Equilibrium, kinetic, and thermodynamic studies on the biosorption of lead by human metallothionein gene-cloned bacteria as a novel biosorbent.

Heavy metals are the main pollutants in water and are an important global problem that threatens human health and ecosystems. In recent years, there has been an increasing interest in the use of genetically modified bacteria as an eco-friendly method to solve heavy metal pollution problems. The goal of this study was to generate genetically modified Escherichia coli expressing human metallothioneins (hMT2A and hMT3) and to determine their tolerance, bioaccumulation, and biosorption capacity to lead (Pb2+ ). Recombinant MT2A and MT3 strains expressing MT were successfully generated. Minimum inhibition concentrations (MIC) of Pb for MT2A and MT3 were found to be 1750 and 2000 mg L-1 , respectively. Pb2+ resistance and bioaccumulation capacity of MT3 were higher than MT2A. Therefore, only MT3 biosorbent was used in Pb2+ biosorption, and its efficiency was examined by performing experiments in a batch system. Pb2+ biosorption by MT3 was evaluated in terms of isotherms, kinetics, and thermodynamics. The results showed that Pb biosorption fits to the Langmuir isotherm model and the pseudo-first-order kinetic model, and the reaction is exothermic. The maximum Pb2+ capacity of the biosorbent was 50 mg Pb2+ g-1 . The potential of MT3 in Pb biosorption was characterized by Fourier-transform infrared spectroscopy (FT-IR), scanning electron microscopy (SEM), and scanning transmission electron microscopy (STEM) analyses. The desorption study showed that the sorbent had up to 74% recovery and could be effectively used four times. These findings imply that this biosorbent can be applied as a promising, precise, and effective means of removing Pb2+ from contaminated waters. PRACTITIONER POINTS: In this study, the tolerance levels, bioaccumulation, and biosorption capacities of Pb in aqueous solutions were determined for the first time in recombinant MT2A and MT3 strains in which human MT2A and MT3 genes were cloned. The biosorbent of MT3, which was determined to be more effective in Pb bioaccumulation, was synthesized and used in Pb biosorption. The Pb biosorption mechanism of MT3 biosorbent was identified using isotherm modeling, kinetic modeling, and thermodynamic studies. The maximum Pb removal percentage capacity of the biosorbent was 90%, whereas the maximum biosorption capacity was up to 50 mg Pb2+ g-1 . These results indicated that MT3 biosorbent has a higher Pb biosorption capacity than existing recombinant biosorbents. MT3 biosorbent can be used as a promising and effective biosorbent for removing Pb from wastewater.

Production of methionine γ- lyase in recombinant Citrobacter freundii bearing the hemoglobin gene.

The production of antileukemic enzyme methionine γ-lyase (MGL) in distinctly related bacteria, Citrobacter freundii and in their recombinants expressing the Vitresocilla hemoglobin (VHb) has been studied. This study concerns the potential of Citrobacter freundii expressing the Vitreoscilla hemoglobin gene (vgb) for the methionine γ- liyase production. Methionine γ- liyase production by Citrobacter freundii and its vgb(-) and vgb(+) bearing recombinant strain was studied in shake-flasks under 200 rpm agitation, culture medium and 30 °C in a time-course manner. The vgb(+) and especially the carbon type had a dramatic effect on methionine γ- liyase production. The vgb(+) strain of C. freundii had about 2-fold and 3.1-fold higher levels of MGL than the host and vgb(-) strain, respectively.

Genetic engineering of Enterobacter aerogenes with the Vitreoscilla hemoglobin gene: cell growth, survival, and antioxidant enzyme status under oxidative stress.

Hemoglobins in unicellular organisms, like the one here in the bacterium Vitreoscilla, have greater chemical reactivity than their homologues in multicellular organisms. They can catalyze redox reactions and may protect cells against oxidative stress. The ability of Vitreoscilla hemoglobin to complement deficiencies of terminal cytochrome oxidases in Escherichia coli also suggests that this hemoglobin can receive electrons during respiration. In this study, a recombinant strain of Enterobacter aerogenes engineered to produce the Vitreoscilla Hb was investigated with regard to its susceptibility to oxidative stress. The culture response to oxidative stress produced by exogenously applied hydrogen peroxide was characterized in terms of cell growth, survival and the activities of two key antioxidant enzymes (catalase and superoxide dismutase). The influence of the physiological state of the cells and different media upon these culture dynamics was determined. Results showed that the hemoglobin-expressing strain is quite distinct in terms of growth/survival properties and activity of antioxidant enzymes from that of non-hemoglobin counterparts.