{
  "abstract": "Photoenzymes enable stereoselective excited-state transformations beyond the chemical space accessible to natural enzymes, yet their development remains largely dependent on repurposing few privileged protein scaffolds and extensive directed evolution. Although de novo protein design enables tailor-made catalytic architectures, its prevailing transition-state stabilization paradigm is not readily transferable to photoenzymes. Here we establish a substrate-recognition-guided strategy for de novo photoenzyme design, in which the computational construction of substrate-tailored catalytic environments determines stereoselectivity and the genetic incorporation of abiotic thioxanthone photosensitizer endows excited-state reactivity. This approach generated customized photoenzymes for the deracemization of 4-alkenylcyclohexanols that exhibit nanomolar substrate affinity, catalytic efficiencies exceeding 1.5 × 105 M-1 s-1, and up to 99% enantiomeric excess. Crystal structure and kinetic analyses validate the designed model, revealing substrate-recognition-dependent enantioselective energy transfer within the tailored active site. This work establishes substrate recognition as a general design principle for extending de novo enzyme design to excited-state biocatalysis.",
  "abstract_shown": true,
  "affiliations": [
    "State Key Laboratory of Materials Processing and Die and Mould Technology, Key Laboratory of Material Chemistry for Energy Conversion and Storage, Ministry of Education, National Engineering Research Center for Nanomedicine, Hubei Key Laboratory of Bioinorganic Chemistry & Materia Medica, School of Chemistry and Chemical Engineering",
    "Huazhong University of Science and Technology (HUST",
    "Department of Chemistry, Zhejiang Key Laboratory of Low-Carbon Synthesis of Value-Added Chemicals",
    "Zhejiang University",
    "Key Laboratory of Synthetic and Natural Functional Molecule of the Ministry of Education, College of Chemistry and Materials Science",
    "Northwest University",
    "Union Hospital",
    "Tongji Medical College"
  ],
  "article_type": null,
  "arxiv": null,
  "author_total": 17,
  "authors": [
    {
      "affs": [
        0,
        1
      ],
      "name": "Zehui Wang"
    },
    {
      "affs": [
        0,
        1
      ],
      "name": "Yangjinxiu Zhou"
    },
    {
      "affs": [
        2,
        3
      ],
      "name": "Yunhao Li"
    },
    {
      "affs": [
        0,
        1
      ],
      "name": "Guojiao Wu"
    },
    {
      "affs": [
        4,
        5
      ],
      "name": "Mengwen Shi"
    },
    {
      "affs": [
        4,
        5
      ],
      "name": "Siyuan Wang"
    },
    {
      "affs": [
        0,
        1
      ],
      "name": "Tengfei Pang"
    },
    {
      "affs": [
        0,
        1
      ],
      "name": "Yuqi Peng"
    },
    {
      "affs": [
        0,
        1
      ],
      "name": "Jundi Guo"
    },
    {
      "affs": [
        0,
        1
      ],
      "name": "Huiyi Zhou"
    },
    {
      "affs": [
        0,
        1
      ],
      "name": "Yingru Xu"
    },
    {
      "affs": [
        0,
        1
      ],
      "name": "Hongmei Liu"
    },
    {
      "affs": [
        2,
        3
      ],
      "name": "Mowei Zhou"
    },
    {
      "affs": [
        4,
        5
      ],
      "name": "Xi Chen"
    },
    {
      "affs": [
        2,
        3
      ],
      "name": "Pengfei Ji"
    },
    {
      "affs": [
        0,
        1
      ],
      "name": "Fangrui Zhong"
    },
    {
      "affs": [
        0,
        1,
        6,
        7
      ],
      "name": "Yuzhou Wu",
      "orcid": "0000-0003-3229-4982"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/4kbsjygmh0rfy.bib",
  "cited_by": 0,
  "doi": "10.26434/chemrxiv.15010510/v1",
  "fulltext_url": null,
  "id": "4kbsjygmh0rfy",
  "issns": [],
  "landing_url": "https://chemrxiv.org/doi/full/10.26434/chemrxiv.15010510/v1",
  "language": null,
  "licence": "CC-BY",
  "licence_label": "CC BY",
  "licence_source": "journal",
  "licence_url": "https://creativecommons.org/licenses/by/4.0/",
  "metadata_licence": null,
  "native_subjects": [
    [
      "openalex",
      "1312|Enzyme Catalysis and Immobilization"
    ]
  ],
  "openalex": "W7222282076",
  "pdf_url": "https://chemrxiv.org/doi/pdf/10.26434/chemrxiv.15010510/v1",
  "pmcid": null,
  "pmid": null,
  "published": "2026-10-15",
  "publisher": "American Chemical Society (ACS)",
  "ris": "https://research.draconitic.com/paper/4kbsjygmh0rfy.ris",
  "sources": [
    {
      "first_seen": "2026-10-30T12:17:24Z",
      "native_id": "10.26434/chemrxiv.15010510",
      "source": "chemrxiv",
      "url": "https://chemrxiv.org/doi/full/10.26434/chemrxiv.15010510/v1"
    }
  ],
  "subjects": [
    {
      "field": "biochemistry",
      "origin": "openalex:1312|Enzyme Catalysis and Immobilization",
      "subfield": "molecular-biology"
    }
  ],
  "title": "De novo design of photoenzymes with substrate-recognition-encoded enantioselectivity",
  "title_text": "De novo design of photoenzymes with substrate-recognition-encoded enantioselectivity",
  "type": "preprint",
  "updated_at": "2026-10-30T12:17:24Z",
  "url": "https://research.draconitic.com/paper/4kbsjygmh0rfy",
  "venue": "ChemRxiv",
  "versions": []
}