{"id":{"repo_id":"uiuc","oai_identifier":"oai:www.ideals.illinois.edu:2142/105122"},"canonical_url":"https://search.dev.ndltd.org/etd/uiuc/oai:www.ideals.illinois.edu:2142/105122","repository":{"repo_id":"uiuc","name":"University of Illinois - Urbana-Champaign","base_url":"https://www.ideals.illinois.edu/oai-pmh"},"display":{"title":"Biochemical characterization of enzymes involved in the biosynthesis of the thiopeptide thiomuracin","abstract":"Clinically significant antibiotic resistance has evolved against virtually every antibiotic currently deployed, therefore it is critical for human health to discover new antibacterial agents with novel modes of action. Thiopeptides are a family of ribosomally synthesized and post-translationally modified peptides (RiPPs) that have outstanding biological profile, including high potency against a variety of antibiotic resistant pathogenic strains. Their new modes of action have attracted many efforts to develop chemical syntheses, and to study their biological function and biosynthetic origin. However, the complex architecture and poor physicochemical properties have plagued this otherwise-promising class of antibiotics. To address these challenges, this dissertation used the thiopeptide thiomuracin as a model to establish the reconstitution of the biosynthesis of thiopeptide in vitro, and to characterize the activity of the highly unusual modification enzymes involved in thiopeptide biosynthetic pathway. This work will guide future efforts to improve the properties of thiopeptide natural products. The biosynthesis of the thiopeptide thiomuracin is a well-orchestrated process involving a multitude of post-translational modifications. Chapter 2 presents the first in vitro biosynthesis of the core scaffold of thiopeptide thiomuracin, while Chapter 3 will provide insights into the substrate specificities, directionality, and timing of catalysis by six proteins involved in the formation of this core scaffold of thiopeptides. Characterization and mechanistic investigation of TbtI, a radical S-adenosyl-L-methionine thiazole C-methyltransferase involved in the tailoring modification of thiomuracin, are presented in Chapter 4. Lastly, Chapter 5 discusses the characterization of an ATP and tRNA dependent peptidyl transferase involved in a pathway where a ribosomally synthesized small peptide serves as a catalytic scaffold on which a small-molecule is biosynthesized in Pseudomonas syringae.","abstract_html":"Clinically significant antibiotic resistance has evolved against virtually every antibiotic currently deployed, therefore it is critical for human health to discover new antibacterial agents with novel modes of action. Thiopeptides are a family of ribosomally synthesized and post-translationally modified peptides (RiPPs) that have outstanding biological profile, including high potency against a variety of antibiotic resistant pathogenic strains. Their new modes of action have attracted many efforts to develop chemical syntheses, and to study their biological function and biosynthetic origin. However, the complex architecture and poor physicochemical properties have plagued this otherwise-promising class of antibiotics. To address these challenges, this dissertation used the thiopeptide thiomuracin as a model to establish the reconstitution of the biosynthesis of thiopeptide in vitro, and to characterize the activity of the highly unusual modification enzymes involved in thiopeptide biosynthetic pathway. This work will guide future efforts to improve the properties of thiopeptide natural products. The biosynthesis of the thiopeptide thiomuracin is a well-orchestrated process involving a multitude of post-translational modifications. Chapter 2 presents the first in vitro biosynthesis of the core scaffold of thiopeptide thiomuracin, while Chapter 3 will provide insights into the substrate specificities, directionality, and timing of catalysis by six proteins involved in the formation of this core scaffold of thiopeptides. Characterization and mechanistic investigation of TbtI, a radical S-adenosyl-L-methionine thiazole C-methyltransferase involved in the tailoring modification of thiomuracin, are presented in Chapter 4. Lastly, Chapter 5 discusses the characterization of an ATP and tRNA dependent peptidyl transferase involved in a pathway where a ribosomally synthesized small peptide serves as a catalytic scaffold on which a small-molecule is biosynthesized in Pseudomonas syringae.","abstract_has_math":false,"creators":["Zhang, Zhengan"],"institution":"University of Illinois at Urbana-Champaign","degree_name":"Ph.D.","degree_level":"Dissertation","degree_discipline":"Chemistry","degree_department":null,"school":null,"contributors":["van der Donk, Wilfred A.","Mitchell, Douglas A.","Nair, Satish K.","Gerlt, John A."],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2019,"date_issued":"2019-08-23T20:44:22Z","date_published":"2019-08-23T20:44:22Z","updated_at":"2026-07-22T22:24:44Z","subjects":["None"],"languages":["en"],"rights":["Copyright 2019 Zhengan Zhang"],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"http://hdl.handle.net/2142/105122","outbound_label":"Handle","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["van der Donk, Wilfred A.","Mitchell, Douglas A.","Nair, Satish K.","Gerlt, John A."]},{"key":"dc:creator","label":"Author","values":["Zhang, Zhengan"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2019-08-23T20:44:22Z","2021-08-24T09:15:28Z","2018-12-20","2019-05"]},{"key":"dc:type","label":"Dc Type","values":["text"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Chemistry"]},{"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":["None"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["en"]},{"key":"dc:rights","label":"Dc Rights","values":["Copyright 2019 Zhengan Zhang"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["http://hdl.handle.net/2142/105122"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["Clinically significant antibiotic resistance has evolved against virtually every antibiotic currently deployed, therefore it is critical for human health to discover new antibacterial agents with novel modes of action. Thiopeptides are a family of ribosomally synthesized and post-translationally modified peptides (RiPPs) that have outstanding biological profile, including high potency against a variety of antibiotic resistant pathogenic strains. Their new modes of action have attracted many efforts to develop chemical syntheses, and to study their biological function and biosynthetic origin. However, the complex architecture and poor physicochemical properties have plagued this otherwise-promising class of antibiotics. To address these challenges, this dissertation used the thiopeptide thiomuracin as a model to establish the reconstitution of the biosynthesis of thiopeptide in vitro, and to characterize the activity of the highly unusual modification enzymes involved in thiopeptide biosynthetic pathway. This work will guide future efforts to improve the properties of thiopeptide natural products. The biosynthesis of the thiopeptide thiomuracin is a well-orchestrated process involving a multitude of post-translational modifications. Chapter 2 presents the first in vitro biosynthesis of the core scaffold of thiopeptide thiomuracin, while Chapter 3 will provide insights into the substrate specificities, directionality, and timing of catalysis by six proteins involved in the formation of this core scaffold of thiopeptides. Characterization and mechanistic investigation of TbtI, a radical S-adenosyl-L-methionine thiazole C-methyltransferase involved in the tailoring modification of thiomuracin, are presented in Chapter 4. Lastly, Chapter 5 discusses the characterization of an ATP and tRNA dependent peptidyl transferase involved in a pathway where a ribosomally synthesized small peptide serves as a catalytic scaffold on which a small-molecule is biosynthesized in Pseudomonas syringae.","Submission published under a 24 month embargo labeled 'Closed Access', the embargo will last until 2021-05-01","The student, Zhengan Zhang, accepted the attached license on 2018-12-18 at 12:03.","The student, Zhengan Zhang, submitted this Dissertation for approval on 2018-12-18 at 12:15.","This Dissertation was approved for publication on 2018-12-20 at 08:42.","DSpace SAF Submission Ingestion Package generated from Vireo submission #13341 on 2019-08-22 at 16:18:20","Made available in DSpace on 2019-08-23T20:44:22Z (GMT). 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Thiopeptides are a family of ribosomally synthesized and post-translationally modified peptides (RiPPs) that have outstanding biological profile, including high potency against a variety of antibiotic resistant pathogenic strains. Their new modes of action have attracted many efforts to develop chemical syntheses, and to study their biological function and biosynthetic origin. However, the complex architecture and poor physicochemical properties have plagued this otherwise-promising class of antibiotics. To address these challenges, this dissertation used the thiopeptide thiomuracin as a model to establish the reconstitution of the biosynthesis of thiopeptide in vitro, and to characterize the activity of the highly unusual modification enzymes involved in thiopeptide biosynthetic pathway. This work will guide future efforts to improve the properties of thiopeptide natural products. The biosynthesis of the thiopeptide thiomuracin is a well-orchestrated process involving a multitude of post-translational modifications. Chapter 2 presents the first in vitro biosynthesis of the core scaffold of thiopeptide thiomuracin, while Chapter 3 will provide insights into the substrate specificities, directionality, and timing of catalysis by six proteins involved in the formation of this core scaffold of thiopeptides. Characterization and mechanistic investigation of TbtI, a radical S-adenosyl-L-methionine thiazole C-methyltransferase involved in the tailoring modification of thiomuracin, are presented in Chapter 4. Lastly, Chapter 5 discusses the characterization of an ATP and tRNA dependent peptidyl transferase involved in a pathway where a ribosomally synthesized small peptide serves as a catalytic scaffold on which a small-molecule is biosynthesized in Pseudomonas syringae.","Submission published under a 24 month embargo labeled 'Closed Access', the embargo will last until 2021-05-01","The student, Zhengan Zhang, accepted the attached license on 2018-12-18 at 12:03.","The student, Zhengan Zhang, submitted this Dissertation for approval on 2018-12-18 at 12:15.","This Dissertation was approved for publication on 2018-12-20 at 08:42.","DSpace SAF Submission Ingestion Package generated from Vireo submission #13341 on 2019-08-22 at 16:18:20","Made available in DSpace on 2019-08-23T20:44:22Z (GMT). No. of bitstreams: 6 ZHANG-DISSERTATION-2019.pdf: 7906170 bytes, checksum: 6a3eec02d2abe0cc0a34c59616cd11bb (MD5) Copyright Biosynthetic timing paper.pdf: 73031 bytes, checksum: a01a797352f74408268bd795725746fd (MD5) Copyright Mechanism of a Class C Radical SAM paper.pdf: 75896 bytes, checksum: de4268c4d94e867398ed574ae076406a (MD5) Copyright Reconstitution and Substrate Specificity of the Radical SAM paper.pdf: 72904 bytes, checksum: 691241b661c9f4703d0cf3850b52c7ca (MD5) LICENSE.txt: 4210 bytes, checksum: a85eceb251365a56ea6b0f6e5a701975 (MD5) RightsLink Printable License figure 1.1.pdf: 110528 bytes, checksum: 438ae2e4d01bb039e13fa88ba89f5855 (MD5) Previous issue date: 2018-12-20","Embargo set by: Seth Robbins for item 112241 Lift date: 2021-08-23T20:44:50Z Reason: Author requested closed access (OA after 2yrs) in Vireo ETD system","Embargo set by: Seth Robbins for item 112241 Lift date: 2021-08-23T20:46:41Z Reason: Author requested closed access (OA after 2yrs) in Vireo ETD system","Embargo set by: Seth Robbins for item 112241 Lift date: 2021-08-23T20:47:38Z Reason: Author requested closed access (OA after 2yrs) in Vireo ETD system","Embargo set by: Seth Robbins for item 112241 Lift date: 2021-08-23T20:48:32Z Reason: Author requested closed access (OA after 2yrs) in Vireo ETD system","Limited Restriction Lifted for Item 112241 on 2021-08-24T09:15:28Z."],"dc:format":["application/pdf"],"dc:identifier":["http://hdl.handle.net/2142/105122"],"dc:language":["en"],"dc:rights":["Copyright 2019 Zhengan Zhang"],"dc:subject":["None"],"dc:title":["Biochemical characterization of enzymes involved in the biosynthesis of the thiopeptide thiomuracin"],"dc:type":["text"],"thesis:degree_discipline":["Chemistry"],"thesis:degree_level":["Dissertation"],"thesis:degree_name":["Ph.D."],"thesis:institution_name":["University of Illinois at Urbana-Champaign"]},"updated_at":"2026-07-22T22:24:44Z"}