Booysen, Brendan Sholto2023-07-192023-07-192023https://hdl.handle.net/10539/35715A dissertation submitted in fulfilment of the requirements for the degree of Master of Science in Molecular and Cell Biology to the Faculty of Science, School of Molecular and Cell Biology, University of the Witwatersrand, Johannesburg, 2022`Worldwide energy shortages are becoming the major factor in curtailing the development of the world economy. Due to this, there has been a sustained increase in research into biofuels such as biodiesel due to the ease of production. However, high amounts of co-produced crude glycerol waste is becoming a critical issue for biodiesel production. A favourable outlet for this crude glycerol waste is its use as a renewable feedstock for microbes such as Escherichia coli. In this scenario, the manufacture of biofuels and other chemicals is done through a bioconversion process. This study aims to use processes such as whole cell biocatalysis and biofilm formation on cheap, gel-like carriers to determine whether Escherichia coli Nissle can use glycerol as a feedstock to produce biodiesel. The study concluded that, although EcN does not produce enough fatty acids to initiate a reaction to produce biodiesel, it can produce enough bioethanol and hydroxide catalyst to convert outside-added oil into biodiesel, at the same rate as the former, with much cleaner glycerol than the conventional process. The introduction of genetic modification to Escherichia coli Nissle to enable increased fatty acid production is an avenue to explore to make this technology more efficient and cost-effective for biodiesel production using Escherichia coli Nissle.enDissertation