{"id":{"repo_id":"uwo","oai_identifier":"oai:uwo.scholaris.ca:20.500.14721/39220"},"canonical_url":"https://search.dev.ndltd.org/etd/uwo/oai:uwo.scholaris.ca:20.500.14721/39220","repository":{"repo_id":"uwo","name":"Western University","base_url":"https://uwo.scholaris.ca/server/oai/request"},"display":{"title":"Biosynthesis of tetrahydroisoquinoline scaffolds in yeast","abstract":"In previous work, Saccharomyces cerevisiae was used to produce the tetrahydroisoquinoline alkaloid precursor (S)-norcoclaurine via a combination of yeast Ehrlich pathway metabolism of ʟ-tyrosine and expression of norcoclaurine synthase with a dopamine production pathway. Building on this, I developed a retrosynthetic framework to produce phenylisoquinoline, phenethylisoquinoline, and novel phenylpropylisoquinoline scaffolds. By feeding engineered yeast nonstandard amino acids, which can be metabolized by the yeast Ehrlich pathway, scaffolds for tetrahydroisoquinoline alkaloids were detected and their relative abundance quantified using mass spectrometry. Engineering many strains with different norcoclaurine synthase orthologs and feeding them different nonstandard amino acids led to the identification of orthologs which are uniquely adept at producing each class of tetrahydroisoquinoline. By selective knockout of oxidoreductase improvements in titers of phenylpropylisoquinoline were achieved. Biosynthesis of these scaffolds, the committed step in yeasts provides a crucial step in the de novo biosynthesis of tetrahydroisoquinoline compounds, providing an alternative means production.","abstract_html":"In previous work, Saccharomyces cerevisiae was used to produce the tetrahydroisoquinoline alkaloid precursor (S)-norcoclaurine via a combination of yeast Ehrlich pathway metabolism of ʟ-tyrosine and expression of norcoclaurine synthase with a dopamine production pathway. Building on this, I developed a retrosynthetic framework to produce phenylisoquinoline, phenethylisoquinoline, and novel phenylpropylisoquinoline scaffolds. By feeding engineered yeast nonstandard amino acids, which can be metabolized by the yeast Ehrlich pathway, scaffolds for tetrahydroisoquinoline alkaloids were detected and their relative abundance quantified using mass spectrometry. Engineering many strains with different norcoclaurine synthase orthologs and feeding them different nonstandard amino acids led to the identification of orthologs which are uniquely adept at producing each class of tetrahydroisoquinoline. By selective knockout of oxidoreductase improvements in titers of phenylpropylisoquinoline were achieved. Biosynthesis of these scaffolds, the committed step in yeasts provides a crucial step in the de novo biosynthesis of tetrahydroisoquinoline compounds, providing an alternative means production.","abstract_has_math":false,"creators":["Kowalski, Devin"],"institution":"The University of Western Ontario","degree_name":"M Sc","degree_level":null,"degree_discipline":"Biology","degree_department":null,"school":null,"contributors":[],"advisors":["Michael, Pyne"],"committee_chairs":[],"committee_members":[],"year":2025,"date_issued":"2025-11-04","date_published":"2025-11-04","updated_at":"2026-07-27T21:55:59Z","subjects":["norcoclaurine synthase","synthetic biology","metabolic engineering","phenylisoquinoline","phenethylisoquinoline","phenylpropylisoquinoline"],"languages":["en"],"rights":["Attribution-NonCommercial-NoDerivatives 4.0 International"],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"https://hdl.handle.net/20.500.14721/39220","outbound_label":"Handle","outbound_source":"dc:identifier.uri"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor.advisor","label":"Advisor","values":["Michael, Pyne"]},{"key":"dc:creator","label":"Author","values":["Kowalski, Devin"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.accessioned","label":"Dc Date Accessioned","values":["2025-12-10T18:08:43Z"]},{"key":"dc:date.issued","label":"Date","values":["2025-11-04"]},{"key":"dc:publisher","label":"Institution","values":["The University of Western Ontario"]},{"key":"dc:type","label":"Dc Type","values":["thesis"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Biology"]},{"key":"thesis:degree_name","label":"Degree Name","values":["M Sc"]},{"key":"thesis:institution_name","label":"Thesis Institution Name","values":["The University of Western Ontario"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["norcoclaurine synthase","synthetic biology","metabolic engineering","phenylisoquinoline","phenethylisoquinoline","phenylpropylisoquinoline"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language.iso","label":"Language (ISO)","values":["en"]},{"key":"dc:rights","label":"Dc Rights","values":["Attribution-NonCommercial-NoDerivatives 4.0 International"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier.uri","label":"Identifier URI","values":["https://hdl.handle.net/20.500.14721/39220"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["In previous work, Saccharomyces cerevisiae was used to produce the tetrahydroisoquinoline alkaloid precursor (S)-norcoclaurine via a combination of yeast Ehrlich pathway metabolism of ʟ-tyrosine and expression of norcoclaurine synthase with a dopamine production pathway. Building on this, I developed a retrosynthetic framework to produce phenylisoquinoline, phenethylisoquinoline, and novel phenylpropylisoquinoline scaffolds. By feeding engineered yeast nonstandard amino acids, which can be metabolized by the yeast Ehrlich pathway, scaffolds for tetrahydroisoquinoline alkaloids were detected and their relative abundance quantified using mass spectrometry. Engineering many strains with different norcoclaurine synthase orthologs and feeding them different nonstandard amino acids led to the identification of orthologs which are uniquely adept at producing each class of tetrahydroisoquinoline. By selective knockout of oxidoreductase improvements in titers of phenylpropylisoquinoline were achieved. Biosynthesis of these scaffolds, the committed step in yeasts provides a crucial step in the de novo biosynthesis of tetrahydroisoquinoline compounds, providing an alternative means production."]},{"key":"dc:title","label":"Title","values":["Biosynthesis of tetrahydroisoquinoline scaffolds in yeast"]}]}],"canonical_facts":{"dc:contributor.advisor":["Michael, Pyne"],"dc:creator":["Kowalski, Devin"],"dc:date.accessioned":["2025-12-10T18:08:43Z"],"dc:date.issued":["2025-11-04"],"dc:description.abstract":["In previous work, Saccharomyces cerevisiae was used to produce the tetrahydroisoquinoline alkaloid precursor (S)-norcoclaurine via a combination of yeast Ehrlich pathway metabolism of ʟ-tyrosine and expression of norcoclaurine synthase with a dopamine production pathway. Building on this, I developed a retrosynthetic framework to produce phenylisoquinoline, phenethylisoquinoline, and novel phenylpropylisoquinoline scaffolds. By feeding engineered yeast nonstandard amino acids, which can be metabolized by the yeast Ehrlich pathway, scaffolds for tetrahydroisoquinoline alkaloids were detected and their relative abundance quantified using mass spectrometry. Engineering many strains with different norcoclaurine synthase orthologs and feeding them different nonstandard amino acids led to the identification of orthologs which are uniquely adept at producing each class of tetrahydroisoquinoline. By selective knockout of oxidoreductase improvements in titers of phenylpropylisoquinoline were achieved. Biosynthesis of these scaffolds, the committed step in yeasts provides a crucial step in the de novo biosynthesis of tetrahydroisoquinoline compounds, providing an alternative means production."],"dc:identifier.uri":["https://hdl.handle.net/20.500.14721/39220"],"dc:language.iso":["en"],"dc:publisher":["The University of Western Ontario"],"dc:rights":["Attribution-NonCommercial-NoDerivatives 4.0 International"],"dc:subject":["norcoclaurine synthase","synthetic biology","metabolic engineering","phenylisoquinoline","phenethylisoquinoline","phenylpropylisoquinoline"],"dc:title":["Biosynthesis of tetrahydroisoquinoline scaffolds in yeast"],"dc:type":["thesis"],"thesis:degree_discipline":["Biology"],"thesis:degree_name":["M Sc"],"thesis:institution_name":["The University of Western Ontario"]},"updated_at":"2026-07-27T21:55:59Z"}