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Its root is an important traditional folk medicine, which is called Diburong in China. Diburong is rich in benzylisoquinoline alkaloids (BIAs), including cepharanthine, which has been demonstrated to exhibit significant anti-inflammatory, antiviral, antineoplastic, and anti-SARS-CoV-2 activities, as well as raising leukocytes. Cepharanthine is composed of (R)- and (S)-1-benzylisoquinoline alkaloid (1-BIA). (S)-norcoclaurine-6-O-methyltransferase (6OMT) is a rate-limiting enzyme in BIA biosynthesis. However, its role in the cepharanthine biosynthetic pathway, particularly with the (R) stereoisomer substrate, remains largely unexplored. This study aimed to identify Se6OMTs involved in the cepharanthine biosynthetic pathway and elucidate the O-methyltransferases (OMTs) responsible for the production of (R)- and (S)-stereoisomer BIAs. Methods: In this study, three OMTs were cloned from S. epigaea and functionally characterized using nine 1-BIAs of (R)- and (S)-configurations as substrates. Results: Two O-methyltransferases, Se6OMT1 and Se6OMT3, showed efficient catalytic activity at the C6 position of both (R)- and (S)-norcoclaurine. Furthermore, Se6OMT3 demonstrated high catalytic activity at the C7 and C4\u2032 positions of other (R)- and (S)-configuration 1-BIAs, which resulted in the generation of multiple products. Conclusions: This study focused on 6OMT enzymes in S. epigaea, identifying Se6OMTs involved in the cepharanthine biosynthetic pathway, determining the OMTs involved in the production of (R)- and (S)-stereoisomer BIAs. This research provides valuable insights into the substrate promiscuity of Se6OMTs on (R)- and (S)-configured 1-BIAs in S. epigaea and highlights the genetic components necessary for the metabolic engineering and synthetic biology approaches to cepharanthine production.<\/jats:p>","DOI":"10.3390\/metabo15020092","type":"journal-article","created":{"date-parts":[[2025,2,4]],"date-time":"2025-02-04T05:13:43Z","timestamp":1738646023000},"page":"92","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Identification and Characterization of Two Se6OMTs from Stephania epigaea Offer Novel Insights into the Biosynthetic Pathway of Cepharanthine"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0009-0004-9261-6159","authenticated-orcid":false,"given":"Jingyi","family":"Gan","sequence":"first","affiliation":[{"name":"College of Traditional Chinese Medicine, Yunnan University of Chinese Medicine, Kunming 650500, China"},{"name":"State Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China"},{"name":"Key Laboratory of Yunnan Provincial Department of Education on Substance Benchmark Research of Ethnic Medicines, Yunnan University of Chinese Medicine, Kunming 650500, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-6333-7137","authenticated-orcid":false,"given":"Wenlong","family":"Shi","sequence":"additional","affiliation":[{"name":"State Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2148-7917","authenticated-orcid":false,"given":"Qishuang","family":"Li","sequence":"additional","affiliation":[{"name":"State Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xinyi","family":"Li","sequence":"additional","affiliation":[{"name":"State Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xingyu","family":"Zhao","sequence":"additional","affiliation":[{"name":"College of Traditional Chinese Medicine, Yunnan University of Chinese Medicine, Kunming 650500, China"},{"name":"State Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Junhao","family":"Tang","sequence":"additional","affiliation":[{"name":"College of Traditional Chinese Medicine, Yunnan University of Chinese Medicine, Kunming 650500, China"},{"name":"State Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ying","family":"Ma","sequence":"additional","affiliation":[{"name":"State Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9386-7986","authenticated-orcid":false,"given":"Jian","family":"Wang","sequence":"additional","affiliation":[{"name":"State Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shukun","family":"Gong","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Southwestern Chinese Medicine Resources, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5961-3168","authenticated-orcid":false,"given":"Xiaohui","family":"Ma","sequence":"additional","affiliation":[{"name":"College of Traditional Chinese Medicine, Yunnan University of Chinese Medicine, Kunming 650500, China"},{"name":"Key Laboratory of Yunnan Provincial Department of Education on Substance Benchmark Research of Ethnic Medicines, Yunnan University of Chinese Medicine, Kunming 650500, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Juan","family":"Guo","sequence":"additional","affiliation":[{"name":"State Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China"},{"name":"Dalian Institute of Marine Traditional Chinese Medicine, Dalian 116000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,2,3]]},"reference":[{"key":"ref_1","unstructured":"Wu, Z., Raven, P.H., and Deyuan, H. (1996). Missouri Botanical Garden. Flora of China, Science Press."},{"key":"ref_2","unstructured":"Administration, F.P.F.a.D (2012). Fujian Province Processing Standards for Traditional Chinese Medicine Pieces, Fujian Science and Technology Press."},{"key":"ref_3","unstructured":"Administration, Y.P.F.a.D (2005). Yunnan Province Traditional Chinese Medicine Standards 2005 Edition, Yunnan Fine Arts Publishing House."},{"key":"ref_4","first-page":"89","article-title":"Progress in Chemical Constituents, Pharmacology and Fermentation Modification of Radix Stephaniae epigaeae","volume":"52","author":"Li","year":"2023","journal-title":"Shandong Chem. Ind."},{"key":"ref_5","first-page":"3641586","article-title":"Identification of Bioactive Components of Stephania epigaea Lo and Their Potential Therapeutic Targets by UPLC-MS\/MS and Network Pharmacology","volume":"2022","author":"Li","year":"2022","journal-title":"Evid. Based Complement. Alternat. Med."},{"key":"ref_6","unstructured":"Yang, L. (2019). Research on the Chemical Constituents and Biological Activities of Sinomenine and Stephania epigaea, Yunnan University."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1509","DOI":"10.1007\/s43440-020-00132-z","article-title":"Cepharanthine: A review of the antiviral potential of a Japanese-approved alopecia drug in COVID-19","volume":"72","author":"Rogosnitzky","year":"2020","journal-title":"Pharmacol. Rep."},{"key":"ref_8","first-page":"955","article-title":"Progress in the study of antiviral activity of cepharanthine against SARS-CoV-2","volume":"42","author":"Fan","year":"2022","journal-title":"J. South. Med. Univ."},{"key":"ref_9","first-page":"118","article-title":"Improvement on the Determination Method of Cepharanthine Tablets","volume":"03","author":"Xiao","year":"2005","journal-title":"Chin. Tradit. Pat. Med."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1051","DOI":"10.1097\/CM9.0000000000000797","article-title":"Repurposing of clinically approved drugs for treatment of coronavirus disease 2019 in a 2019-novel coronavirus-related coronavirus model","volume":"133","author":"Fan","year":"2020","journal-title":"Chin. Med. J."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1537","DOI":"10.1038\/s41467-024-45690-5","article-title":"Cepharanthine analogs mining and genomes of Stephania accelerate anti-coronavirus drug discovery","volume":"15","author":"Leng","year":"2024","journal-title":"Nat. Commun."},{"key":"ref_12","first-page":"61","article-title":"Determination of Cepharanthine and Cycleanine in Stephania epigaea H.S.Lo by RP-HPLC","volume":"24","author":"Li","year":"2010","journal-title":"Res. Pract. Chin. Med. Mod."},{"key":"ref_13","first-page":"2079","article-title":"Analysis for biosynthetic pathways of active constituents in medicinal plants and its availability","volume":"32","author":"Ma","year":"2017","journal-title":"Chin. J. Tradit. Chin. Med."},{"key":"ref_14","first-page":"121","article-title":"Biosynthesis of the Bisbenzylisoquinoline Alkaloid Cocsulin","volume":"1","author":"Bhakuni","year":"1978","journal-title":"J. Chem. Soc."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"2347","DOI":"10.1016\/S0031-9422(00)91024-0","article-title":"Biosynthesis of the bisbenzylisoquinoline alkaloid, tetrandrine","volume":"19","author":"Bhakuni","year":"1980","journal-title":"Phytochemistry"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"2071","DOI":"10.1073\/pnas.92.6.2071","article-title":"Molecular cloning and heterologous expression of a cDNA encoding berbamunine synthase, a C--O phenol-coupling cytochrome P450 from the higher plant Berberis stolonifera","volume":"92","author":"Kraus","year":"1995","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"162","DOI":"10.1016\/j.ymben.2023.03.010","article-title":"Pathway elucidation and microbial synthesis of proaporphine and bis-benzylisoquinoline alkaloids from sacred lotus (Nelumbo nucifera)","volume":"77","author":"Pyne","year":"2023","journal-title":"Metab. Eng."},{"key":"ref_18","first-page":"716","article-title":"Research progress and challenges in microbial synthesis of benzylisoquinoline alkaloids","volume":"2","author":"Lin","year":"2021","journal-title":"Synth. Biol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"641","DOI":"10.1111\/tpj.13225","article-title":"Crystal structure of norcoclaurine-6-O-methyltransferase, a key rate-limiting step in the synthesis of benzylisoquinoline alkaloids","volume":"87","author":"Robin","year":"2016","journal-title":"Plant J."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"153181","DOI":"10.1016\/j.jplph.2020.153181","article-title":"Characterization of O-methyltransferases involved in the biosynthesis of tetrandrine in Stephania tetrandra","volume":"250","author":"Li","year":"2020","journal-title":"J. Plant Physiol."},{"key":"ref_21","unstructured":"Romeo, J.T. (2003). Chapter two Structural, functional, and evolutionary basis for methylation of plant small molecules. Recent Advances in Phytochemistry, Elsevier."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"252","DOI":"10.1093\/pcp\/pcl062","article-title":"Overexpression of Coptis japonica norcoclaurine 6-O-methyltransferase overcomes the rate-limiting step in Benzylisoquinoline alkaloid biosynthesis in cultured Eschscholzia californica","volume":"48","author":"Inui","year":"2007","journal-title":"Plant Cell Physiol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"23398","DOI":"10.1074\/jbc.M002439200","article-title":"Molecular Characterization of the S-Adenosyl-l-methionine:3\u2032-Hydroxy-N-methylcoclaurine 4\u2032-O-Methyltransferase Involved in Isoquinoline Alkaloid Biosynthesis in Coptis japonica","volume":"275","author":"Morishige","year":"2000","journal-title":"J. Biol. Chem."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Li, K., Chen, X., Zhang, J., Wang, C., Xu, Q., Hu, J., Kai, G., and Feng, Y. (2022). Transcriptome Analysis of Stephania tetrandra and Characterization of Norcoclaurine-6-O-Methyltransferase Involved in Benzylisoquinoline Alkaloid Biosynthesis. Front. Plant Sci., 13.","DOI":"10.3389\/fpls.2022.874583"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1038\/75556","article-title":"Gene ontology: Tool for the unification of biology. The Gene Ontology Consortium","volume":"25","author":"Ashburner","year":"2000","journal-title":"Nat. Genet."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"D277","DOI":"10.1093\/nar\/gkh063","article-title":"The KEGG resource for deciphering the genome","volume":"32","author":"Kanehisa","year":"2004","journal-title":"Nucleic Acids Res."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"D115","DOI":"10.1093\/nar\/gkh131","article-title":"UniProt: The Universal Protein knowledgebase","volume":"32","author":"Apweiler","year":"2004","journal-title":"Nucleic Acids Res."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Tatusov, R.L., Fedorova, N.D., Jackson, J.D., Jacobs, A.R., Kiryutin, B., Koonin, E.V., Krylov, D.M., Mazumder, R., Mekhedov, S.L., and Nikolskaya, A.N. (2003). The COG database: An updated version includes eukaryotes. BMC Bioinform., 4.","DOI":"10.1186\/1471-2105-4-41"},{"key":"ref_29","first-page":"71","article-title":"Integrated nr Database in Protein Annotation System and Its Localization","volume":"32","author":"Deng","year":"2006","journal-title":"Comput. Eng."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"D222","DOI":"10.1093\/nar\/gkt1223","article-title":"Pfam: The protein families database","volume":"42","author":"Finn","year":"2014","journal-title":"Nucleic Acids Res."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1703","DOI":"10.1111\/jipb.13724","article-title":"Identification of the cytochrome P450s responsible for the biosynthesis of two types of aporphine alkaloids and their de novo biosynthesis in yeast","volume":"66","author":"Li","year":"2024","journal-title":"J. Integr. Plant Biol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1238","DOI":"10.1039\/c2np20029e","article-title":"Architectures, mechanisms and molecular evolution of natural product methyltransferases","volume":"29","author":"Liscombe","year":"2012","journal-title":"Nat. Prod. Rep."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"547","DOI":"10.1111\/j.1365-313X.2006.02801.x","article-title":"The role of phloem sieve elements and laticifers in the biosynthesis and accumulation of alkaloids in opium poppy","volume":"47","author":"Samanani","year":"2006","journal-title":"Plant J."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"He, S.-M., Liang, Y.-L., Cong, K., Chen, G., Zhao, X., Zhao, Q.-M., Zhang, J.-J., Wang, X., Dong, Y., and Yang, J.-L. (2018). Identification and Characterization of Genes Involved in Benzylisoquinoline Alkaloid Biosynthesis in Coptis Species. Front. Plant Sci., 9.","DOI":"10.3389\/fpls.2018.00731"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"e2112520118","DOI":"10.1073\/pnas.2112520118","article-title":"Complete biosynthesis of the bisbenzylisoquinoline alkaloids guattegaumerine and berbamunine in yeast","volume":"118","author":"Payne","year":"2021","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"uhac152","DOI":"10.1093\/hr\/uhac152","article-title":"Catalytic promiscuity of O-methyltransferases from Corydalis yanhusuo leading to the structural diversity of benzylisoquinoline alkaloids","volume":"9","author":"Bu","year":"2022","journal-title":"Hortic. Res."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"735","DOI":"10.1146\/annurev.arplant.59.032607.092730","article-title":"Alkaloid biosynthesis: Metabolism and trafficking","volume":"59","author":"Ziegler","year":"2008","journal-title":"Annu. Rev. Plant. Biol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"102367","DOI":"10.1016\/j.isci.2021.102367","article-title":"Potential anti-COVID-19 agents, cepharanthine and nelfinavir, and their usage for combination treatment","volume":"24","author":"Ohashi","year":"2021","journal-title":"iScience"},{"key":"ref_39","first-page":"1869","article-title":"Application of Methyltransferase in Microbial Synthesis of Natural Products","volume":"37","author":"Zhang","year":"2021","journal-title":"J. Bioeng."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1127","DOI":"10.1104\/pp.15.01240","article-title":"Isolation and Characterization of O-methyltransferases Involved in the Biosynthesis of Glaucine in Glaucium flavum","volume":"169","author":"Chang","year":"2015","journal-title":"Plant Physiol."},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Morris, J.S., and Facchini, P.J. (2019). Molecular Origins of Functional Diversity in Benzylisoquinoline Alkaloid Methyltransferases. Front. Plant Sci., 10.","DOI":"10.3389\/fpls.2019.01058"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"4497","DOI":"10.1021\/acscatal.9b05417","article-title":"Structure-Guided Engineering of a Scoulerine 9-O-Methyltransferase Enables the Biosynthesis of Tetrahydropalmatrubine and Tetrahydropalmatine in Yeast","volume":"10","author":"Valentic","year":"2020","journal-title":"ACS Catal."}],"container-title":["Metabolites"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2218-1989\/15\/2\/92\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T16:26:05Z","timestamp":1760027165000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2218-1989\/15\/2\/92"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,2,3]]},"references-count":42,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2025,2]]}},"alternative-id":["metabo15020092"],"URL":"https:\/\/doi.org\/10.3390\/metabo15020092","relation":{},"ISSN":["2218-1989"],"issn-type":[{"value":"2218-1989","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,2,3]]}}}