Abstract
Microorganisms contribute to the formation of secondary gold (Au) deposits through enzymatic reduction of Au(III) to Au(0). However, the enzyme that catalyzes the reduction of Au(III) remains enigmatic. Here, we identified and characterized a previously unknown Au reductase (GolR) in the cytoplasm of Erwinia sp. IMH. The expression of golR was strongly up-regulated in response to increasing Au(III) concentrations and exposure time. Mutant with in-frame deletion of golR was incapable of reducing Au(III), and the capability was rescued by reintroducing wild-type golR into the mutant strain. The Au(III) reduction was determined to occur in the cytoplasmic space by comparing the TEM images of the wild-type, mutant, and complemented strains. In vitro assays of the purified GolR protein confirmed its ability to reduce Au(III) to Au nanoparticles. Molecular dynamic simulations demonstrated that the hydrophobic cavity of GolR may selectively bind AuCl(2)(OH)(2) (-), the predominant auric chloride species at neutral pH. Density functional theory calculations revealed that AuCl(2)(OH)(2) (-) may be coordinated at the Fe-containing active site of GolR and is probably reduced via three consecutive proton-coupled electron transfer processes. The new class of reductase, GolR, opens the chapter for the mechanistic understanding of Au(III) bioreduction.