Tuning the Substrate Specificity of the Glutathione Transferase GstB from <i>Escherichia coli</i> via Site-directed Mutagenesis

Tuning the Substrate Specificity of the Glutathione Transferase GstB from <i>Escherichia coli</i> via Site-directed Mutagenesis

Bibliographic Details
Main Author: Moore, Jennifer Marie
Language:English
Published: Youngstown State University / OhioLINK 2017
Subjects:
Biochemistry
Chemistry
Microbiology
Glutathione
glutathione transferases
site-directed mutagenesis
bioremediation
bromoacetate
Online Access:http://rave.ohiolink.edu/etdc/view?acc_num=ysu1502910733353098
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spelling ndltd-OhioLink-oai-etd.ohiolink.edu-ysu15029107333530982021-08-03T07:03:57Z Tuning the Substrate Specificity of the Glutathione Transferase GstB from <i>Escherichia coli</i> via Site-directed Mutagenesis Moore, Jennifer Marie Biochemistry Chemistry Microbiology Glutathione glutathione transferases site-directed mutagenesis bioremediation bromoacetate Glutathione transferases (GSTs) are detoxication enzymes that are widely distributed in nature. They fulfill their protective roles by catalyzing the conjugation of the tripeptide glutathione to both endogenous and xenobiotic electrophiles. Eukaryotic GSTs are the subject of thorough investigation, while the prokaryotic enzymes remain relatively unexplored. GstB is a glutathione transferase from <i>Escherichia coli</i> that is known to detoxify bromoacetate, a water disinfection by-product. This work has served to expand the substrate scope for GstB. Site-directed mutagenesis of the electrophile binding site residue arginine 119 was performed, generating the alanine, glutamine, histidine, and serine enzyme variants. The activities of the mutants toward a range of electrophiles were evaluated to investigate the impact of the amino acid substitutions on substrate specificity. Initial activity screening results indicate that some mutants display rate enhancement for acrylate and iodoacetamide conjugation. Kinetic parameters with iodoacetamide suggest that certain mutants are more catalytically efficient and resistant to inhibition compared to the wild type enzyme. The finding that amino acid substitution at position 119 can modify GstB substrate specificity provides support for pollutant-targeted bioremediation strategies. 2017-08-17 English text Youngstown State University / OhioLINK http://rave.ohiolink.edu/etdc/view?acc_num=ysu1502910733353098 http://rave.ohiolink.edu/etdc/view?acc_num=ysu1502910733353098 unrestricted This thesis or dissertation is protected by copyright: all rights reserved. It may not be copied or redistributed beyond the terms of applicable copyright laws.
collection NDLTD
language English
sources NDLTD
topic Biochemistry
Chemistry
Microbiology
Glutathione
glutathione transferases
site-directed mutagenesis
bioremediation
bromoacetate
spellingShingle Biochemistry
Chemistry
Microbiology
Glutathione
glutathione transferases
site-directed mutagenesis
bioremediation
bromoacetate
Moore, Jennifer Marie
Tuning the Substrate Specificity of the Glutathione Transferase GstB from <i>Escherichia coli</i> via Site-directed Mutagenesis
author Moore, Jennifer Marie
author_facet Moore, Jennifer Marie
author_sort Moore, Jennifer Marie
title Tuning the Substrate Specificity of the Glutathione Transferase GstB from <i>Escherichia coli</i> via Site-directed Mutagenesis
title_short Tuning the Substrate Specificity of the Glutathione Transferase GstB from <i>Escherichia coli</i> via Site-directed Mutagenesis
title_full Tuning the Substrate Specificity of the Glutathione Transferase GstB from <i>Escherichia coli</i> via Site-directed Mutagenesis
title_fullStr Tuning the Substrate Specificity of the Glutathione Transferase GstB from <i>Escherichia coli</i> via Site-directed Mutagenesis
title_full_unstemmed Tuning the Substrate Specificity of the Glutathione Transferase GstB from <i>Escherichia coli</i> via Site-directed Mutagenesis
title_sort tuning the substrate specificity of the glutathione transferase gstb from <i>escherichia coli</i> via site-directed mutagenesis
publisher Youngstown State University / OhioLINK
publishDate 2017
url http://rave.ohiolink.edu/etdc/view?acc_num=ysu1502910733353098
work_keys_str_mv AT moorejennifermarie tuningthesubstratespecificityoftheglutathionetransferasegstbfromiescherichiacoliiviasitedirectedmutagenesis
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