{"doi":"10.1111/tpj.13225","title":"Crystal structure of norcoclaurine‐6‐<i>O</i>‐methyltransferase, a key rate‐limiting step in the synthesis of benzylisoquinoline alkaloids","abstract":"<jats:title>Summary</jats:title><jats:p>Growing pharmaceutical interest in benzylisoquinoline alkaloids (<jats:styled-content style=\"fixed-case\">BIA</jats:styled-content>) coupled with their chemical complexity make metabolic engineering of microbes to create alternative platforms of production an increasingly attractive proposition. However, precise knowledge of rate‐limiting enzymes and negative feedback inhibition by end‐products of <jats:styled-content style=\"fixed-case\">BIA</jats:styled-content> metabolism is of paramount importance for this emerging field of synthetic biology. In this work we report the structural characterization of (<jats:italic>S</jats:italic>)‐norcoclaurine‐6‐<jats:italic>O</jats:italic>‐methyltransferase (6OMT), a key rate‐limiting step enzyme involved in the synthesis of reticuline, the final intermediate to be shared between the different end‐products of <jats:styled-content style=\"fixed-case\">BIA</jats:styled-content> metabolism, such as morphine, papaverine, berberine and sanguinarine. Four different crystal structures of the enzyme from <jats:italic>Thalictrum flavum</jats:italic> (Tf 6OMT) were solved: the apoenzyme, the complex with <jats:italic>S</jats:italic>‐adenosyl‐<jats:sc>l</jats:sc>‐homocysteine (<jats:styled-content style=\"fixed-case\">SAH</jats:styled-content>), the complexe with <jats:styled-content style=\"fixed-case\">SAH</jats:styled-content> and the substrate and the complex with <jats:styled-content style=\"fixed-case\">SAH</jats:styled-content> and a feedback inhibitor, sanguinarine. The Tf 6<jats:styled-content style=\"fixed-case\">OMT</jats:styled-content> structural study provides a molecular understanding of its substrate specificity, active site structure and reaction mechanism. This study also clarifies the inhibition of Tf 6<jats:styled-content style=\"fixed-case\">OMT</jats:styled-content> by previously suggested feedback inhibitors. It reveals its high and time‐dependent sensitivity toward sanguinarine.</jats:p>","journal":"The Plant Journal","year":2016,"id":673416,"datarank":0.6261580904843456,"base_score":4.174387269895637,"endowment":4.174387269895637,"self_citation_contribution":0.6261580904843456,"citation_network_contribution":0.0,"self_endowment_contribution":0.6261580904843456,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":64,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":null,"is_data_producer":false,"deposit_databanks":null,"is_oa":false,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":null,"fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1759433,"name":"Cécile Giustini","orcid":null,"position":1,"is_corresponding":false},{"id":1759434,"name":"Matthieu Graindorge","orcid":null,"position":2,"is_corresponding":false},{"id":1759435,"name":"Michel Matringe","orcid":null,"position":3,"is_corresponding":false},{"id":1759436,"name":"Renaud Dumas","orcid":null,"position":4,"is_corresponding":false},{"id":1759432,"name":"Adeline Y. Robin","orcid":null,"position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"resolved":true,"title":"Crystal structure of norcoclaurine‐6‐<i>O</i>‐methyltransferase, a key rate‐limiting step in the synthesis of benzylisoquinoline alkaloids","abstract":"<jats:title>Summary</jats:title><jats:p>Growing pharmaceutical interest in benzylisoquinoline alkaloids (<jats:styled-content style=\"fixed-case\">BIA</jats:styled-content>) coupled with their chemical complexity make metabolic engineering of microbes to create alternative platforms of production an increasingly attractive proposition. However, precise knowledge of rate‐limiting enzymes and negative feedback inhibition by end‐products of <jats:styled-content style=\"fixed-case\">BIA</jats:styled-content> metabolism is of paramount importance for this emerging field of synthetic biology. In this work we report the structural characterization of (<jats:italic>S</jats:italic>)‐norcoclaurine‐6‐<jats:italic>O</jats:italic>‐methyltransferase (6OMT), a key rate‐limiting step enzyme involved in the synthesis of reticuline, the final intermediate to be shared between the different end‐products of <jats:styled-content style=\"fixed-case\">BIA</jats:styled-content> metabolism, such as morphine, papaverine, berberine and sanguinarine. Four different crystal structures of the enzyme from <jats:italic>Thalictrum flavum</jats:italic> (Tf 6OMT) were solved: the apoenzyme, the complex with <jats:italic>S</jats:italic>‐adenosyl‐<jats:sc>l</jats:sc>‐homocysteine (<jats:styled-content style=\"fixed-case\">SAH</jats:styled-content>), the complexe with <jats:styled-content style=\"fixed-case\">SAH</jats:styled-content> and the substrate and the complex with <jats:styled-content style=\"fixed-case\">SAH</jats:styled-content> and a feedback inhibitor, sanguinarine. The Tf 6<jats:styled-content style=\"fixed-case\">OMT</jats:styled-content> structural study provides a molecular understanding of its substrate specificity, active site structure and reaction mechanism. This study also clarifies the inhibition of Tf 6<jats:styled-content style=\"fixed-case\">OMT</jats:styled-content> by previously suggested feedback inhibitors. It reveals its high and time‐dependent sensitivity toward sanguinarine.</jats:p>","is_dataset_classified":null,"base_score":4.174387269895637,"endowment":4.174387269895637,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"27232113","pmcid":null,"openalex_id":"https://openalex.org/W2400445242","authors":[],"funders":[{"funder_name":"Seventh Framework Programme","grant_id":"FP7/2007‐2013","title":null},{"funder_name":"European Commission","grant_id":"283570","title":"Transnational access and enhancement of integrated Biological Structure determination at synchrotron X-ray radiation facilities"},{"funder_name":"Institut National de la Recherche Agronomique","grant_id":"","title":null}],"total_grants":3,"fwci":5.6373,"citation_percentile":0.95648351,"influential_citations":4,"citation_trend":[{"year":2017,"count":4},{"year":2018,"count":6},{"year":2019,"count":8},{"year":2020,"count":3},{"year":2021,"count":3},{"year":2022,"count":10},{"year":2023,"count":6},{"year":2024,"count":4},{"year":2025,"count":13},{"year":2026,"count":7}],"oa_status":"closed","license":"Wiley Online Library User Agreement","oa_locations":[{"url":"https://www.onlinelibrary.wiley.com/doi/pdfdirect/10.1111/tpj.13225","host_type":"BRONZE"},{"url":"https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1111%2Ftpj.13225","host_type":"publisher"},{"url":"https://onlinelibrary.wiley.com/doi/pdf/10.1111/tpj.13225","host_type":"publisher"},{"url":"https://doi.org/10.1111/tpj.13225","host_type":"journal"},{"url":"https://pubmed.ncbi.nlm.nih.gov/27232113","host_type":"repository"},{"url":"https://hal.science/hal-01412779","host_type":"repository"},{"url":"https://dx.doi.org/10.1111/tpj.13225","host_type":""},{"url":"https://hal.science/hal-01412779v1","host_type":""},{"url":"https://doi.org/https://doi.org/10.1111/tpj.13225","host_type":""}],"fields_of_study":["Berberine and alkaloids research","Plant tissue culture and regeneration","Phytochemistry and biological activities of Ficus species","Medicine","Biology","Chemistry","0301 basic medicine","03 medical and health sciences","0303 health sciences","Benzophenanthridines","Benzylisoquinolines","Berberine","Binding Sites","Crystallography, X-Ray","Enzyme Inhibitors","Escherichia coli","Isoquinolines","Methyltransferases","Plant Proteins","Protein Conformation","Protein Multimerization","Thalictrum"],"mesh_terms":["Berberine","Binding Sites","Enzyme Inhibitors","Escherichia coli","Isoquinolines","Methyltransferases","Plant Proteins","Protein Conformation","Crystallography, X-Ray","Thalictrum","Benzylisoquinolines","Benzophenanthridines","Protein Multimerization"],"keywords":["Benzylisoquinoline","Sanguinarine","Biology","Papaverine","Berberine","Alkaloid","Enzyme","Stereochemistry","Biochemistry","Biosynthesis","Chemistry","Botany","Structure","secondary metabolism","Benzylisoquinoline Alkaloids","Thalictrum Flavum","Norcoclaurine-6-o-methyltransferase","570","Protein Conformation","[SDV]Life Sciences [q-bio]","[SDV.BBM]Life Sciences [q-bio]/Biochemistry","Antimicrobial agent","Norcoclaurine - 6-0-methyltransferase","Crystallography, X-Ray","Benzylisoquinolines","Thalictrum","[SDV.BBM] Life Sciences [q-bio]/Biochemistry, Molecular Biology","Escherichia coli","Enzyme Inhibitors","Molecular Biology","Plant Proteins","Benzophenanthridines","Alkaloid biosynthesis","Binding Sites","Crystal structure","Plant","Methyltransferases","540","Isoquinolines","[SDV] Life Sciences [q-bio]","Protein Multimerization","Ranunculaceae"],"sdg_mappings":[],"linked_datasets":[],"clinical_trials":[],"software_tools":[],"database_accessions":[{"name":"pdb"}],"source":"live","citation_network_status":"fetched"},"created_at":"2026-08-16T13:23:10.583998Z","pmid":null,"pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}