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Tools to explore carbohydrate metabolism pathways in Mycobacteria
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Parker, Hadyn Luke (2021) Tools to explore carbohydrate metabolism pathways in Mycobacteria. PhD thesis, University of Warwick.
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Official URL: http://webcat.warwick.ac.uk/record=b3764393
Abstract
Mycobacterium tuberculosis (Mtb) survives intracellularly in a nutrient deprived environment, within the human host, for decades. The nutrients that Mtb uses to survive are not well understood and, in particular, the pathways that are involved in carbohydrate uptake and metabolism are not well defined.
Mtb has a highly unique cell-wall comprising of complex carbohydrates and lipids. Trehalose, a disaccharide, is an essential component of trehalose-lipids within the Mtb cell-wall, which play a key role in virulence. Recent studies have shown that trehalose provides an energy source to Mtb through its uptake and recycling by the LpqY-SugABC transporter before being metabolised through various pathways.
To further understand how Mtb utilises trehalose, novel ‘unnatural’ trehalose analogues were synthesised to evaluate how modifications might alter uptake, toxicity and stability. Four trehalose analogues; mannotrehalose (ManGlc), galactotrehalose (GalGlc) and 6-azido derivatives were chemoenzymatically synthesised by trehalose synthase (TreT). The trehalose analogues were found to be imported into the cell and potentially incorporated into the cell envelope in M. smegmatis and M. bovis BCG. The analogues are well tolerated by mycobacteria as they did not inhibit mycobacterial growth or influence trehalose processing proteins within the proteome. However, the trehalose analogues were resistant to utilisation by trehalase, TreS and OtsA, the main trehalose processing pathways in mycobacteria, and therefore could not be used to support growth in minimal media. Combined these results present novel insights into how trehalose is processed by mycobacteria and provide the tools for enzymatically stable, trehalose-based probes with potential applications in the diagnostic or treatment of Mtb.
In addition, the differential radial capillary action of ligand assay was optimised for the high-throughput detection of carbohydrate binding proteins. The development of an Mtb overexpression ORFeome is ongoing and could be used to identify novel trehalose binding proteins or pathways in mycobacteria.
| Item Type: | Thesis (PhD) | ||||
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| Subjects: | Q Science > QH Natural history Q Science > QP Physiology Q Science > QR Microbiology |
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| Library of Congress Subject Headings (LCSH): | Mycobacterium tuberculosis, Mycobacteria -- Metabolism, Carbohydrates -- Metabolism, Trehalose | ||||
| Official Date: | March 2021 | ||||
| Dates: |
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| Institution: | University of Warwick | ||||
| Theses Department: | School of Life Sciences | ||||
| Thesis Type: | PhD | ||||
| Publication Status: | Unpublished | ||||
| Supervisor(s)/Advisor: | Fullam, Elizabeth ; Gibson, Matthew I. | ||||
| Sponsors: | Midlands Integrative Biosciences Training Partnership ; Biotechnology and Biological Sciences Research Council (Great Britain) | ||||
| Format of File: | |||||
| Extent: | xxiv, 319 leaves : illustrations | ||||
| Language: | eng |
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