• Login
    View Item 
    •   BEARdocs Home
    • Graduate School
    • Electronic Theses and Dissertations
    • View Item
    •   BEARdocs Home
    • Graduate School
    • Electronic Theses and Dissertations
    • View Item
    JavaScript is disabled for your browser. Some features of this site may not work without it.

    Purification, characterization, and kinetic analysis of various metallo-β-lactamases and discovery of novel inhibitors of these enzymes.

    View/Open
    Dissertation (2.411Mb)
    Permissions Form (307.1Kb)
    Author Permissions (263.1Kb)
    Author Permissions (287.7Kb)
    Author Permissions (290.1Kb)
    Access rights
    Worldwide access.
    Access changed 8/20/19.
    Date
    2014-06-11
    Author
    Schlesinger, Sara Rae.
    Metadata
    Show full item record
    Abstract
    Pathogenic bacteria are rapidly becoming antibiotic resistant, at a rate much faster than the production and FDA approval of new antibiotics that can combat these extreme resistant infections. One way to overcome antibiotic resistance is to design inhibitors that shut down current resistance pathways in order for traditionally useful antibiotics to be reintroduced in combination with the new inhibitor to cure these types of infections. One major contributor to antibiotic resistance are enzymes known as β-lactamases, which render the current most widely prescribed class of antibiotics inactive. This work describes the purification of the enzyme Bla2, a metallo-β-lactamase which confers antibiotic resistance to Bacillus anthracis. Aspects of the enzyme’s activity and stability are investigated to find an optimal setting in which potential inhibitors can be analyzed. Three classes of inhibitors are investigated against this enzyme: compounds containing hydroxamate residues, a compound containing a zinc binding group, and a set of DNA/PNA aptamers previously developed by SELEX technology. The hydroxamate compound N-hydroxy-3-((6-(hydroxyamino)-6-oxohexyl)oxy)benzamide exhibited promising inhibitory activity against Bla2 as well as inhibition of growth of Escherichia coli cells expressing the enzyme. The compound that contains the zinc binding group, known as thiomaltol, inhibits Bla2 more strongly than when employed against other metal containing enzymes and is a reversible, slow-binding inhibitor of Bla2, which may provide a clinical advantage for this compound when co-administered with a β-lactam antibiotic. Also, DNA/PNA aptamers were examined against Bla2 and have some inhibitory activity against the enzyme as well as Bacillus subtilis cells expressing Bla2. In addition to Bla2, a truncated version of the metallo-β-lactamase NDM-1 from Klebsiella pneumoniae was purified and kinetically characterized in order to aid in future inhibitor design of this potent enzyme. In conclusion, various metallo-β-lactamases were successfully purified, characterized, and various classes of inhibitors were investigated against Bla2 which have potential as lead compounds to direct future efforts in novel antibiotic design.
    URI
    http://hdl.handle.net/2104/9103
    Department
    Chemistry and Biochemistry.
    Collections
    • Electronic Theses and Dissertations
    • Theses/Dissertations - Chemistry & Biochemistry

    Copyright © Baylor® University All rights reserved. Legal Disclosures.
    Baylor University Waco, Texas 76798 1-800-BAYLOR-U
    Baylor University Libraries | One Bear Place #97148 | Waco, TX 76798-7148 | 254.710.2112 | Contact: libraryquestions@baylor.edu
    If you find any errors in content, please contact librarywebmaster@baylor.edu
    DSpace software copyright © 2002-2016  DuraSpace
    Contact Us | Send Feedback
    TDL
    Theme by 
    Atmire NV
     

     

    Browse

    All of DSpaceCommunities & CollectionsBy Issue DateAuthorsTitlesSubjectsThis CollectionBy Issue DateAuthorsTitlesSubjects

    My Account

    Login

    Statistics

    View Usage Statistics

    Copyright © Baylor® University All rights reserved. Legal Disclosures.
    Baylor University Waco, Texas 76798 1-800-BAYLOR-U
    Baylor University Libraries | One Bear Place #97148 | Waco, TX 76798-7148 | 254.710.2112 | Contact: libraryquestions@baylor.edu
    If you find any errors in content, please contact librarywebmaster@baylor.edu
    DSpace software copyright © 2002-2016  DuraSpace
    Contact Us | Send Feedback
    TDL
    Theme by 
    Atmire NV