{
  "abstract": "The diazo group is a highly valuable functional group in organic synthesis; however, its conventional preparation requires harsh conditions. Here, we provide a systematic analysis of ATP-dependent diazotases from actinomycetes that catalyze the condensation of nitrite with aromatic amines. Sequence similarity network analysis of AMP-binding enzymes revealed two families (type Ia/Ib), with type Ib further classified into 14 groups. In vitro assays confirmed the diazotization activities of the diazotases from 7 groups in the type Ib family, each displaying distinct substrate preferences. Remarkably, diazotases in group 3 exhibited broad substrate specificity and superior catalytic efficiency. The dimeric structure of a representative enzyme, Mco01_40450, was determined at 3.08 Å resolution by cryogenic electron microscopy with C2 symmetry. Subsequent symmetry expansion and 3D classification revealed the structure with AMP, pyrophosphate, and 4-aminohydrocinnamic acid bound to the active site at a resolution of 3.16 Å. Expansion of the substrate-binding pocket and its entrance via amino acid substitution enabled the reaction of bulkier aromatic amines. Unlike classical chemical diazotization, Mco01_40450 operates under mild aqueous conditions and is compatible with sensitive protecting groups. Our findings highlight the structural and functional diversity of diazotases and establish group 3 diazotases as promising, engineerable biocatalysts for selective and efficient diazo installation.",
  "abstract_shown": true,
  "affiliations": [
    "Department of Biotechnology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo113-8657, Japan.",
    "Collaborative Research Institute for Innovative Microbiology, The University of Tokyo, Bunkyo-ku, Tokyo113-8657, Japan."
  ],
  "article_type": "research article",
  "arxiv": null,
  "author_total": 4,
  "authors": [
    {
      "affs": [
        0
      ],
      "name": "Jiayu Ning"
    },
    {
      "affs": [
        0
      ],
      "name": "Seiji Kawai"
    },
    {
      "affs": [
        0,
        1
      ],
      "name": "Yohei Katsuyama",
      "orcid": "0000-0003-4905-5320"
    },
    {
      "affs": [
        0,
        1
      ],
      "name": "Yasuo Ohnishi"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/05eyb8pq4zcee.bib",
  "cited_by": 0,
  "doi": "10.1021/jacs.6c11480",
  "fulltext_url": "https://europepmc.org/articles/PMC13674236",
  "id": "05eyb8pq4zcee",
  "issns": [
    "0002-7863",
    "1520-5126"
  ],
  "landing_url": "https://europepmc.org/article/MED/42913524",
  "language": "eng",
  "licence": "CC-BY-NC-ND",
  "licence_label": "CC BY-NC-ND",
  "licence_source": "article",
  "licence_url": "https://creativecommons.org/licenses/by-nc-nd/4.0/",
  "metadata_licence": null,
  "native_subjects": [
    [
      "openalex",
      "1312|Enzyme Catalysis and Immobilization"
    ]
  ],
  "openalex": "W7202351763",
  "pdf_url": "https://europepmc.org/articles/PMC13674236?pdf=render",
  "pmcid": "PMC13674236",
  "pmid": "42913524",
  "published": "2026-10-01",
  "publisher": null,
  "ris": "https://research.draconitic.com/paper/05eyb8pq4zcee.ris",
  "sources": [
    {
      "first_seen": "2026-10-30T12:20:14Z",
      "native_id": "MED:42913524",
      "source": "europepmc",
      "url": "https://europepmc.org/article/MED/42913524"
    }
  ],
  "subjects": [
    {
      "field": "biochemistry",
      "origin": "openalex:1312|Enzyme Catalysis and Immobilization",
      "subfield": "molecular-biology"
    }
  ],
  "title": "Promiscuous ATP-Dependent Diazotases Discovered by Comprehensive Genome Mining Based on Sequence Similarity Network Analysis.",
  "title_text": "Promiscuous ATP-Dependent Diazotases Discovered by Comprehensive Genome Mining Based on Sequence Similarity Network Analysis.",
  "type": "peer-reviewed",
  "updated_at": "2026-10-30T12:20:14Z",
  "url": "https://research.draconitic.com/paper/05eyb8pq4zcee",
  "venue": "Journal of the American Chemical Society",
  "versions": []
}