The effect of Zn 2+ on glucose 6-phosphate dehydrogenase activity from Bufo arenarum toad ovary and alfalfa plants

Abstract

The effect of Zn 2+ on glucose 6-phosphate dehydrogenase (G6PD) activity was monitored in samples from Bufo arenarum toad ovary and alfalfa plants, in the search for a possible new bioindicator able to detect levels of exposure through contaminated soils, and also to elucidate possible similarities between the enzyme from animal and plant tissues. The in vivo effect was evaluated after exposure of the toads to the metal in Ringer solution during 30 days and after 10 days of treatment in 6 weeks old plants, cultured under laboratory conditions. In vitro effects were measured in different extracts from control samples and partially purified enzyme from ovarian tissue as well as in different extracts from control alfalfa plants, by addition of the metal to the reaction mixture containing the enzyme. G6PD from toad ovary was noncompetitively inhibited by zinc both in vivo and in vitro, under all the experimental conditions studied. A kinetic analysis of the enzyme activity showed that the Michaelis–Menten constant ( K m) was not modified, while maximal velocity ( V max) decreased as the consequence of treatment. It was not possible to obtain a dose–response curve for the effects of Zn 2+ on G6PD from alfalfa whole plants, measured in vivo or in vitro. Only leaf extracts evidenced a possible relationship between treatment with the metal and G6PD activity alteration. The results agree with a possible role for G6PD as a biomarker of effect and exposure to Zn 2+ in B. arenarum ovarian tissue but not in alfalfa plants.