LanguageTerm (authority = ISO 639-3:2007); (type = text)
English
Abstract
Glycans or carbohydrates are the most abundant class of biomolecules on the planet, but we are far from elucidating their role in the design and regulation of biological ecosystems spanning from organismal- to protein- level. Nearly every human cell-pathogen interaction and immune system related disorder or disease involves protein-glycan and glycan-glycan interactions at the molecular level. With the increase healthcare costs and greater emphasis by the pharmaceutical industry towards developing more effective biological drugs it becomes imperative to unravel the “glycan code”. Unfortunately, glycan synthesis is not template encoded and is the primary reason for the lack of efficient synthetic tools for widespread application. In nature, glycans are synthesized using membrane associated glycosyl transferases that are associated with several limitations including poor expression, solubility, and the need for expensive donor sugars for large scale synthesis. Alternatively, glycosyl hydrolases (GHs) can be reverse engineered and modulated to synthesize sugar polymers using their inherent transglycosylation property. However, transglycosylation pathway of GHs suffers from low yields because of enzyme’s natural preference towards competing water molecules leading to hydrolysis. Fortunately, protein engineering approaches such as rational engineering and directed evolution can address these challenges to shift the equilibrium towards transglycosylation. In this work, two protein engineering strategies that have been developed that can used engineer glycosyl hydrolysis for efficient glycan synthesis. The first approach is structure guided rational approach where a non-catalytic carbohydrate binding domain is fused to inactive GH family 5 enzymes to rescue the transglycosylation activity. The second approach is high throughput screening development to identify mutant variants that can synthesize glycan using azido-sugars. Here, an azide biosensing toolkit was developed that can be used for directed evolution based glycosynthase engineering. Both these engineering approaches can invariably be applied to many different glycosyl hydrolase families to generate potent transglycosidases.
Subject (authority = local)
Topic
Glycan synthesis
Subject (authority = RUETD)
Topic
Chemical and Biochemical Engineering
RelatedItem (type = host)
TitleInfo
Title
Rutgers University Electronic Theses and Dissertations
Identifier (type = RULIB)
ETD
Identifier
ETD_11479
PhysicalDescription
Form (authority = gmd)
InternetMediaType
application/pdf
InternetMediaType
text/xml
Extent
1 online resource (xx, 116 pages)
Note (type = degree)
Ph.D.
Note (type = bibliography)
Includes bibliographical references
RelatedItem (type = host)
TitleInfo
Title
School of Graduate Studies Electronic Theses and Dissertations
Identifier (type = local)
rucore10001600001
Location
PhysicalLocation (authority = marcorg); (displayLabel = Rutgers, The State University of New Jersey)
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