Cytochrome c Reductase is a Key Enzyme Involved in the Extracellular Electron Transfer Pathway towards Transition Metal Complexes in Pseudomonas Putida

Mediator-based extracellular electron transfer (EET) pathways can balance the redox metabolism of microbes. However, such electro-biosynthesis processes are constrained by the unknown underlying EET mechanisms. In this paper, we studied Pseudomonas putida to systematically investigate its EET pathway to transition metal complexes (i.e. [Fe(CN) 6 ] 3-/4- and [Co(bpy) 3 ] 3+/2+ ) under anaerobic conditions. Comparative proteomics showed the aerobic respiratory components were upregulated in a bioelectrochemical system without oxygen, suggesting their potential contribution to EET. Further tests found inhibiting cytochrome c oxidase activity by NaN 3 and NADH dehydrogenase by rotenone did not significantly change the current output. However, the EET pathway was completely blocked, while cytochrome c reductase activity was inhibited by antimycin A. Although it cannot be excluded that cytochrome c and the periplasmic subunit of cytochrome c oxidase donate electrons to the transition metal complexes, these results strongly demonstrate that cytochrome c reductase is a key complex for the EET pathway.