Effect of complexing ligands on the surface adsorption, internalization, and bioresponse of copper and cadmium in a soil bacterium, Pseudomonas putida
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Environmental quality criteria for metals toxic to soil and water organisms, using the Free Ion Activity Model or the Biotic Ligand Model, are based on the concept that the major form of the metal available to the organism is the free metal ion, yet various metal complexes are bioavailable to a variety of soil and water organisms. We test here whether neutral copper or cadmium sulfates, negatively-charged copper or cadmium citrates and positively-charged copper acetate and cadmium chloride are bioavailable to a soil bacterium, Pseudomonas putida. Adsorption onto the cell surface and uptake into the periplasm and cytoplasm of this Gram-negative root colonizing bacterium was studied by adding a single concentration of Cu or Cd and varying the concentration of the ligands to complex 10 to 100% of the metal. Metal association from the complexes on and within the cell was defined using selective extraction procedures and compared with free ion controls using the Langmuir isotherm. Cellular responses also were assessed using a P. putida biosensor. Both uptake and bioresponse methodologies showed that P. putida was sensitive to the metal complexes. In particular, the bioresponse to Cu and Cd supplied as a citrate complex occurred with activities of free metal ions two orders of magnitude lower than for the control. We concluded that the tested metal complexes for Cu and Cd are taken up into the cell, where they trigger a bioresponse. We also discuss the implications of these findings on interactions between soil and water organisms and nanoparticles that release metal ions.