Surface display of redox enzymes in microbial fuel cells

A novel concept for a biofuel cell is presented. Enzyme based fuel cells suffer from enzyme instability when a long time of operation is required. Hence, a system that will continuously produce the biocatalyst needed for the system is necessary. A hybrid of an enzyme-based microbial fuel cell was developed. The redox enzyme glucose oxidase from Aspergillus niger was displayed on the surface of Saccharomyces cerevisiae using the Yeast Surface Display System in a high copy number and as an active enzyme. We have demonstrated its activity both biochemically and electrochemically and observed much higher activity over yeast cells not displaying glucose oxidase as well as over purified glucose oxidase from Aspergillus niger. Further, we were able to construct a biofuel cell, where the anode was comprised of the yeast cells displaying glucose oxidase in the presence of a mediator (methylene blue) and the cathode compartment was comprised of the oxygen reducing enzyme laccase from Trametes versicolor and a redox mediator. Our constructed biofuel cell displayed higher power outputs and current densities than those observed for unmodified yeast and a much longer time of operation in comparison with a similar cell where the anode is comprised of purified glucose oxidase.