Investigating methylcitric acid cycle enzymes in Bacillus subtilis and polyketide synthase enzymes in Bacillus amyloliquefaciens
- UNCG Author/Contributor (non-UNCG co-authors, if there are any, appear on document)
- Victoria Gail Meadows (Creator)
- Institution
- The University of North Carolina at Greensboro (UNCG )
- Web Site: http://library.uncg.edu/
- Advisor
- Jason Reddick
Abstract: Bacterial secondary metabolites from Bacillus species have an important role in drug discovery and development. This dissertation focuses on research to characterize two enzymes from the Bacillus species: Bacillus subtilis and Bacillus amyloliquefaciens and their biosynthetic pathways. The first chapter of this dissertation will focus on broad topics of understanding secondary metabolites and enzymology of Bacillus species. The second chapter will focus on the enzyme 2-methylcitrate dehydratase, MmgE, from the methylcitric acid cycle of B. subtilis. This enzyme is linked to energy production when the bacteria go through the process of sporulation due to stress. The enzyme was tested for substrate tolerance with the prediction that it could be used in bioorganic chemistry to add chiral alcohol groups to enoyl acids, and the dehydration of the beta-hydroxyacids to form olefin products with specific double bond geometries. The enzyme, MmgE, was shown to have limited substrate tolerance outside of the natural substrates. The maximum velocity rate for MmgE with malic acid is ????????=0.00149±2.009 ??????????. The ????=2.78±0.58 ????. Comparability, MmgE with citric acid is ????????=0.051±0.007 ??????????. The ????=0.186±0.075 ????. The third and last chapter will focus on characterization of the cytochrome P450 enzyme, PksS, from the biosynthetic pathway of the polyketide bacillaene. The trans-acyltransferase polyketide synthase product, bacillaene, is used as an antibiotic and antifungal in the biofilm of the bacteria against predators in the surrounding area. We made attempts to knockout baeS/pksS in their respective Bacillus species to determine the product of the reaction between the knockout mutations and the cytochrome P450 enzyme, PksS. [This abstract may have been edited to remove characters that will not display in this system. Please see the PDF for the full abstract.]
Investigating methylcitric acid cycle enzymes in Bacillus subtilis and polyketide synthase enzymes in Bacillus amyloliquefaciens
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Created on 12/1/2022
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Additional Information
- Publication
- Dissertation
- Language: English
- Date: 2022
- Keywords
- Bacillus amyloliquefaciens, Bacillus subtilis, Cytochrome P450, Methylcitric acid cycle, Polyketide Synthase
- Subjects
- Microbial metabolites
- Enzymology
- Bacillus (Bacteria)