01whole.pdf (13.67 MB)
An enzyme for nucleotide sugar modification in A. Baumannii
thesisposted on 2022-03-28, 11:08 authored by Heather Clift
The gram-negative bacillus Acinetobacter baumannii is an emerging multidrug-resistant, opportunistic pathogen responsible for severe nosocomial and community pneumonia outbreaks worldwide. Increasing resistance to antibiotics is thought to be due in part to highly variable genomic islands and lateral gene transfer. To date there have been studies into multiple genes in A. baumannii that are thought to play major roles in its resistance capabilities. One such gene found in a genomic island of the D1279779 strain of A. baumannii, fnIA, encodes for the protein Ab-WbjB, an extended short-chain dehydrogenase/reductase (SDR). Although it is similar in structure to other members of the SDR family, little is known about its function in A. baumannii. I have carried out analysis on the active site chemistry of Ab-WbjB, discovering the necessity for the presence of its cofactor NADP for both overall protein stability and substrate binding. I have also found that Ab-WbjB binds to the substrate analogue UDP-GlcNAc. In addition to functional assays of the protein, an attempt at crystal growth was made under conditions containing both NADP and UDP-GlcNAc. A knockout of the fnlA gene in A. baumannii was also planned, but has yet to be completed.
Table of ContentsChapter 1. Introduction -- Chapter 2. Materials and methods -- Chapter 3. Production and buffer characterisation of recombinant forms of Ab-WbjB -- Chapter 4. Biochemical annotation of Ab-WbjB -- Chapter 5. Acinetobacter baumannii gene knockout -- Chapter 6. Conclusions and future directions.
NotesBibliography: pages a-f Empirical thesis.
Awarding InstitutionMacquarie University
Degree TypeThesis MRes
DegreeMRes, Macquarie University, Faculty of Science and Engineering, Department of Chemistry and Biomolecular Sciences
Department, Centre or SchoolDepartment of Chemistry and Biomolecular Sciences
Year of Award2014
Principal SupervisorBridget Mabbutt
RightsCopyright Heather Clift 2014. Copyright disclaimer: http://mq.edu.au/library/copyright
Extent1 online resource (iv, 54, a-f pages) illustrations (some colour)
Former Identifiersmq:53727 http://hdl.handle.net/1959.14/1137098