Discovery of Staphylococcus aureus bacteria cell division FtsZ protein inhibitors via STD NMR

Badal, T

Discovery of Staphylococcus aureus bacteria cell division FtsZ protein inhibitors via STD NMR

Badal, T

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Publication Type:: Thesis
Issue Date:: 2014

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Field	Value	Language
dc.contributor.author	Badal, T
dc.date.accessioned	2015-03-27T04:50:18Z
dc.date.available	2015-03-27T04:50:18Z
dc.date.issued	2014
dc.identifier.uri	http://hdl.handle.net/10453/34481
dc.description	University of Technology Sydney. Faculty of Science.	en_US
dc.description	NO FULL TEXT AVAILABLE. This thesis contains 3rd party copyright material. The hardcopy may be available for consultation at the UTS Library.
dc.description.abstract	NO FULL TEXT AVAILABLE. This thesis contains 3rd party copyright material. ----- The increase of antibiotic resistance among an array of bacteria has engendered a search for antibiotics with novel modes of action. Pathogens such as Methicillin-resistant Staphylococcus aureus (MRSA) have shown resistance towards every class of antibiotics available and are of growing concern in both hospital and community settings. Hence, targeting bacterial cell division proteins is a new approach in developing new antibiotics. One such emerging target is FtsZ, a cell division protein that is highly conserved among bacteria and found to be an essential protein in 1990. Although berberine has been shown to inhibit FtsZ in Escherichia coli, optimization of this drug was necessary. Thus, this thesis further investigates the potential of using natural products to inhibit bacterial cell division such as berberine, eugenol, cinnamaldehyde, and synthesised berberine-like molecules by targeting S. aureus FtsZ (SaFtsZ). Berberine-like molecules, 4 and 4a were successfully synthesised and their inhibitory activity analysed via Saturation Transfer Difference Nuclear Magnetic Resonance (STD NMR) and minimum inhibitory concentrations (MIC). STD NMR standard operation procedures were first developed and optimised for implementation on an Agilent 500 MHz NMR by way of a model study using human serum albumin and tryptophan as the protein and ligand, respectively. SaFtsZ was overexpressed and purified for STD NMR studies and the procedures were successfully applied in the investigation of SaFtsZ-ligand interactions. Following comparative analysis with molecular modelling, compounds with benzene conjugated systems and nitro substituted benzene rings were found to improve SaFtsZ binding affinity. Biological assays of the five compounds were utilised to assess the potency of each compound. This was accomplished via MIC assays to determine the lowest concentration required to inhibit the growth of S. aureus ATCC25923, and berberine was found to have MIC value of 128 μg/mL, two serial dilutions less than MIC values of eugenol, cinnamaldehyde, 4 and 4a.	en_US
dc.format	Thesis (MSc)	en_US
dc.language.iso	en	en_US
dc.rights	The author owns the copyright in this thesis including all reproduction and reuse rights for the work. The work may not be altered without the permission of the copyright owner. Attribution is essential when quoting or paraphrasing from this thesis.
dc.rights	info:eu-repo/semantics/closedAccess
dc.title	Discovery of Staphylococcus aureus bacteria cell division FtsZ protein inhibitors via STD NMR	en_US
dc.type	Thesis
utslib.copyright.status	closed_access

Abstract:

NO FULL TEXT AVAILABLE. This thesis contains 3rd party copyright material. ----- The increase of antibiotic resistance among an array of bacteria has engendered a search for antibiotics with novel modes of action. Pathogens such as Methicillin-resistant Staphylococcus aureus (MRSA) have shown resistance towards every class of antibiotics available and are of growing concern in both hospital and community settings. Hence, targeting bacterial cell division proteins is a new approach in developing new antibiotics. One such emerging target is FtsZ, a cell division protein that is highly conserved among bacteria and found to be an essential protein in 1990. Although berberine has been shown to inhibit FtsZ in Escherichia coli, optimization of this drug was necessary. Thus, this thesis further investigates the potential of using natural products to inhibit bacterial cell division such as berberine, eugenol, cinnamaldehyde, and synthesised berberine-like molecules by targeting S. aureus FtsZ (SaFtsZ). Berberine-like molecules, 4 and 4a were successfully synthesised and their inhibitory activity analysed via Saturation Transfer Difference Nuclear Magnetic Resonance (STD NMR) and minimum inhibitory concentrations (MIC). STD NMR standard operation procedures were first developed and optimised for implementation on an Agilent 500 MHz NMR by way of a model study using human serum albumin and tryptophan as the protein and ligand, respectively. SaFtsZ was overexpressed and purified for STD NMR studies and the procedures were successfully applied in the investigation of SaFtsZ-ligand interactions. Following comparative analysis with molecular modelling, compounds with benzene conjugated systems and nitro substituted benzene rings were found to improve SaFtsZ binding affinity. Biological assays of the five compounds were utilised to assess the potency of each compound. This was accomplished via MIC assays to determine the lowest concentration required to inhibit the growth of S. aureus ATCC25923, and berberine was found to have MIC value of 128 μg/mL, two serial dilutions less than MIC values of eugenol, cinnamaldehyde, 4 and 4a.

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http://hdl.handle.net/10453/34481