Geobacter cytochrome OmcZs binds riboflavin: implications for extracellular electron transfer

Geobacter sulfurreducens is an important model organism for understanding extracellular electron transfer (EET), i.e. transfer of electrons from the cell's interior (quinone pool) to an extracellular substrate. This exoelectrogenic functionality can be exploited in bioelectrochemical applications. Nonetheless, key questions remain regarding the mechanisms of this functionality. G. sulfurreducens has been hypothesized to employ both multi-heme cytochromes and soluble, small molecule redox shuttles, as the final, redox-active species in EET. However, interactions between flavin redox shuttles and outer membrane, redox proteins in Geobacter have not been demonstrated. Herein, the heterologous expression and purification from E. coli of a soluble form of the multi-heme cytochrome OmcZs from G. sulfurreducens is reported. UV–vis absorption assays show that riboflavin can be reduced by OmcZs with concomitant oxidation of the protein. Fluorescence assays show that oxidized OmcZs and riboflavin interact with a binding constant of 34 μM. Furthermore, expression of OmcZs in E. coli enables EET in the host, and the current produced by these E. coli in a bioelectrochemical cell increases when riboflavin is introduced. These results support the hypothesis that OmcZs functions in EET by transiently binding riboflavin, which shuttles electrons from the outer membrane to the extracellular substrate.