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Mohammed Mouhib, Melania Reggente, Lin Li, Nils Schuergers +1
This study engineered Escherichia coli for enhanced extracellular electron transfer (EET) by systematically expressing electron transfer pathways from Shewanella oneidensis MR-1. The complete Mtr pathway (CymA, MtrCAB, and small tetraheme cytochrome STC) achieved the highest electron transfer rates to both soluble acceptors and graphite electrodes. The periplasmic electron shuttle STC proved essential, showing cooperative effects with the Mtr complex. The optimized cymstcmtr strain demonstrated 2–3-fold higher current generation and enhanced methyl orange degradation, positioning engineered E. coli as a versatile platform for microbial electrochemical systems.
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Extracellular electron transfer (EET) engineering in Escherichia coli holds great potential for bioremediation, energy and electrosynthesis applications fueled by readily available organic substrates. Due to its vast metabolic capabilities and availability of synthetic biology tools to adapt strains to specific applications, E. coli is of advantage over native exoelectrogens, but limited in electron transfer rates. We enhanced EET in engineered strains through systematic expression of electron transfer pathways differing in cytochrome composition, localization and origin. While a hybrid pathway harboring components of an E. coli nitrate reductase and the Mtr complex from the exoelectrogen Shewanella oneidensis MR-1 enhanced EET, the highest efficiency was achieved by implementing the complete Mtr pathway from S. oneidensis MR1 in E. coli . We show periplasmic electron shuttling through overexpression of a small tetraheme cytochrome to be central to the electroactivity of this strain, leading to enhanced degradation of the pollutant methyl orange and significantly increased electrical current to graphite electrodes.