ml. It is concluded that the biooxidation of this gold concentrate is limited by the rate of CO2 supply.

Introduction In the bioleaching of minerals with chemoautotrophic bacteria, the carbon source, CO2, is usually supplied by air. Because of the very low CO2 content of air (0.03% v/v), the growth of the bacteria may become CO2 limited if the aeration is set to satisfy the oxygen demand. In fact, it is thought that CO2 limitation is more likely to occur than oxygen limitation (Acevedo and Gentina, 1989 ; Boogerd et al., 1990). If that is the case, CO2 enrichment of the air should cause an increase in the growth, and consequently, in the leaching rate. CO2 limitation in Thiobacillus ferrooxidans growth and bioleaching ability have been analysed and experimentally conrmed by several authors (Nagpal et al., 1993). Nevertheless, the available information on this topic is not easy to compare because of differences in mineral composition, bacterial strains and experimental conditions. Moreover, in many cases data on mass transfer capabilities of the equipment involved are not reported. The carbon dioxide limitation is relieved at some critical value that varies from one condition to another. It has also been found that high CO2 concentrations are inhibitory (Nagpal et al., 1993). The large scale biooxidation of gold concentrates has become a reality in the past few years (Brierley, 1995). Most of the work done in this area have been performed with arsenopyrite and pyrite minerals, but no more than three papers can be cited on the biooxidation of enargite, Cu3AsS4 (Escobar et al., 1997). The objective of this work was to test the hypothesis of carbon dioxide limitation when the aeration rate is based on the oxygen demand and to determine the optimum CO2 enrichment in the biooxidation of a gold concentrate of high enargite and pyrite content.
1998 Chapman & Hall

Table 1 Composition (w/w) of the enargite-pyrite gold concentrate.

Au g/g 42.0 Cu % 21.1 Fe % 22.6 S % 37.8 As % 7.7 Cu3AsS4 % 40.7 FeS2 CuFeS2 % % 42.8 3.9

Experimental Microorganism and mineral Strain R-2 of Thiobacillus ferrooxidans, supplied by the Laboratory of Microbiology, Faculty of Biological Sciences, Universidad Cat olica de Chile, Santiago, was grown in a 9K-type medium (Silverman and Lundgren, 1959), with ferrous sulphate replaced by 4% (w/v) gold concentrate as the energy source. The mineral concentrate was kindly supplied by Minera El Indio (La Serena, Chile), and had the composition detailed in Table 1. Equipment and operating conditions Experiments were performed batchwise in a 2-l fermenter operated with 1.3 l medium. Agitation at 500 rpm was provided by two six-bladed disc turbines. The temperature was controlled at 35C and the pH at 2.4. The aeration rate was 1.2 vvm. Air was supplemented with 1, 2, 4, or 6% v/v CO2. A control run gassed with regular air (0.03% CO2) was also performed. The Eh was measured with a combination probe with a Ag/AgCl reference electrode. Determination of the volumetric CO2 mass transfer coefcient kLa for CO2 was determined by the dynamic method of Andr e et al. (1981), following the response of the CO2 concentration in the exit gas to a step change in ow rate. It was also calculated from kLa measurements for oxygen by the stationary method described by Sobotka et al. (1982), considering that (Nagpal et al.,1993):
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F. Acevedo et al. kLa]C kLa]O (DC/DO)2/3 where kLa]C and kLa]O are the mass transfer coefcients for CO2 and O2, and DC and DO are the diffusivities of CO2 and O2 in water, respectively. Analysis Previous experiments showed that in this system, after the rst few days of operation, the cell population is distributed roughly 50% attached to the mineral and 50% suspended in the liquid. Suspended cell concentration was determined by direct counting in a Petroff-Hausser chamber and by alkaline hydrolysis and Lowry protein, assuming an average cell composition of 37% protein. Ferrous ion was quantied by a modied phenanthroline method (Herrera et al., 1989); ferric iron was reduced with hydroxylamine hydrochloride and then assayed with phenanthroline. Copper was analysed by iodometry (Flashka et al., 1969), and arsenic by reduction, distillation and iodometry (Brescia et al., 1970). CO2 in the exit gas was measured with a Beckman infrared analyser. Dissolved oxygen was monitored with a galvanic probe. Results and discussion The results show a marked effect of carbon dioxide concentration on the kinetics of growth and bioleaching. Typical kinetic patterns are shown in Figures 1 and 2 for the case of 4% CO2. Very slow growth can be appreciated after a nine-day period of fast growth (Figure 1). Because at that time all nutrients were in excess, a longer growth period could be expected; the reason for this behaviour is not clear at this time, but it may be a combined effect of inhibition by Fe3 and other ions and cell adhesion to precipitated jarosite.

Figure 2 Kinetics of iron and copper solubilisation in the bioleaching of an enargite-pyrite gold concentrate by Thiobacillus ferrooxidans in a 2-l agitated tank at 4% pulp density, 35C, pH 2.4, 500 rpm,1.2 vvm, 4% CO2.

Soluble iron accumulated in the leachate up to 5.9 g/L at day 12 and then declined due to Fe3 precipitation (Figure 2). Almost all of the iron in the liquid was present as ferric ion, denoting a very efcient bacterial oxidation of the ferrous ion. Although the copper curve apparently does not suggest it, some precipitation must occur because 1.8 g Cu/l is less than the copper that stoichiometrically corresponds to the 0.79 g As/l present in the liquid. Furthermore, some copper should also be supplied by chalcopyrite and other minor species present in the concentrate. The suspended cell population also increased signicantly from 2.6107 cells/ml in the case of regular air to 1.2108 cells/ml in the run with 4% CO2. At 6% CO2 the nal population dropped to 7107. In all runs, the Eh increased from initial values of 250280 mV to steady gures of 500505 mV after 5 to 8 days of leaching. Table 2 presents a summary of the results of the ve runs. The specic growth rates and the extraction rates of Fe, Cu, and As exhibit a maximum at 4% CO2 in the inlet gas.
Table 2 Effect of CO2 enrichment on the maximum specic growth rate and leach rates of Fe, Cu and As after 24 days of biooxidation.
1 (h1) air 1% 2% 4% 6%
1

Fe2 (mg/lh) 5.00 5.03 17.00 20.00 3.65

Cu (mg/lh) 2.82 2.84 3.77 3.93 3.58

As (mg/lh) 1.04 1.04 1.28 1.35 1.14

Figure 1 Kinetics of cell growth and arsenic solubilisation in the bioleaching of an enargite-pyrite gold concentrate by Thiobacillus ferrooxidans in a 2-l agitated tank at 4% pulp density, 35C, pH 2.4, 500 rpm, 1.2 vvm, 4% CO2.

CO2 CO2 CO2 CO2

0.012 0.015 0.022 0.027 0.019

Calculated from cell count data, 2 calculated at the time of maximum concentration before precipitation.

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CO2 supply in the biooxidation of an enargite-pyrite gold concentrate

Table 3 Effect of CO2 enrichment in the extraction of Fe, Cu, and As after 24 days of biooxidation.
Fe (%) air 1% 2% 4% 6% 21.23 34.23 59.35 69.38 20.29 Cu (%) 16.17 18.60 18.48 19.28 17.59 As (%) 19.39 19.39 23.91 25.24 21.29

CO2 CO2 CO2 CO2

In conclusion, the results conrmed the carbon dioxide limitation in the biooxidation of this enargite-pyrite concentrate in a 2-l tank bioreactor operated at 35C, pH 2.4, 500 rpm and 1.2 vvm. The optimal carbon dioxide concentration in the inlet gas was 4%. The limitation resulted from the combination of two factors: the low concentration of CO2 in air and an insufcient ow rate. Acknowledgements The authors wish to acknowledge support received from FONDECYT-Chile, project 1950974 and from DGIPUCV, project 203.780/94. References
Acevedo, F and Gentina, JC (1989). Bioprocess Eng 4: 223229 Andr e, G, Moo-Young, M and Robinson, CW (1981). Biotechnol Bioeng 23: 11611122 Boogerd, FC, Bos, P, Kuenen, JG, Heijnen, JJ and van der Lans, RG (1990). Biotechnol Bioeng 35: 11111119 Brescia, F, Arents, J, Meilich, H and Turk, A (1970). M etodos de Laboratorio Qu mico: Fundamentos de Qu mica, pp 133135, M exico: Editorial Continental Brierley, CL (1995). Bacterial oxidation: revolutionizing the gold industry. Australian Gold Conference, Kalgoorlie, Australia C ordova, R, G omez, H, Real, SG, Schrebler, R and Vilche, JR (1997). J Electrochem Soc 144: 26282636 Escobar, B, Huenupi, E and Wiertz, JV (1997). Biotechnol Lett 19: 719722 Flashka, HA, Barnard, AJ and Sturrock, PE (1969). Quantitative Analytical Chemistry, Vol. 1. pp. 195197, New York: Barnes and Noble Herrera, L, Ruiz, P, Aguill on, JC and Fehrmann, A (1989). J Chem Technol Biotechnol 44: 171181 Nagpal, S, Dahlstrom, D and Oolman, T (1993). Biotechnol Bioeng 41: 459464 Silverman, MP and Lundgren, DG (1959). J Bacteriol 77: 642647 Sobotka, M, Prokop, A, Dunn, IJ and Einsele, A (1982). Review of methods for the measurement of oxygen transfer in microbial systems. In : Annual Report on Fermentation Processes, Vol. 5. GT Tsao, ed pp 127210, New York: Academic Press

The inhibition of T. ferrooxidans growth and leaching activity by high carbon dioxide partial pressures has been noted previously (Nagpal et al., 1993). The effect of CO2 enrichment was also reected in the degree of extraction obtained in each run (Table 3). Again, 4% carbon dioxide gave maximal Fe, Cu and As extractions. At this condition, extractions increased by factors of 3.3 for Fe, 1.2 for Cu, and 1.3 for As. Although extractions were far from complete, they caused an increase in gold recovery by cyanidation of over 100% (results not shown). The high gures reported for iron in Tables 2 and 3 imply a preferential biooxidation of pyrite over enargite, which is a very refractory mineral with a slow dissolution rate (Escobar et al., 1997). According to the results obtained by C ordova et al. (1997) working in the electrochemical oxidation of this same concentrate, a much higher dissolution rate would be expected if the Eh values could be raised to 600 mV or higher. The differences in the optimal CO2 concentrations reported by different authors are due to differences in mineral composition, microbial strains and operating conditions. In this respect, the mass transfer capability of the equipment is important. The bioreactor used in this work showed a kLa for oxygen of 60.7 h1, which gives a calculated value for the kLa of CO2 of 51.6 h1. The measured value for this coefcient was 47.8 h1. It is considered that the small difference between them is acceptable and in the range of experimental error.

Received: 10 December Revisions requested: 19 December Revisions received: 22 January Accepted: 23 January

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