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Thesis (Ph.D. (Biochemistry)) --University of Limpopo, 2010 / A heavy-metal resistant bacterium (GM 16) was isolated from a South African antimony mine, and the non-viable cells of the isolate were used to investigate its biosorption capacity for Cd(II) and Cu(II) from aqueous solution in a batch process. The biosorption of both metals were found to be influenced by factors such as pH of the metal solution, initial metal ion and biomass concentrations, rate of agitation, presence of other metal ions, contact time of the metal solution with the biomass and temperature. The initial biosorption of both Cd(II) and Cu(II) was rapid and equilibrium was reached within 1 hour of biomass contact with the metal solutions. The sorption of both metal ions was higher in weak acid than in strong acid conditions and the optimum pH values for Cd(II) and Cu(II) biosorption were 7 and 6, respectively. The presence of the other metal ions in the metal adsorption media influenced the biosorption of both Cd(II) and Cu(II). Mg2+ ions decreased the uptake of Cu(II) and Cd(II) by 4.7 and 6.5 %, respectively. Whereas K+, Na+ and Ca2+ ions increased the uptake of Cd(II) by 12.3, 8.7, and 3.2 %, respectively, they slightly decreased the sorption Cu(II) (2-6.4 %). Increases in initial metal ion (40-120 mg L-1) and biomass (0.8-4.8 g L-1) concentrations enhanced the sorption of Cd(II) and Cu(II) by GM16 biomass. When the biomass concentration was increased from 0.8 to 4.8 g L-1, the biosorption capacity of Cd(II) increased from 5.5 to 14.5 mg g-1 while that of Cu(II) increased from 2.8 to 14.7 mg g-1 at optimum pHs and a temperature of 40 °C. Maximum adsorption of both metals occurred at an agitation rate of 100 rpm. In addition, increase in initial metal ion concentration from 40-120 mg L-1 increased the initial adsorption rates (h) and the equilibrium metal sorption capacity (qe) of the GM 16 biomass from 6.07 to 16.51 mg g-1 for Cu(II) and 8.9 to 17.9 mg g g-1 for Cd(II). Adsorption equilibrium data for both metal ions fitted well to the Langmuir adsorption model with high correlation coefficients (r2 > 0.90) but the data for Cu(II) could also be described by the Freundlich adsorption model. Increase in temperature from 25-40 °C only caused marginal increases in maximum metal sorption capacities (qmax). The results on kinetic analysis showed that the biosorption processes of Cd(II) and Cu(II) ions by the non-viable GM 16 cells followed pseudo-second order kinetic model betterthan the pseudo-first order model, although the calculated metal sorption capacities obtained with the model were overestimated. The calculated thermodynamic parameters showed that the biosorption of Cd(II) and Cu(II) ions was feasible, spontaneous and slightly endothermic for Cd(II) but slightly exothermic for Cu(II) under examined conditions. Based on 16S ribosomal DNA sequencing, the bacterial isolate (GM 16) was identified as a Bacillus sp. and is closely related to Bacillus thuringiensis and Bacillus cereus strains. The biosorption capacity of the non-viable GM 16 biomass was higher than the biosorption capacity reported for the viable GM 16 cells, 65 % of Cd(II) was removed by non-viable biomass whereas 48 % was removed by the viable biomass. For the biosorption of Cu(II), the % metal ion adsorbed for the non-viable GM 16 cells was slightly higher than the % adsorbed for the viable cells although not statistically significant. Only 67 % of Cu(II) was removed by the non-viable cells whereas 65 % was removed by the viable cells. / National Research Foundation

A mina do Sossego, localizada em Canaã dos Carajás - Pará, principal mina de cobre do país, atualmente possui uma lagoa de rejeitos com baixas concentrações de cobre. Este é um cenário de impacto ambiental, que pode ser diminuído através de processos de biorremediação, usando a microbiota local. Neste contexto, este trabalho avalia a capacidade de biossorção de cobre (Cu++) por linhagens bacterianas isoladas de amostras ambientais da mina, bem como as melhores condições para o processo. Dentre as linhagens isoladas de amostras ambientais (água e sedimento), duas delas foram exploradas nesse trabalho quanto à resistência e capacidade de biossorção de cobre, uma vez que mostraramse representativas da microbiota e tolerantes à concentrações de até 400 mg.L-1 de Cu++. Estas linhagens foram identificadas como Enterobacter cloacae e Rhodococcus erythropolis por espectrometria de massas (MALDI-TOF) e sequenciamento do gene 16S rRNA, apresentando capacidades máximas de biossorção de Cu++ de aproximadamente 122,0 mg.g- 1 e 370,0 mg.g-1 para R. erythropolis e E. cloacae, respectivamente, nas condições estudadas. Os parâmetros cinéticos de isotermas de biossorção foram obtidos através do modelo de Langmuir. As melhores condições para o processo de biossorção de Cu++ encontradas foram de 28°C, 180 rpm e 0,1g.L-1 de biomassa para, pH de 5 e 6, respectivamente para R.erythropolis e E.cloacae. Dados de bioacumulação também foram observados, sugerindo um mecanismo de efluxo celular para ambos os isolados, apresentando dados de captação máxima de Cu++ de 210 mg.g-1, para R. erythropolis e 20 mg.g-1, para E. cloacae. Também foram avaliadas diferentes condições de cultivo para maior obtenção de biomassa bacteriana, conseguindo um aumento de rendimento celular de até 7 vezes para R. erythropolis e 3 vezes para E.cloacae. / The Sossego mine, located in Canaã dos Carajás - Pará, currently has a tailings pond with low concentration of copper. This is an environmental impact scenario, which can be decreased by bioremediation processes using the local microbiota. In this context, this work evaluates the copper biosorption capacity (Cu++) from bacterial strains isolated from environmental samples of the mine, as well as the best conditions for the process. Among the isolated strains of environmental samples (water and sediment), two of them were explored in this work regarding the resistance and capacity of copper biosorption, because they were abundant and tolerant to concentrations up to 400 mg.L-1 Cu++. These strains were identified as Enterobacter cloacae and Rhodococcus erythropolis by MALDI-TOF and 16S gene sequencing, and showed maximum copper biosorption capacities of approximately 122.0 mg.g-1 and 370.0 mg.g-1 for R. erythropolis and E. cloacae, respectively, in the better conditions studied, according to kinetic parameters of biosorption isotherms, through the Langmuir model. The best conditions for the copper biosorption process were 28 °C, 180 rpm and 0.1g.L-1 of biomass for both, and pH of 5 and 6, respectively, for R .erythropolis and E. cloacae. Bioaccumulation data were also observed, suggesting a mechanism of cellular efflux for both bacteria and presenting data of maximum copper uptake of 210 mg.g-1, for R. erythropolis and 20 mg.g-1, for E. cloacae. Different cultivation conditions were also evaluated to obtain a higher bacterial biomass, achieving a cellular yield increase of up to 7 times for R. erythropolis and 3 times for E. cloacae.

A mina do Sossego, localizada em Canaã dos Carajás - Pará, principal mina de cobre do país, atualmente possui uma lagoa de rejeitos com baixas concentrações de cobre. Este é um cenário de impacto ambiental, que pode ser diminuído através de processos de biorremediação, usando a microbiota local. Neste contexto, este trabalho avalia a capacidade de biossorção de cobre (Cu++) por linhagens bacterianas isoladas de amostras ambientais da mina, bem como as melhores condições para o processo. Dentre as linhagens isoladas de amostras ambientais (água e sedimento), duas delas foram exploradas nesse trabalho quanto à resistência e capacidade de biossorção de cobre, uma vez que mostraramse representativas da microbiota e tolerantes à concentrações de até 400 mg.L-1 de Cu++. Estas linhagens foram identificadas como Enterobacter cloacae e Rhodococcus erythropolis por espectrometria de massas (MALDI-TOF) e sequenciamento do gene 16S rRNA, apresentando capacidades máximas de biossorção de Cu++ de aproximadamente 122,0 mg.g- 1 e 370,0 mg.g-1 para R. erythropolis e E. cloacae, respectivamente, nas condições estudadas. Os parâmetros cinéticos de isotermas de biossorção foram obtidos através do modelo de Langmuir. As melhores condições para o processo de biossorção de Cu++ encontradas foram de 28°C, 180 rpm e 0,1g.L-1 de biomassa para, pH de 5 e 6, respectivamente para R.erythropolis e E.cloacae. Dados de bioacumulação também foram observados, sugerindo um mecanismo de efluxo celular para ambos os isolados, apresentando dados de captação máxima de Cu++ de 210 mg.g-1, para R. erythropolis e 20 mg.g-1, para E. cloacae. Também foram avaliadas diferentes condições de cultivo para maior obtenção de biomassa bacteriana, conseguindo um aumento de rendimento celular de até 7 vezes para R. erythropolis e 3 vezes para E.cloacae. / The Sossego mine, located in Canaã dos Carajás - Pará, currently has a tailings pond with low concentration of copper. This is an environmental impact scenario, which can be decreased by bioremediation processes using the local microbiota. In this context, this work evaluates the copper biosorption capacity (Cu++) from bacterial strains isolated from environmental samples of the mine, as well as the best conditions for the process. Among the isolated strains of environmental samples (water and sediment), two of them were explored in this work regarding the resistance and capacity of copper biosorption, because they were abundant and tolerant to concentrations up to 400 mg.L-1 Cu++. These strains were identified as Enterobacter cloacae and Rhodococcus erythropolis by MALDI-TOF and 16S gene sequencing, and showed maximum copper biosorption capacities of approximately 122.0 mg.g-1 and 370.0 mg.g-1 for R. erythropolis and E. cloacae, respectively, in the better conditions studied, according to kinetic parameters of biosorption isotherms, through the Langmuir model. The best conditions for the copper biosorption process were 28 °C, 180 rpm and 0.1g.L-1 of biomass for both, and pH of 5 and 6, respectively, for R .erythropolis and E. cloacae. Bioaccumulation data were also observed, suggesting a mechanism of cellular efflux for both bacteria and presenting data of maximum copper uptake of 210 mg.g-1, for R. erythropolis and 20 mg.g-1, for E. cloacae. Different cultivation conditions were also evaluated to obtain a higher bacterial biomass, achieving a cellular yield increase of up to 7 times for R. erythropolis and 3 times for E. cloacae.