Sample injections at different times. It should be noted that y-scale is higher for chromatogram B. Under these conditions, CA activity was linear for at least 1 min (Fig. S4). Reverse reaction in turn, produced less reliable values by this method (not shown). (TIF) Figure S4 Activity of CA in the cytosolicfraction of M. acetivorans in the absence ( ) or presence of 0.01 (N), 0.1 (m), 1 (.) or 10 mM total CdCl2 (X). A representative data with heated cytosolic fraction in presence of 0.1 mM CdCl2 is also shown ( ). (TIF) Figure S5 Activation of methanogenesis by cadmium. Cultures on acetate were GW 0742 custom synthesis purged by passing N2 for 5 min. Then, samples of the head space were withdrawn from the cultures at 0 and 5, 10, 20, 30 and 60 min of incubation with 0 (filled symbols) or 10 mM CdCl2 (open symbols) for GC analysis. These experiments were started with the addition of 20 mM acetate. Values are the mean 6 SD of 3 independent cell preparations. P,0.05 using the Student’s t-test for a vs control (without cadmium). 18325633 (TIF) Figure S6 Intracellular cadmium clusters in M. acetivorans. HAADF-STEM projection images of methanol-grown cells cultured in methanol in the absence (A) or in the presence of 100 mM CdCl2 for 5 days (B). The image in B revealed cadmium grains along the cell (white spots). (TIF) Text S1 Methods and Results.Concluding remarksDespite the very low concentration calculated of free Cd2+, this non-essential heavy metal was able to activate a biological process, i.e., methanogenesis in M. acetivorans, due in part to a direct activation of acetoclastic pathway enzymes. M. acetivorans removed and accumulated cadmium; hence, M. acetivorans may become a suitable model for studying the effect of heavy metals on marine methanogens and its mechanisms of heavy metal resistance in the Archaea domain. Moreover, further optimization of the enhanced methane production by cadmium, and other heavy metals, may place this process in the biotechnological leading frontier for generation of biogas.(DOCX)Supporting InformationFigure S1 Activity of phosphotransacetylase from M.AcknowledgmentsAuthors thank Dr. Karla Carvajal-Aguilera for assistance in the statistical analysis. The authors also thank Patricia Bizarro Nevares (Academic technician) and Armando Zepeda Rodriguez (coordinator of the Laboratory of electron microscopy) from the Department of Cellular and Tissue Biology, University of Mexico for the preparation of the cell samples for the ultrastructure analysis, and to Physicist Roberto Hernandez Reyes ?(Academic technician) for the electron Eliglustat microscopy X-ray analysis at the Physics Institute, University of Mexico.acetivorans. An aliquot of the cytosolic fraction was incubated with the substrates acetyl-Pi and CoA in the absence (( ) or presence of 0.1 (N), 1 (m) or 10 (.) mM total CdCl2. In the absence of protein the CoA concentration remained constant ( ). (TIF)Figure S2 CODH/AcCoAs complex activity from M. acetivorans; representative traces of the activity by adding cytosolic fraction (containing the enzyme complex, ferredoxin and THMPT) and 80 mM AcCoA. Representative trace with: 125 ( ), 62 (x) and 25 ( ) mg of cytosolic fraction without cadmium in the absence or presence of 0.01 (N), 0.1 (m), 1 (.) and 10 mM total CdCl2 (X). (TIF)Author ContributionsConceived and designed the experiments: RJC RMS. Performed the experiments: ELS MGSM VHJ RGC. Analyzed the data: RJC. Contributed reagents/materials/analysis tools: RJC. Wrote the paper: RJC RMS.
Cancer st.Sample injections at different times. It should be noted that y-scale is higher for chromatogram B. Under these conditions, CA activity was linear for at least 1 min (Fig. S4). Reverse reaction in turn, produced less reliable values by this method (not shown). (TIF) Figure S4 Activity of CA in the cytosolicfraction of M. acetivorans in the absence ( ) or presence of 0.01 (N), 0.1 (m), 1 (.) or 10 mM total CdCl2 (X). A representative data with heated cytosolic fraction in presence of 0.1 mM CdCl2 is also shown ( ). (TIF) Figure S5 Activation of methanogenesis by cadmium. Cultures on acetate were purged by passing N2 for 5 min. Then, samples of the head space were withdrawn from the cultures at 0 and 5, 10, 20, 30 and 60 min of incubation with 0 (filled symbols) or 10 mM CdCl2 (open symbols) for GC analysis. These experiments were started with the addition of 20 mM acetate. Values are the mean 6 SD of 3 independent cell preparations. P,0.05 using the Student’s t-test for a vs control (without cadmium). 18325633 (TIF) Figure S6 Intracellular cadmium clusters in M. acetivorans. HAADF-STEM projection images of methanol-grown cells cultured in methanol in the absence (A) or in the presence of 100 mM CdCl2 for 5 days (B). The image in B revealed cadmium grains along the cell (white spots). (TIF) Text S1 Methods and Results.Concluding remarksDespite the very low concentration calculated of free Cd2+, this non-essential heavy metal was able to activate a biological process, i.e., methanogenesis in M. acetivorans, due in part to a direct activation of acetoclastic pathway enzymes. M. acetivorans removed and accumulated cadmium; hence, M. acetivorans may become a suitable model for studying the effect of heavy metals on marine methanogens and its mechanisms of heavy metal resistance in the Archaea domain. Moreover, further optimization of the enhanced methane production by cadmium, and other heavy metals, may place this process in the biotechnological leading frontier for generation of biogas.(DOCX)Supporting InformationFigure S1 Activity of phosphotransacetylase from M.AcknowledgmentsAuthors thank Dr. Karla Carvajal-Aguilera for assistance in the statistical analysis. The authors also thank Patricia Bizarro Nevares (Academic technician) and Armando Zepeda Rodriguez (coordinator of the Laboratory of electron microscopy) from the Department of Cellular and Tissue Biology, University of Mexico for the preparation of the cell samples for the ultrastructure analysis, and to Physicist Roberto Hernandez Reyes ?(Academic technician) for the electron microscopy X-ray analysis at the Physics Institute, University of Mexico.acetivorans. An aliquot of the cytosolic fraction was incubated with the substrates acetyl-Pi and CoA in the absence (( ) or presence of 0.1 (N), 1 (m) or 10 (.) mM total CdCl2. In the absence of protein the CoA concentration remained constant ( ). (TIF)Figure S2 CODH/AcCoAs complex activity from M. acetivorans; representative traces of the activity by adding cytosolic fraction (containing the enzyme complex, ferredoxin and THMPT) and 80 mM AcCoA. Representative trace with: 125 ( ), 62 (x) and 25 ( ) mg of cytosolic fraction without cadmium in the absence or presence of 0.01 (N), 0.1 (m), 1 (.) and 10 mM total CdCl2 (X). (TIF)Author ContributionsConceived and designed the experiments: RJC RMS. Performed the experiments: ELS MGSM VHJ RGC. Analyzed the data: RJC. Contributed reagents/materials/analysis tools: RJC. Wrote the paper: RJC RMS.
Cancer st.
