Use this URL to cite or link to this record in EThOS: https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.786539
Title: An investigation of recombinant systems in Escherichia coli engineered for increased electroactivity
Author: Hall, Simon
ISNI:       0000 0004 7971 9869
Awarding Body: University of Sheffield
Current Institution: University of Sheffield
Date of Award: 2018
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Abstract:
Microbial fuel cell technology and other bioelectrochemical systems arose both out of a need to address important global issues, such as the availability of clean water and the need for renewable energy sources, and the opportunity afforded by bacteria capable of utilise extracellular, insoluble metals as electron acceptors as a part of their metabolism. Since then our understanding of extracellular electron transfer has increased dramatically, with electroactivity demonstrably engineered into Escherichia coli, and progress made in a range of synthetic biology applications. This thesis aimed to further explore the limitations and challenges faced when working with these heterologous extracellular electron transfer systems in the interest of progressing the field towards promising applications. In order to overcome the variability between testing apparatus, a single chamber bioelectrochemical cell protocol was developed and presented with comparisons between the strains of various bodies of work. In the process of doing so a previously unobserved electrochemical effect specific to recombinant systems was discovered and investigated, revealing the mediator-like behaviour of the products of the chloramphenicol resistance mechanism. This work went on to present the development of a high throughput dye assay for the screening of electroactivity. This was subsequently used on genomic libraries of Shewanella oneidensis MR-1, a dissimilatory metal reducing bacteria, within E. coli, and a novel heterologous electroactive factor identified. Finally, the sensitivity of recombinant E. coli strains to variations in media within a bioelectrochemical cell was highlighted, prompting further investigation. A global proteomics-based approach was taken to characterising changes within E. coli expressing a heterologous extracellular electron transfer pathway within a bioelectrochemical cell. This recommended avenues for future developments of engineered, electroacive E. coli strains.
Supervisor: Wright, Phillip ; Pandhal, Jagroop Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID: uk.bl.ethos.786539  DOI: Not available
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