Optimizing Escherichia coli's metabolism for fuel cell applications
Date of Completion
Biology, Microbiology|Engineering, Environmental|Energy
In the last few years there have been many publications about applications that center on the generation of electrons from bacterial cells. These applications take advantage of the catabolic diversity of microbes to generate electrical power. The practicality of these applications depends on the microorganism's ability to effectively donate electrons, either directly to the electrode or indirectly through the use of a mediator. After establishing the limitations of electrical output in microbial fuel cells (MFCs) imposed by the bacterial cells, a spectrophotometric assay measuring the indirect reduction of the electronophore neutral red via iron reduction was used to measure electron production from Escherichia coli resting cells. ^ Using this assay I identified NADH dehydrogenase I as a likely site of neutral red reduction. The only previously reported site of interaction between E. coli cells and NR is at the hydrogenases. Although we cannot rule out the possibility that NR is reduced by soluble hydrogenases in the cytoplasm, this previous report indicated that hydrogenase activity does not account for all of the NR reduction activity. Supporting this, data in this thesis suggest that the hydrogenases play a small role in NR reduction. It seems that NR reduction is largely taking place within the cytoplasmic membrane of the bacterial cells, serving as a substrate of enzymes that typically reduce quinones. Furthermore, it seems that under the experimental conditions used here, E. coli's catabolism of glucose is rather inefficient. Instead of using the complete TCA cycle, the bacterial cells are carrying out fermentation, leading to incomplete oxidation of the fuel and low yields of electrons. ^ The results obtained from the TC31 strain suggest that eliminating fermentation pathways to improve NR reduction was the correct approach. Following up on this a new strain was created, KN02, which, in addition to the mutations on strain TC31, lacks acetate kinase activity. ^
Nieves, Ismael U, "Optimizing Escherichia coli's metabolism for fuel cell applications" (2007). Doctoral Dissertations. AAI3276637.