Direct electron uptake from a cathode using the inward Mtr pathway in Escherichia coli

Research on the biocathode-based bioelectrochemical system (BES) has attracted extensive attention because of its ability to increase the electricity-driven production of high-value fuels or chemicals by relying on microbial cells as catalysts. An extracellular electron transfer (EET) that makes electrical connections to microorganisms plays a key role in the BES. Compared with the better understanding of the EET-to-anode connection, the understanding of the mechanism and elements involved in inward EET from cathodes to microbes remains limited. Additionally, the low capability of the EET limits its applications in BESs for producing chemicals. Here, we introduced the Mtr pathway into Escherichia coli cells by expressing ccmABCDEFGH from E. coli and mtrABC from Shewanella oneidensis. Through selection by electrochemical pressure, the evolved E. coli could use electricity to increase the production of succinate in direct BES and enhance the electroactivity. In addition, in studying the mechanism of inward EET, menaquinone was found to be one of the key components of inward EET, and it is essential for the fumarate reduction reaction. Lastly, the intracellular NADH and ATP levels showed that there were differences in the energy conservation coupling between the electron transfer routes of the inward Mtr pathway and the electron mediator.