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dc.contributor.authorSimon, David
dc.contributor.otherQueen's University (Kingston, Ont.). Theses (Queen's University (Kingston, Ont.))en
dc.date.accessioned2019-09-04T21:30:21Z
dc.date.available2019-09-04T21:30:21Z
dc.identifier.urihttp://hdl.handle.net/1974/26512
dc.description.abstractThe Streptomyces are a genus of soil-dwelling Gram-positive bacteria that produce a variety of secondary metabolites; they have even been dubbed as ‘antibiotic factories’ as a typical genome can contain as many as 30 clusters of genes responsible for biosynthesis of metabolites. Yet many of these compounds are not observed in laboratory cultures suggesting the products are locked by regulatory signals. One unique regulatory mechanism present in these organisms, the bldA gene, depends on the translation TTA codons within the genome. These codons are quite rare but are frequently found in other regulatory regions and in regions responsible for the biosynthesis of natural products. This thesis applies mass spectrometry to explore changes in the whole proteome of Streptomyces calvus in response to the bldA gene. S. calvus pTESa-bldA can undergo morphological differentiation and produces different secondary metabolites than the native strain. Hypothesized to be due to the action of a TTA codon containing pleotropic regulator AdpA, we observed that AdpA is expressed in the wild type at the protein level. Additionally, there was an increase in expression of other regulatory systems in reaction to bldA including the two-component system regulators controlling the phosphate management system and MtrAB system known to control cell division. Both systems have been previously demonstrated to have an impact on secondary metabolism and morphological differentiation. Furthermore, a combination of bioinformatics and high-resolution mass spectrometry were used to identify and guide purification of two natural products from Streptomyces curacoi in response to bldA. A non-ribosomal peptide, curacomycin, and a ribosomally synthesized and post-translationally modified peptide, curacozole, both were isolated and characterized from S. curacoi pTESa-bldA. Curacozole was produced with a high titre, 42 mg/L, that allowed for extensive antibacterial testing where it was found to be highly active against antibiotic resistant Gram-positive bacteria. Lastly, custom electrospray emitters were fabricated with significant signal enhancement to aid mass spectrometric investigations of these biological systems.en_US
dc.language.isoenen_US
dc.relation.ispartofseriesCanadian thesesen
dc.rightsCC0 1.0 Universal*
dc.rightsQueen's University's Thesis/Dissertation Non-Exclusive License for Deposit to QSpace and Library and Archives Canadaen
dc.rightsProQuest PhD and Master's Theses International Dissemination Agreementen
dc.rightsIntellectual Property Guidelines at Queen's Universityen
dc.rightsCopying and Preserving Your Thesisen
dc.rightsThis publication is made available by the authority of the copyright owner solely for the purpose of private study and research and may not be copied or reproduced except as permitted by the copyright laws without written authority from the copyright owner.en
dc.rights.urihttp://creativecommons.org/publicdomain/zero/1.0/*
dc.subjectNatural Productsen_US
dc.subjectMass Spectrometryen_US
dc.subjectBiosynthesisen_US
dc.subjectProteomicsen_US
dc.subjectGenomicsen_US
dc.titleApplications of Mass Spectrometry in the Studies Concerning bldA Regulation of Natural Product Biosynthesis in Streptomyces sp.en_US
dc.typethesisen
dc.description.degreeDoctor of Philosophyen_US
dc.contributor.supervisorZechel, David
dc.contributor.supervisorOleschuk, Richard
dc.contributor.departmentChemistryen_US
dc.embargo.termsThis thesis contains unpublished findings in a competitive field. We are in the process of completing the manuscript for future publication.en_US
dc.embargo.liftdate2024-09-03T18:59:44Z


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