The Genetic Domestication of Clostridium carboxidivorans

The continued use of fossil fuels to produce energy and chemicals is becoming increasingly untenable. They are a finite resource responsible for the release greenhouse gases into the atmosphere, directly contributing to global warming and climate change. If the consumption of fossil fuels remains un...

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Main Author: Morris, William
Format: Thesis (University of Nottingham only)
Language:English
Published: 2025
Subjects:
Online Access:https://eprints.nottingham.ac.uk/81116/
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author Morris, William
author_facet Morris, William
author_sort Morris, William
building Nottingham Research Data Repository
collection Online Access
description The continued use of fossil fuels to produce energy and chemicals is becoming increasingly untenable. They are a finite resource responsible for the release greenhouse gases into the atmosphere, directly contributing to global warming and climate change. If the consumption of fossil fuels remains unabated then drastic environmental consequences are inevitable. A promising alternative is exploiting the potential of microorganisms as biological catalysts to produce renewable energy and chemicals. To this end, autotrophic bacteria that are capable of fixing inorganic carbon in the form of CO2 and CO are of particular interest. Clostridium carboxidivorans is one such bacterium, an anaerobic acetogen capable of producing ethanol, butanol, hexanol, and their conjugate organic acids from CO or CO2 and H2. However, C. carboxidivorans possesses an expansive Restriction Modification System (RMS) rendering DNA transfer impossible. In this study, the RMS of C. carboxidivorans is bypassed to deliver CRISPR-Cas9 vectors targeting RMS-associated nuclease-encoding genes to create a fully genetically domesticated strain, C. carboxidivorans Δ7RM. Then, a metabolic engineering approach is adopted whereby the gene hytA of the domesticated strain is deleted to increase autotrophic growth and ethanol, butanol, and hexanol production. Finally, an alternative Retrotransposition Activated Marker is developed for the ClosTron mutagenesis system. This is characterised in several members of Clostridium, including C. carboxidivorans Δ7RM.
first_indexed 2025-11-14T21:05:09Z
format Thesis (University of Nottingham only)
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institution University of Nottingham Malaysia Campus
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spelling nottingham-811162025-07-24T04:40:17Z https://eprints.nottingham.ac.uk/81116/ The Genetic Domestication of Clostridium carboxidivorans Morris, William The continued use of fossil fuels to produce energy and chemicals is becoming increasingly untenable. They are a finite resource responsible for the release greenhouse gases into the atmosphere, directly contributing to global warming and climate change. If the consumption of fossil fuels remains unabated then drastic environmental consequences are inevitable. A promising alternative is exploiting the potential of microorganisms as biological catalysts to produce renewable energy and chemicals. To this end, autotrophic bacteria that are capable of fixing inorganic carbon in the form of CO2 and CO are of particular interest. Clostridium carboxidivorans is one such bacterium, an anaerobic acetogen capable of producing ethanol, butanol, hexanol, and their conjugate organic acids from CO or CO2 and H2. However, C. carboxidivorans possesses an expansive Restriction Modification System (RMS) rendering DNA transfer impossible. In this study, the RMS of C. carboxidivorans is bypassed to deliver CRISPR-Cas9 vectors targeting RMS-associated nuclease-encoding genes to create a fully genetically domesticated strain, C. carboxidivorans Δ7RM. Then, a metabolic engineering approach is adopted whereby the gene hytA of the domesticated strain is deleted to increase autotrophic growth and ethanol, butanol, and hexanol production. Finally, an alternative Retrotransposition Activated Marker is developed for the ClosTron mutagenesis system. This is characterised in several members of Clostridium, including C. carboxidivorans Δ7RM. 2025-07-24 Thesis (University of Nottingham only) NonPeerReviewed application/pdf en cc_by https://eprints.nottingham.ac.uk/81116/1/Thesis%20-%20William%20Morris%20-%2020213165%20corrections.pdf Morris, William (2025) The Genetic Domestication of Clostridium carboxidivorans. PhD thesis, University of Nottingham. Clostridium carboxidivorans anaerobic acetogen autotrophic bacteria greenhouse gases
spellingShingle Clostridium carboxidivorans
anaerobic acetogen
autotrophic bacteria
greenhouse gases
Morris, William
The Genetic Domestication of Clostridium carboxidivorans
title The Genetic Domestication of Clostridium carboxidivorans
title_full The Genetic Domestication of Clostridium carboxidivorans
title_fullStr The Genetic Domestication of Clostridium carboxidivorans
title_full_unstemmed The Genetic Domestication of Clostridium carboxidivorans
title_short The Genetic Domestication of Clostridium carboxidivorans
title_sort genetic domestication of clostridium carboxidivorans
topic Clostridium carboxidivorans
anaerobic acetogen
autotrophic bacteria
greenhouse gases
url https://eprints.nottingham.ac.uk/81116/