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In search of an effective inhibitor of glutamate racemase Yu, Warrick Sze Chung
Abstract
Glutamate racemase is an enzyme involved in the biosynthesis of peptidoglycan in bacteria. Designing an effective inhibitor of this enzyme can lead to a better understanding of the mechanism employed by this enzyme as well as the development of new antibiotics. A potential inhibitor, 3-(l-hydroxy-2-imidazolyl)propanoic acid, 1, was designed. This compound was synthesized via the construction of an Nhydroxyimidazole ring and the attachment of the glutamate side chain to Position 2 of the ring. A well-established procedure was used in the construction of the imidazole ring involving condensation of hydroxylamine, glyoxal and formaldehyde. However, the attachment of the glutamate side chain involved the introduction of an aldehyde group at Position 2 of the ring and then the attachment of the rest of the side chain via Wittig reaction. Finally, hydrogenation removed the benzyl protecting groups and yielded the desired compound. The potential inhibitor, 1, was then tested with glutamate racemase in a coupled spectrophotometric assay, which involved the use of L-glutamate dehydrogenase and diaphorase as the coupling enzymes. It was found that Compound 1 was a good competitive inhibitor of the coupling enzyme, L-glutamate dehydrogenase, but not of glutamate racemase. The K₁ value of 1 towards L-glutamate dehydrogenase was found to be 1.66 ± 0.52 mM. However, the precise K₁ value towards glutamate racemase was not pursued due to ineffective inhibition. It was suggested that the poor inhibition of 1 towards glutamate racemase was caused by two possible factors. First, the absence of essential charged groups required for substrate binding to the glutamate racemase active site might lead to poor binding of Compound 1. Second, the bulkiness of the imidazole ring might cause steric problems such that the inhibitor could not fit into the glutamate racemase active site.
Item Metadata
Title |
In search of an effective inhibitor of glutamate racemase
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Creator | |
Publisher |
University of British Columbia
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Date Issued |
2000
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Description |
Glutamate racemase is an enzyme involved in the biosynthesis of peptidoglycan
in bacteria. Designing an effective inhibitor of this enzyme can lead to a better
understanding of the mechanism employed by this enzyme as well as the development of
new antibiotics. A potential inhibitor, 3-(l-hydroxy-2-imidazolyl)propanoic acid, 1, was
designed. This compound was synthesized via the construction of an Nhydroxyimidazole
ring and the attachment of the glutamate side chain to Position 2 of the
ring. A well-established procedure was used in the construction of the imidazole ring
involving condensation of hydroxylamine, glyoxal and formaldehyde. However, the
attachment of the glutamate side chain involved the introduction of an aldehyde group at
Position 2 of the ring and then the attachment of the rest of the side chain via Wittig
reaction. Finally, hydrogenation removed the benzyl protecting groups and yielded the
desired compound.
The potential inhibitor, 1, was then tested with glutamate racemase in a coupled
spectrophotometric assay, which involved the use of L-glutamate dehydrogenase and
diaphorase as the coupling enzymes. It was found that Compound 1 was a good
competitive inhibitor of the coupling enzyme, L-glutamate dehydrogenase, but not of
glutamate racemase. The K₁ value of 1 towards L-glutamate dehydrogenase was found to
be 1.66 ± 0.52 mM. However, the precise K₁ value towards glutamate racemase was not
pursued due to ineffective inhibition. It was suggested that the poor inhibition of 1
towards glutamate racemase was caused by two possible factors. First, the absence of
essential charged groups required for substrate binding to the glutamate racemase active
site might lead to poor binding of Compound 1. Second, the bulkiness of the imidazole
ring might cause steric problems such that the inhibitor could not fit into the glutamate
racemase active site.
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Extent |
12565671 bytes
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Genre | |
Type | |
File Format |
application/pdf
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Language |
eng
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Date Available |
2009-07-20
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Provider |
Vancouver : University of British Columbia Library
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Rights |
For non-commercial purposes only, such as research, private study and education. Additional conditions apply, see Terms of Use https://open.library.ubc.ca/terms_of_use.
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DOI |
10.14288/1.0061496
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URI | |
Degree | |
Program | |
Affiliation | |
Degree Grantor |
University of British Columbia
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Graduation Date |
2000-11
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Campus | |
Scholarly Level |
Graduate
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Aggregated Source Repository |
DSpace
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Item Media
Item Citations and Data
Rights
For non-commercial purposes only, such as research, private study and education. Additional conditions apply, see Terms of Use https://open.library.ubc.ca/terms_of_use.