Inhibition sensitivity of in vitro firefly bioluminescence quantum yields to Zn2+ and Cd2+ concentrations in aqueous solutions
Abstract
Abstract To elucidate the inhibition effects of Zn2+ and Cd2+ on the luciferin–luciferase reaction, we performed quantitative measurements of quantum yields and spectral shapes for in vitro firefly bioluminescence in aqueous solutions containing ZnSO4, ZnCl2, CdSO4, and CdCl2 at different concentrations. Particular care was taken toward the equilibrium between metal ions and enzyme proteins, anion difference, solubility, and uncertainty evaluation. The bioluminescence quantum yields decreased almost linearly to the concentration of Zn2+ and Cd2+ below 0.25 mM. No obvious difference was found between the chloride and sulfate anion solutions. We defined inhibition sensitivity as the decrease in relative quantum yield versus the concentration of metal ions, and they were determined to be 1.48 ± 0.13 and 1.13 ± 0.16/mM for Zn2+ and Cd2+, respectively. We estimated the detection limit of inhibition effects as the concentration of metal ions that decrease relative quantum yields by 10%, which were 0.07 mM (4 ppm) and 0.09 mM (10 ppm) for Zn2+ and Cd2+, respectively. The shape of the bioluminescence spectra changed sensitively with the increase in Zn2+ concentrations. The bioluminescence peak energy for 0.10‐mM Zn2+ was ~2.2 eV, while that for 0.25‐mM Zn2+ was ~2.0 eV. The shape of the spectra changed less sensitively with the increase in Cd2+concentrations, and the peak energy was at ~2.2 eV for Cd2+ concentrations of 0.10 and 0.25 mM.
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