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Development of a method to generate a soluble substrate for lytic transglycosylases

Peptidoglycan, the major component of the bacterial cell wall, is essential for cell viability. Several important antibiotics disrupt peptidoglycan metabolism, including the β-lactams and vancomycin. There are several bacterial enzymes involved in peptidoglycan metabolism that are not yet the target of antibiotics, such as the lytic transglycosylases (LTs). Relatively little experimental characterization has been done on LTs, due largely to the difficulties of working with insoluble, heterogeneous, and highly variable peptidoglycan. This research develops a method for the generation of a soluble, homogeneous oligosaccharide substrate that can be used to study LTs. The approach taken was based on the enzymatic degradation of peptidoglycan into fragments of a specific nature, and their separation by HPLC. This work identifies the challenges associated with this approach, and discusses the potential flaws in the 'top-down' generation of a soluble substrate. / This thesis was typeset with LaTeX using Minion Pro and Myriad Pro typefaces.

Identiferoai:union.ndltd.org:LACETR/oai:collectionscanada.gc.ca:OGU.10214/2644
Date18 April 2011
CreatorsMark, Adam L.
ContributorsClarke, Anthony J.
Source SetsLibrary and Archives Canada ETDs Repository / Centre d'archives des thèses électroniques de Bibliothèque et Archives Canada
LanguageEnglish
Detected LanguageEnglish
TypeThesis
Rightshttp://creativecommons.org/licenses/by/3.0/

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