INCORPORATION OF FLUORESCENT ANALOGUES IN ISOPRENOID PATHWAYS IN ESCHERICHIA COLI

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George, C. (2018). INCORPORATION OF FLUORESCENT ANALOGUES IN ISOPRENOID PATHWAYS IN ESCHERICHIA COLI. Unc Charlotte Electronic Theses And Dissertations.
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Abstract

Bacterial glycans and glycoconjugates have wide diversity compared to their eukaryotic counterparts. While methods have been developed to monitor and track glycans in eukaryotic systems with some success, there are limited methods with which to do so in prokaryotes. Work conducted by Bertozzi has utilized alkyne-tagged sugars that, once incorporated into enzymatic pathways, can be clicked to a fluorophore for characterization.1 However, due to the increased number of available sugars and potential combinations, this approach this approach becomes challenging in bacteria. Isoprenoids are one of the largest groups of naturally occurring compounds and are essential in all living organisms.2 They are constituted of the building block isopentenyl diphosphate (IPP) and, in prokaryotes, are vital for nearly all polysaccharide biosynthetic pathways via the formation of the anchor bactoprenyl phosphate (BP).3 This anchor is utilized in the cell membrane for a variety of biosynthetic pathways including those related to peptidoglycan (PG), capsular polysaccharides (CPS), exopolysaccharides (EPS), and others.4 This work examines utilizing an exogenously introduced fluorescent isoprenoid to tag and monitor the formation of glycan intermediates in Escherichia coli. This approach has applications in identifying the functions of pertinent enzymes in previously unexamined pathways as well as better understanding associated glycan intermediates. The presented research discusses preliminary methods for incorporation of fluorescent compounds as well as several applications.

Details
Author

George, Chelsea

Title

INCORPORATION OF FLUORESCENT ANALOGUES IN ISOPRENOID PATHWAYS IN ESCHERICHIA COLI

Physical Description

1 online resource (96 pages) : PDF

Date

2018

Degree Granting Institution

University of North Carolina at Charlotte

Abstract

Bacterial glycans and glycoconjugates have wide diversity compared to their eukaryotic counterparts. While methods have been developed to monitor and track glycans in eukaryotic systems with some success, there are limited methods with which to do so in prokaryotes. Work conducted by Bertozzi has utilized alkyne-tagged sugars that, once incorporated into enzymatic pathways, can be clicked to a fluorophore for characterization.1 However, due to the increased number of available sugars and potential combinations, this approach this approach becomes challenging in bacteria. Isoprenoids are one of the largest groups of naturally occurring compounds and are essential in all living organisms.2 They are constituted of the building block isopentenyl diphosphate (IPP) and, in prokaryotes, are vital for nearly all polysaccharide biosynthetic pathways via the formation of the anchor bactoprenyl phosphate (BP).3 This anchor is utilized in the cell membrane for a variety of biosynthetic pathways including those related to peptidoglycan (PG), capsular polysaccharides (CPS), exopolysaccharides (EPS), and others.4 This work examines utilizing an exogenously introduced fluorescent isoprenoid to tag and monitor the formation of glycan intermediates in Escherichia coli. This approach has applications in identifying the functions of pertinent enzymes in previously unexamined pathways as well as better understanding associated glycan intermediates. The presented research discusses preliminary methods for incorporation of fluorescent compounds as well as several applications.

Genre

masters theses

Subjects--Topics

Biochemistry

Degree

M.S.

Subject Area

Chemistry

Advisor(s)

Troutman, Jerry

Committee Members

Afonin, Kirill
Vivero-Escoto, Juan
Steck, Todd

Degree Note

Thesis (M.S.)--University of North Carolina at Charlotte, 2018.

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Rights Holder Information

Copyright is held by the author unless otherwise indicated.

Identifier

George_uncc_0694N_11827

Permalink

http://hdl.handle.net/20.500.13093/etd:30