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The mechanisms by which  Geobacter sulfurreducens  transfers electrons through relatively thick (>50 µm) biofilms to electrodes acting as a sole electron acceptor were investigated. Biofilms of  Geobacter sulfurreducens  were grown either in flow-through systems with graphite anodes as the electron acceptor or on the same graphite surface, but with fumarate as the sole electron acceptor. Fumarate-grown biofilms were not immediately capable of significant current production, suggesting substantial physiological differences from current-producing biofilms. Microarray analysis revealed 13 genes in current-harvesting biofilms that had significantly higher transcript levels. The greatest increases were for  pilA , the gene immediately downstream of  pilA , and the genes for two outer  c -type membrane cytochromes, OmcB and OmcZ. Down-regulated genes included the genes for the outer-membrane  c -type cytochromes, OmcS and OmcT. Results of quantitative RT-PCR of gene transcript levels during biofilm growth were consistent with microarray results. OmcZ and the outer-surface  c -type cytochrome, OmcE, were more abundant and OmcS was less abundant in current-harvesting cells. Strains in which  pilA , the gene immediately downstream from  pilA ,  omcB ,  omcS ,  omcE , or  omcZ  was deleted demonstrated that only deletion of  pilA  or  omcZ  severely inhibited current production and biofilm formation in current-harvesting mode. In contrast, these gene deletions had no impact on biofilm formation on graphite surfaces when fumarate served as the electron acceptor. These results suggest that biofilms grown harvesting current are specifically poised for electron transfer to electrodes and that, in addition to pili, OmcZ is a key component in electron transfer through differentiated  G. sulfurreducens  biofilms to electrodes.

Anode Biofilm Transcriptomics Reveals Outer Surface Components Essential for High Density Current Production in Geobacter sulfurreducens Fuel Cells