{"id":{"repo_id":"uiuc","oai_identifier":"oai:www.ideals.illinois.edu:2142/89125"},"canonical_url":"https://search.dev.ndltd.org/etd/uiuc/oai:www.ideals.illinois.edu:2142/89125","repository":{"repo_id":"uiuc","name":"University of Illinois - Urbana-Champaign","base_url":"https://www.ideals.illinois.edu/oai-pmh"},"display":{"title":"Structure and function analysis of the natural product plantazolicin, a bacillus anthracis-specific antibiotic","abstract":"Bacteria are a fruitful source of metabolites, many of which have been the scaffolds for the majority of approved antibiotic compounds. In this dissertation, I present the discovery and characterization of a bacterial natural product from Bacillus methylotrophicus FZB42, a Gram-positive, rod-shaped bacterium that stimulates plant growth. A prolific producer of secondary metabolites, FZB42 excretes a compound bearing the molecular mass of 1335 Daltons called plantazolicin (PZN). I describe the genetic locus responsible for the biosynthesis of PZN, which is ribosomally synthesized via an amino acid precursor peptide and post-translationally modified to contain thiazoles and (methyl)oxazoles. This group of compounds, known as thiazole/oxazole-modified microcins (TOMMs), exhibit disparate biological activities and complex chemical structures. Using high-resolution mass spectrometry, chemoselective modification, genetic interruptions, and various spectroscopic tools, I report the molecular structure of PZN. PZN contains two conjugated polyazole moieties and an Nα,Nα-dimethylarginine on the amino terminus. By altering oxygenation levels during fermentation, PZN analogs were produced that bear variability in their heterocycle content, which yielded insight into the order of biosynthetic events. Extensive tailoring of PZN endows it with not only a rigid, polyheterocyclic structure, but also antibacterial activity. After screening numerous microorganisms, PZN exhibited highly selective antibiotic activity against Bacillus anthracis. This remarkably discriminatory activity rivals a previously-described B. anthracis-specific gamma (γ) phage lysis assay in distinguishing B. anthracis from other members of the Bacillus cereus group. I evaluate this unusually selective activity by measuring the RNA expression profile of PZN-treated B. anthracis, which revealed significant upregulation of genes within the cell envelope stress response. Using fluorescence microscopy, PZN localizes to distinct ~200 nm wide foci within the envelope; furthermore, like other cell envelope-acting compounds, PZN depolarizes the B. anthracis membrane. Upon selection and whole-genome sequencing of PZN-resistant mutants of B. anthracis, I implicate a relationship between the action of PZN and the phospholipid cardiolipin within the membrane. Exogenous cardiolipin increases the potency of PZN in wild type B. anthracis and promotes the incorporation of fluorescently tagged PZN in the cell envelope. I propose that PZN localizes to and exacerbates structurally compromised regions of the bacterial membrane, which ultimately results in cell lysis.","abstract_html":"Bacteria are a fruitful source of metabolites, many of which have been the scaffolds for the majority of approved antibiotic compounds. In this dissertation, I present the discovery and characterization of a bacterial natural product from Bacillus methylotrophicus FZB42, a Gram-positive, rod-shaped bacterium that stimulates plant growth. A prolific producer of secondary metabolites, FZB42 excretes a compound bearing the molecular mass of 1335 Daltons called plantazolicin (PZN). I describe the genetic locus responsible for the biosynthesis of PZN, which is ribosomally synthesized via an amino acid precursor peptide and post-translationally modified to contain thiazoles and (methyl)oxazoles. This group of compounds, known as thiazole/oxazole-modified microcins (TOMMs), exhibit disparate biological activities and complex chemical structures. Using high-resolution mass spectrometry, chemoselective modification, genetic interruptions, and various spectroscopic tools, I report the molecular structure of PZN. PZN contains two conjugated polyazole moieties and an Nα,Nα-dimethylarginine on the amino terminus. By altering oxygenation levels during fermentation, PZN analogs were produced that bear variability in their heterocycle content, which yielded insight into the order of biosynthetic events. Extensive tailoring of PZN endows it with not only a rigid, polyheterocyclic structure, but also antibacterial activity. After screening numerous microorganisms, PZN exhibited highly selective antibiotic activity against Bacillus anthracis. This remarkably discriminatory activity rivals a previously-described B. anthracis-specific gamma (γ) phage lysis assay in distinguishing B. anthracis from other members of the Bacillus cereus group. I evaluate this unusually selective activity by measuring the RNA expression profile of PZN-treated B. anthracis, which revealed significant upregulation of genes within the cell envelope stress response. Using fluorescence microscopy, PZN localizes to distinct ~200 nm wide foci within the envelope; furthermore, like other cell envelope-acting compounds, PZN depolarizes the B. anthracis membrane. Upon selection and whole-genome sequencing of PZN-resistant mutants of B. anthracis, I implicate a relationship between the action of PZN and the phospholipid cardiolipin within the membrane. Exogenous cardiolipin increases the potency of PZN in wild type B. anthracis and promotes the incorporation of fluorescently tagged PZN in the cell envelope. I propose that PZN localizes to and exacerbates structurally compromised regions of the bacterial membrane, which ultimately results in cell lysis.","abstract_has_math":false,"creators":["Molohon Hess, Katie Jo"],"institution":"University of Illinois at Urbana-Champaign","degree_name":"Ph.D.","degree_level":"Dissertation","degree_discipline":"Microbiology","degree_department":null,"school":null,"contributors":["Mitchell, Douglas A","Blanke, Steven R","Kuzminov, Andrei","Olsen, Gary J"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2016,"date_issued":"2016-03-02T20:23:46Z","date_published":"2016-03-02T20:23:46Z","updated_at":"2026-07-22T22:26:32Z","subjects":["Plantazolicin","Natural product","Bacillus anthracis","Mode of action"],"languages":["en"],"rights":["Copyright 2015 Katie Molohon Hess"],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"http://hdl.handle.net/2142/89125","outbound_label":"Handle","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Mitchell, Douglas A","Blanke, Steven R","Kuzminov, Andrei","Olsen, Gary J"]},{"key":"dc:creator","label":"Author","values":["Molohon Hess, Katie Jo"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2016-03-02T20:23:46Z","2018-03-03T10:15:31Z","2015-11-30","2015-12"]},{"key":"dc:type","label":"Dc Type","values":["text"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Microbiology"]},{"key":"thesis:degree_level","label":"Degree Level","values":["Dissertation"]},{"key":"thesis:degree_name","label":"Degree Name","values":["Ph.D."]},{"key":"thesis:institution_name","label":"Thesis Institution Name","values":["University of Illinois at Urbana-Champaign"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["Plantazolicin","Natural product","Bacillus anthracis","Mode of action"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["en"]},{"key":"dc:rights","label":"Dc Rights","values":["Copyright 2015 Katie Molohon Hess"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["http://hdl.handle.net/2142/89125"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["Bacteria are a fruitful source of metabolites, many of which have been the scaffolds for the majority of approved antibiotic compounds. In this dissertation, I present the discovery and characterization of a bacterial natural product from Bacillus methylotrophicus FZB42, a Gram-positive, rod-shaped bacterium that stimulates plant growth. A prolific producer of secondary metabolites, FZB42 excretes a compound bearing the molecular mass of 1335 Daltons called plantazolicin (PZN). I describe the genetic locus responsible for the biosynthesis of PZN, which is ribosomally synthesized via an amino acid precursor peptide and post-translationally modified to contain thiazoles and (methyl)oxazoles. This group of compounds, known as thiazole/oxazole-modified microcins (TOMMs), exhibit disparate biological activities and complex chemical structures. Using high-resolution mass spectrometry, chemoselective modification, genetic interruptions, and various spectroscopic tools, I report the molecular structure of PZN. PZN contains two conjugated polyazole moieties and an Nα,Nα-dimethylarginine on the amino terminus. By altering oxygenation levels during fermentation, PZN analogs were produced that bear variability in their heterocycle content, which yielded insight into the order of biosynthetic events. Extensive tailoring of PZN endows it with not only a rigid, polyheterocyclic structure, but also antibacterial activity. After screening numerous microorganisms, PZN exhibited highly selective antibiotic activity against Bacillus anthracis. This remarkably discriminatory activity rivals a previously-described B. anthracis-specific gamma (γ) phage lysis assay in distinguishing B. anthracis from other members of the Bacillus cereus group. I evaluate this unusually selective activity by measuring the RNA expression profile of PZN-treated B. anthracis, which revealed significant upregulation of genes within the cell envelope stress response. Using fluorescence microscopy, PZN localizes to distinct ~200 nm wide foci within the envelope; furthermore, like other cell envelope-acting compounds, PZN depolarizes the B. anthracis membrane. Upon selection and whole-genome sequencing of PZN-resistant mutants of B. anthracis, I implicate a relationship between the action of PZN and the phospholipid cardiolipin within the membrane. Exogenous cardiolipin increases the potency of PZN in wild type B. anthracis and promotes the incorporation of fluorescently tagged PZN in the cell envelope. I propose that PZN localizes to and exacerbates structurally compromised regions of the bacterial membrane, which ultimately results in cell lysis.","Submission published under a 24 month embargo labeled 'U of I Access', the embargo will last until 2017-12-01","The student, Katie Molohon Hess, accepted the attached license on 2015-11-24 at 11:22.","The student, Katie Molohon Hess, submitted this Dissertation for approval on 2015-11-24 at 11:45.","This Dissertation was approved for publication on 2015-11-30 at 15:40.","DSpace SAF Submission Ingestion Package generated from Vireo submission #8837 on 2016-03-02 at 14:06:35","Made available in DSpace on 2016-03-02T20:23:46Z (GMT). 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In this dissertation, I present the discovery and characterization of a bacterial natural product from Bacillus methylotrophicus FZB42, a Gram-positive, rod-shaped bacterium that stimulates plant growth. A prolific producer of secondary metabolites, FZB42 excretes a compound bearing the molecular mass of 1335 Daltons called plantazolicin (PZN). I describe the genetic locus responsible for the biosynthesis of PZN, which is ribosomally synthesized via an amino acid precursor peptide and post-translationally modified to contain thiazoles and (methyl)oxazoles. This group of compounds, known as thiazole/oxazole-modified microcins (TOMMs), exhibit disparate biological activities and complex chemical structures. Using high-resolution mass spectrometry, chemoselective modification, genetic interruptions, and various spectroscopic tools, I report the molecular structure of PZN. PZN contains two conjugated polyazole moieties and an Nα,Nα-dimethylarginine on the amino terminus. By altering oxygenation levels during fermentation, PZN analogs were produced that bear variability in their heterocycle content, which yielded insight into the order of biosynthetic events. Extensive tailoring of PZN endows it with not only a rigid, polyheterocyclic structure, but also antibacterial activity. After screening numerous microorganisms, PZN exhibited highly selective antibiotic activity against Bacillus anthracis. This remarkably discriminatory activity rivals a previously-described B. anthracis-specific gamma (γ) phage lysis assay in distinguishing B. anthracis from other members of the Bacillus cereus group. I evaluate this unusually selective activity by measuring the RNA expression profile of PZN-treated B. anthracis, which revealed significant upregulation of genes within the cell envelope stress response. Using fluorescence microscopy, PZN localizes to distinct ~200 nm wide foci within the envelope; furthermore, like other cell envelope-acting compounds, PZN depolarizes the B. anthracis membrane. Upon selection and whole-genome sequencing of PZN-resistant mutants of B. anthracis, I implicate a relationship between the action of PZN and the phospholipid cardiolipin within the membrane. Exogenous cardiolipin increases the potency of PZN in wild type B. anthracis and promotes the incorporation of fluorescently tagged PZN in the cell envelope. I propose that PZN localizes to and exacerbates structurally compromised regions of the bacterial membrane, which ultimately results in cell lysis.","Submission published under a 24 month embargo labeled 'U of I Access', the embargo will last until 2017-12-01","The student, Katie Molohon Hess, accepted the attached license on 2015-11-24 at 11:22.","The student, Katie Molohon Hess, submitted this Dissertation for approval on 2015-11-24 at 11:45.","This Dissertation was approved for publication on 2015-11-30 at 15:40.","DSpace SAF Submission Ingestion Package generated from Vireo submission #8837 on 2016-03-02 at 14:06:35","Made available in DSpace on 2016-03-02T20:23:46Z (GMT). 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