Chemoenzymatic Preparation of Probes for in vitro and Cellular Elucidation of Complex Bacteria Polysaccharides

Search for this publication on Google Scholar
Reid, A. (2021). Chemoenzymatic Preparation of Probes for in vitro and Cellular Elucidation of Complex Bacteria Polysaccharides. Unc Charlotte Electronic Theses And Dissertations.
Download PDF

Analytics

35 views ◎
13 downloads ⇓


Abstract

Bacteria play a major role in our health and wellbeing. The microbe-host interaction is often mediated by sugar polymers at the cell surface. An incredibly diverse amount of glycan variation exists throughout these structures, which makes identification of surface components difficult. The composition of the surface is unique to bacteria and acts as molecular fingerprint which can distinguish even subspecies apart. Better methods to decipher what those glycan identities are or how to reproduce them may help develop future advancements towards exploiting them as therapeutic targets. The major challenge addressed herein is to simplify the tools used to track the formation of these natural materials. To do this, we expand on the chemoenzymatic preparation of a tagged lipid substrate central to early stages of biosynthesis for many surface polysaccharides. Further, we identify conditions in which these unnatural substrates can be used in vitro. Lastly, we develop methodologies to detect BP and polysaccharide intermediates in live cells. These tools have facilitated robust detection and reconstruction of glycan assembly building blocks. This research may lay the ground work for future applications technologies towards novel therapeutics, such as glycoconjugates vaccines.

Details
Author

Reid, Amanda

Title

Chemoenzymatic Preparation of Probes for in vitro and Cellular Elucidation of Complex Bacteria Polysaccharides

Physical Description

1 online resource (151 pages) : PDF

Date

2021

Degree Granting Institution

University of North Carolina at Charlotte

Abstract

Bacteria play a major role in our health and wellbeing. The microbe-host interaction is often mediated by sugar polymers at the cell surface. An incredibly diverse amount of glycan variation exists throughout these structures, which makes identification of surface components difficult. The composition of the surface is unique to bacteria and acts as molecular fingerprint which can distinguish even subspecies apart. Better methods to decipher what those glycan identities are or how to reproduce them may help develop future advancements towards exploiting them as therapeutic targets. The major challenge addressed herein is to simplify the tools used to track the formation of these natural materials. To do this, we expand on the chemoenzymatic preparation of a tagged lipid substrate central to early stages of biosynthesis for many surface polysaccharides. Further, we identify conditions in which these unnatural substrates can be used in vitro. Lastly, we develop methodologies to detect BP and polysaccharide intermediates in live cells. These tools have facilitated robust detection and reconstruction of glycan assembly building blocks. This research may lay the ground work for future applications technologies towards novel therapeutics, such as glycoconjugates vaccines.

Genre

doctoral dissertations

Subjects--Topics

Microbiology
Biochemistry
Analytical chemistry

Degree

Ph.D.

Keywords

Bactoprenyl Diphosphate
Glycan Immobilization
Initiating Glycosyltransferases
Lipid Linked Oligosaccharides
Polyisoprenoids
Undecaprenyl Diphosphate Synthase

Subject Area

Nanoscale Science

Advisor(s)

Troutman, Jerry

Committee Members

Ogle, Craig
Nesmelov, Yuri
De Silva Indrasakara, Swarnapali
Clemens, Mark

Degree Note

Thesis (Ph.D.)--University of North Carolina at Charlotte, 2021.

Rights Statement

This Item is protected by copyright and/or related rights. You are free to use this Item in any way that is permitted by the copyright and related rights legislation that applies to your use. For other uses you need to obtain permission from the rights-holder(s). For additional information, see http://rightsstatements.org/page/InC/1.0/.

Rights Holder Information

Copyright is held by the author unless otherwise indicated.

Identifier

Reid_uncc_0694D_12676

Permalink

http://hdl.handle.net/20.500.13093/etd:2052