Metabolite-based mutualism between Pseudomonas aeruginosa PA14 and Enterobacter aerogenes enhances current generation in bioelectrochemical systems

Random transposon insertion libraries have proven invaluable in studying bacterial genomes. Libraries that approach saturation must be large, with multiple insertions per gene, making comprehensive genome-wide scanning difficult. To facilitate genome-scale study of the opportunistic human pathogen Pseudomonas aeruginosa strain PA14, we constructed a nonredundant library of PA14 transposon mutants (the PA14NR Set) in which nonessential PA14 genes are represented by a single transposon insertion chosen from a comprehensive library of insertion mutants. The parental library of PA14 transposon insertion mutants was generated by using MAR2xT7, a transposon compatible with transposon-site hybridization and based on mariner. The transposon-site hybridization genetic footprinting feature broadens the utility of the library by allowing pooled MAR2xT7 mutants to be individually tracked under different experimental conditions. A public, internet-accessible database (the PA14 Transposon Insertion Mutant Database, http://ausubellab.mgh.harvard.edu/cgi-bin/pa14/home.cgi) was developed to facilitate construction, distribution, and use of the PA14NR Set. The usefulness of the PA14NR Set in genome-wide scanning for phenotypic mutants was validated in a screen for attachment to abiotic surfaces. Comparison of the genes disrupted in the PA14 transposon insertion library with an independently constructed insertion library in P. aeruginosa strain PAO1 provides an estimate of the number of P. aeruginosa essential genes.

New sustainable methods are needed to produce renewable energy carriers that can be stored and used for transportation, heating, or chemical production. Here we demonstrate that methane can directly be produced using a biocathode containing methanogens in electrochemical systems (abiotic anode) or microbial electrolysis cells (MECs; biotic anode) by a process called electromethanogenesis. At a set potential of less than -0.7 V (vs Ag/AgCl), carbon dioxide was reduced to methane using a two-chamber electrochemical reactor containing an abiotic anode, a biocathode, and no precious metal catalysts. At -1.0 V, the current capture efficiency was 96%. Electrochemical measurements made using linear sweep voltammetry showed that the biocathode substantially increased current densities compared to a plain carbon cathode where only small amounts of hydrogen gas could be produced. Both increased current densities and very small hydrogen production rates by a plain cathode therefore support a mechanism of methane production directly from current and not from hydrogen gas. The biocathode was dominated by a single Archaeon, Methanobacterium palustre. When a current was generated by an exoelectrogenic biofilm on the anode growing on acetate in a single-chamber MEC, methane was produced at an overall energy efficiency of 80% (electrical energy and substrate heat of combustion). These results show that electromethanogenesis can be used to convert electrical current produced from renewable energy sources (such as wind, solar, or biomass) into a biofuel (methane) as well as serving as a method for the capture of carbon dioxide.