{
  "abstract": "Astragalus membranaceus , a widely used medicinal and edible herb, contains major active isoflavonoid components such as formononetin, calycosin, and their glycosides, which exhibit a broad spectrum of pharmacological effects including anti-inflammatory, immunomodulatory, and cardioprotective activities. In this study, we identified three key enzymes: isoflavone synthase (AmIFS), O -methyltransferase (AmOMT2), and isoflavone 3′-hydroxylase (AmI3′H), elucidating the critical steps in calycosin biosynthesis and revealing a novel synthetic pathway. The research showed that AmIFS efficiently catalyzed the conversion of liquiritigenin and naringenin into the isoflavone backbone. The multifunctional O -methyltransferase AmOMT2 preferentially catalyzed the 4′-O-methylation of liquiritigenin, and the resulting product was subsequently converted by AmIFS into 2,7-dihydroxy-4′-methoxyisoflavanone, thereby broadening substrate versatility of AmIFS and revealing a new biosynthetic route. Meanwhile, AmI3′H completed the biosynthesis by catalyzing the final 3′-hydroxylation of formononetin to calycosin. Furthermore, this study successfully achieved the heterologous synthesis of calycosin precursors in yeast and tobacco, confirming the functional expression of these enzymes in plant and microbial hosts. Tissue-specific expression analysis showed differential expression of AmIFS and AmI3′H in roots, stems, and leaves of A. membranaceus , which was positively correlated with formononetin accumulation. These findings not only completed the calycosin biosynthetic pathway, but also laid a theoretical foundation and provided technical support for the scalable production of bioactive isoflavones via synthetic biology.",
  "abstract_shown": true,
  "affiliations": [
    "State Key Laboratory of Discovery and Utilization of Functional Components in Traditional Chinese Medicine, Key Laboratory of Chemical Biology (Ministry of Education), School of Pharmaceutical Sciences, Cheeloo College of Medicine, Shandong University, Jinan, 250012, China",
    "Corresponding author."
  ],
  "article_type": null,
  "arxiv": null,
  "author_total": 12,
  "authors": [
    {
      "affs": [
        0
      ],
      "name": "Ming-Xin Cui"
    },
    {
      "affs": [
        0
      ],
      "name": "Yue Hao"
    },
    {
      "affs": [
        0
      ],
      "name": "Jiang-Nan Li"
    },
    {
      "affs": [
        0
      ],
      "name": "Dan-Dan Xu"
    },
    {
      "affs": [
        0
      ],
      "name": "Pei-Xi Hao"
    },
    {
      "affs": [
        0
      ],
      "name": "Jie Fu"
    },
    {
      "affs": [
        0
      ],
      "name": "Ying Lu"
    },
    {
      "affs": [
        0
      ],
      "name": "Ya-Tong Li"
    },
    {
      "affs": [
        0
      ],
      "name": "Lin-Lin Zhi"
    },
    {
      "affs": [
        0
      ],
      "name": "Jiao-Zhen Zhang"
    },
    {
      "affs": [
        0
      ],
      "name": "Hong-Xiang Lou"
    },
    {
      "affs": [
        1
      ],
      "name": "Ai-Xia Cheng"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/7e022mwsw9mej.bib",
  "cited_by": 0,
  "doi": "10.1016/j.pscia.2026.100118",
  "fulltext_url": "http://www.sciencedirect.com/science/article/pii/S2773216926000115",
  "id": "7e022mwsw9mej",
  "issns": [
    "2773-2169"
  ],
  "landing_url": "https://doi.org/10.1016/j.pscia.2026.100118",
  "language": "en",
  "licence": "CC-BY",
  "licence_label": "CC BY",
  "licence_source": "journal",
  "licence_url": "https://creativecommons.org/licenses/by/4.0/",
  "metadata_licence": null,
  "native_subjects": [
    [
      "lcc",
      "RM1-950"
    ],
    [
      "lcc",
      "RS1-441"
    ],
    [
      "openalex",
      "1312|Plant Gene Expression Analysis"
    ]
  ],
  "openalex": "W7141073551",
  "pdf_url": null,
  "pmcid": null,
  "pmid": null,
  "published": "2026-11-06",
  "publisher": "Elsevier",
  "ris": "https://research.draconitic.com/paper/7e022mwsw9mej.ris",
  "sources": [
    {
      "first_seen": "2026-11-06T14:21:05Z",
      "native_id": "068a4300e76f406abc17d2c13945a740",
      "source": "doaj",
      "url": "https://doi.org/10.1016/j.pscia.2026.100118"
    }
  ],
  "subjects": [
    {
      "field": "biochemistry",
      "origin": "openalex:1312|Plant Gene Expression Analysis",
      "subfield": "molecular-biology"
    }
  ],
  "title": "Functional identification of key enzymes in calycosin biosynthesis in",
  "title_text": "Functional identification of key enzymes in calycosin biosynthesis in",
  "type": "peer-reviewed",
  "updated_at": "2026-11-08T14:20:18Z",
  "url": "https://research.draconitic.com/paper/7e022mwsw9mej",
  "venue": "Pharmaceutical Science Advances",
  "versions": []
}