Synthetic <i>Klebsiella pneumoniae</i>‐<i>Shewanella oneidensis</i> Consortium Enables Glycerol‐Fed High‐Performance Microbial Fuel Cells

Abstract

Microbial fuel cell (MFC) is an eco‐friendly bio‐electrochemical sys­tem that uses microorganism as biocatalyst to convert biomass into electricity. Glycerol, as a waste in the biodiesel refinery processes, is an appealing substrate for MFC. Nevertheless, glycerol cannot be utilized as carbon source by well‐known exoelectrogens such as Shewanella oneidensis . Herein, to generate electricity by rapidly harnessing glycerol, the authors rationally constructed a Klebsiella pneumoniae ‐ Shewanella oneidensis microbial consortium to efficiently harvest electricity from glyc­erol, in which K. pneumoniae converted glycerol into lactate, fed to S. oneidensis as carbon source and electron donor. To improve electricity output, the authors systematically engineered the consortium in terms of carbon flux distribution and efficiency of extracellular electron transfer (EET). To direct more carbon flux to lactate biosynthesis in K. pneumoniae , the authors eliminated the ethanol pathway by knocking out the alcohol dehydrogenase gene ( adhE ), and enhanced lactate biosynthesis by heterologously expressing a lactate dehydrogen­ase gene ( ldhD ) from Lactobacillus bulgaricus and a lactate transporter gene ( lldP ) from Escherichia coli . To facilitate EET between S. oneidensis and anode surfaces, a biosynthetic flavins pathway from Bacillus subtilis is introduced into S. oneidensis . The author further optimized the glycerol concentration, thus S. oneidensis could be continuously fed with lactate synthesized from K. pneumoniae at a constant rate. Our glycerol‐fed MFC generated a maximum power density of 19.9 mW/m 2 , significantly higher than that of the wild‐type consor­tium. This work suggested that engineering microbial consortia is an effi­cient strategy to expand the spectrum of usable carbon sources and promote electricity power production in MFCs.