Abstract
A glutathione reductase (GSHR)-like enzyme in Pseudomonas moraviensis stanleyae was previously implicated as underlying the bacterium's remarkable SeO3 2- tolerance. Herein, this enzyme is sequenced, recombinantly expressed, and fully characterized. The enzyme is highly adapted for selenodiglutathione substrates (K m = 336 μM) compared to oxidized glutathione (K m = 8.22 mM). The recombinant expression of this enzyme in the laboratory strains of Escherichia coli conveys a 10-fold increase in IC90 for SeO3 2-. Moreover, selenium nanoparticles are observed when the enzyme is overexpressed in the cells exposed to SeO3 2-, but not in the corresponding no-enzyme controls. The analyses of the structural homology models of the enzyme reveal changes in the parts of the enzyme associated with product release, which may underlie the Se substrate specialization. Combined, the observations of adaptation to Se reduction over oxidized glutathione reduction as well as the portability of this nanoparticle-mediated SeO3 2- tolerance into other cell lines suggest that the P. moraviensis GSHR may be better described as a GSHR-like metalloid reductase.