RESUMO
Bacterial strains from karst landform soil were enriched via chemostat culture in the presence of sodium bicarbonate. Two chemolithotrophic strains were isolated and identified as Serratia marcescens Wy064 and Bacillus sp. Wy065. Both strains could grow using sodium bicarbonate as the sole carbon source. Furthermore, the supplement of the medium with three electron donors (Na2S, NaNO2, and Na2S2O3) improved the growth of both strains. The activities of carbonic anhydrase (CA) and ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) could be detected in the crude enzyme of strain Wy064, implying that the strain Wy064 might employ Calvin cycle to fix CO2. S. marcescens genome mining revealed four potential CA genes designated CA1-CA4. The proteins encoded by genes CA1-3 were cloned and expressed in Escherichia coli. The purified recombinant enzymes of CA1 and CA3 exhibited CO2 hydration activities, whereas enzyme CA2 was expressed in inclusion bodies. A CO2 hydration assay demonstrated that the specific activity of CA3 was significantly higher than that of CA1. The maximum CO2 hydration activities for CA1 and CA3 were observed at pH 7.5 and 40 °C. The activities of CA1 and CA3 were significantly enhanced by several metal ions, especially Zn2+, which resulted in 21.1-fold and 26.1-fold increases of CO2 hydration activities, respectively. The apparent K m and V max for CO2 as substrate were 27 mM and 179 WAU/mg for CA1, and 14 mM and 247 WAU/mg for CA3, respectively. Structure modeling combined with sequence analysis indicated that CA1 and CA3 should belong to the Type II ß-CA.
RESUMO
The mechanism of uranium transformation from U(VI) into nano-uramphite by two indigenous Bacillus thuringiensis strains was investigated in the present work. Our data showed that the bacteria isolated from uranium mine possessed highly accumulation ability to U(VI), and the maximum accumulation capacity was around 400 mg U/g biomass (dry weight). X-ray powder diffraction (XRD), scanning electron microscopy (SEM), transmission electron microscopy (TEM) and Fourier transform infrared spectroscopy (FT-IR) analyzes indicated that the U(VI) was adsorbed on the bacterial surface firstly through coordinating with phosphate, CH2 and amide groups, and then needle-like amorphous uranium compounds were formed. With the extension of time, the extracellular crystalline substances were disappeared, but some particles were appeared in the intracellular region, and these particles were characterized as tetragonal-uramphite. Moreover, the disrupted experiment indicated that the cell-free extracts had better uranium-immobilization ability than cell debris. Our findings provided the understanding of the uranium transformation process from amorphous uranium to crystalline uramphite, which would be useful in the regulation of uranium immobilization process.
