Towards Electrosynthesis in Shewanella: Energetics of Reversing the Mtr Pathway for Reductive Metabolism

Bioelectrochemical systems rely on microorganisms to link complex oxidation/reduction reactions to electrodes. For example, in Shewanella oneidensis strain MR-1, an electron transfer conduit consisting of cytochromes and structural proteins, known as the Mtr respiratory pathway, catalyzes electron flow from cytoplasmic oxidative reactions to electrodes. Reversing this electron flow to drive microbial reductive metabolism offers a possible route for electrosynthesis of high value fuels and chemicals. We examined electron flow from electrodes into Shewanella to determine the feasibility of this process, the molecular components of reductive electron flow, and what driving forces were required. Addition of fumarate to a film of S. oneidensis adhering to a graphite electrode poised at −0.36 V versus standard hydrogen electrode (SHE) immediately led to electron uptake, while a mutant lacking the periplasmic fumarate reductase FccA was unable to utilize electrodes for fumarate reduction. Deletion of the gene encoding the outer membrane cytochrome-anchoring protein MtrB eliminated 88% of fumarate reduction. A mutant lacking the periplasmic cytochrome MtrA demonstrated more severe defects. Surprisingly, disruption of menC, which prevents menaquinone biosynthesis, eliminated 85% of electron flux. Deletion of the gene encoding the quinone-linked cytochrome CymA had a similar negative effect, which showed that electrons primarily flowed from outer membrane cytochromes into the quinone pool, and back to periplasmic FccA. Soluble redox mediators only partially restored electron transfer in mutants, suggesting that soluble shuttles could not replace periplasmic protein-protein interactions. This work demonstrates that the Mtr pathway can power reductive reactions, shows this conduit is functionally reversible, and provides new evidence for distinct CymA:MtrA and CymA:FccA respiratory units

Catabolic Pathways and Enzymes Involved in Anaerobic Methane Oxidation

Microbes use two distinct catabolic pathways for life with the fuel methane: aerobic methane oxidation carried out by bacteria and anaerobic methane oxidation carried out by archaea. The archaea capable of anaerobic oxidation of methane, anaerobic methanotrophs (ANME), are phylogenetically related to methanogens. While the carbon metabolism in ANME follows the pathway of reverse methanogenesis, the mode of electron transfer from methane oxidation to the terminal oxidant is remarkably versatile. This chapter discusses the catabolic pathways of methane oxidation coupled to the reduction of nitrate, sulfate, and metal oxides. Methane oxidation with sulfate and metal oxides are hypothesized to involve direct interspecies electron transfer and extracellular electron transfer. Cultivation of ANME, their mechanisms of energy conservation, and details about the electron transfer pathways to the ultimate oxidants are rather new and quickly developing research fields, which may reveal novel metabolisms and redox reactions. The second section focuses on the carbon catabolism from methane to CO2 and the biochemistry in ANME with its unique enzymes containing Fe, Ni, Co, Mo, and W that are compared with their homologues found in methanogens

Bacterial Power: An Alternative Energy Source

The demand for energy and the limited supply of fossil fuels and their impact in the environment have required the development of alternative energy sources. Among the next generation of energy sources, microbial fuel cells (MFCs) have emerged as a promising technology due to their ability to recover energy from wastewaters in the form of electricity using electroactive microorganisms as catalysts. Among the various factors that affect power generation performance in MFCs, the efficiency of extracellular electron transfer (EET) is one of the most important. Several enzymes, specifically multiheme cytochromes, have been implicated in this process although the electron transfer chain organization remains to be fully understood. In this chapter, we review in detail the mechanisms that support EET from electroactive microorganisms to the anode in MFCs. We focus on the model organism Shewanella oneidensis MR-1, due to the existence of an extensive molecular characterization of its EET processes. The recent developments in the characterization of the multiheme cytochromes involved in these mechanisms will also be reviewed.authorsversionpublishe