EXPRESSION OF THE NEUROSPORA-CRASSA METALLOTHIONEIN GENE IN ESCHERICHIA-COLI AND ITS EFFECT ON HEAVY-METAL UPTAKE

The gene coding for the Neurospora crassa metallothionein protein was chemically synthesized and cloned into the fusion expression vectors pMal-c and pMal-p. Cell-fractionation experiments demonstrated the proper localization of the pMal-c- and pMal-p- expressed proteins to the cytosol and periplasm of the bacteria respectively. Control bacteria as well as the recombinant bacteria producing the metallothionein protein were incubated with solutions of Cd-109 at concentrations of 0.2 mu M, 1 mu M, and 10 mu M. The recombinant bacteria were able to accumulate significantly more Cd-109 than control bacteria at al concentrations tested. Cadmium accumulation was rapid and highly selective. Maximum uptake was achieved at a pH of 7.0, with lower accumulation at lower or higher pH values. The pH-dependent uptake of cadmium by the recombinant bacteria was exploited to strip off the bound cadmium from the recombinant bacteria and to regenerate most of the cadmium-binding sites. These observations suggest the potential for using a metallothionein-based biosorbent for certain heavy-metal removal applications.