{
  "abstract": "There is intense interest in using enzyme biocatalysts for chemically challenging oxidation reactions involving the activation of inert carbon-hydrogen bonds in organic molecules. The cytochrome P450 family of heme monooxygenase enzymes can catalyse selective C-H bond hydroxylations and epoxidation reactions. However, the complex and costly nature of these multicomponent systems and the low stability and activity of these systems make larger scale reactions challenging. Thermophilic P450s from bacteria and archaea have been identified but have low activity when reconstituted using electron transfer partner proteins and surrogate oxygen donors. We demonstrate that two cytochrome P450 enzymes (CYP119A1 and CYP119A2) from hyperthermophilic archaea can be engineered to be efficient peroxygenase enzymes requiring only hydrogen peroxide to function. Catalytic oxidation was achieved through modification of three residues in the dioxygen binding groove, including the important addition of a proline residue after the glutamate acid/base residue which significantly enhanced the activity (8-fold over the double mutant and three orders of magnitude over the natural enzyme). These peroxygenases oxidised tridecanoic acid with high activity and enabled further investigation of the substrate range of these enzymes. Both enzymes could oxidise fatty acids and related compounds as well as styrene and the norisoprenoid β-ionone. The oxidation of these substrates occurred with excellent regioselectivity. The CYP119A2-QEP variant was stable to heating at 85-90 °C, had higher activity at 70 °C, could tolerate up to 20 % v/v acetonitrile and was able to catalyse ≥1000 total turnovers across the range of substrates tested. These new peroxygenase biocatalysts, provide hyperthermostable enzyme scaffolds to enable selective and catalytic oxidation of inert C-H bonds using a single enzyme system and cheap cofactors.",
  "abstract_shown": true,
  "affiliations": [
    "Discipline of Chemistry, School of Physics, Chemistry and Earth Sciences",
    "Adelaide University"
  ],
  "article_type": null,
  "arxiv": null,
  "author_total": 4,
  "authors": [
    {
      "affs": [
        0,
        1
      ],
      "name": "Joel H. Z. Lee",
      "orcid": "0000-0001-6288-9034"
    },
    {
      "affs": [
        0,
        1
      ],
      "name": "Alecia R. Gee"
    },
    {
      "affs": [
        0,
        1
      ],
      "name": "Oghenesivwe Osiebe"
    },
    {
      "affs": [
        0,
        1
      ],
      "name": "Stephen G. Bell"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/2kpep179r9pc6.bib",
  "cited_by": 0,
  "doi": "10.26434/chemrxiv.15008459/v1",
  "fulltext_url": null,
  "id": "2kpep179r9pc6",
  "issns": [],
  "landing_url": "https://chemrxiv.org/doi/full/10.26434/chemrxiv.15008459/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": "W7211966237",
  "pdf_url": "https://chemrxiv.org/doi/pdf/10.26434/chemrxiv.15008459/v1",
  "pmcid": null,
  "pmid": null,
  "published": "2026-09-07",
  "publisher": "American Chemical Society (ACS)",
  "ris": "https://research.draconitic.com/paper/2kpep179r9pc6.ris",
  "sources": [
    {
      "first_seen": "2026-10-30T12:17:17Z",
      "native_id": "10.26434/chemrxiv.15008459",
      "source": "chemrxiv",
      "url": "https://chemrxiv.org/doi/full/10.26434/chemrxiv.15008459/v2"
    }
  ],
  "subjects": [
    {
      "field": "biochemistry",
      "origin": "openalex:1312|Enzyme Catalysis and Immobilization",
      "subfield": "molecular-biology"
    }
  ],
  "title": "Developing Efficient and Hyperthermostable Heme Peroxygenase Enzymes as a Biocatalytic Platform for Selective C-H bond Oxidation Reactions",
  "title_text": "Developing Efficient and Hyperthermostable Heme Peroxygenase Enzymes as a Biocatalytic Platform for Selective C-H bond Oxidation Reactions",
  "type": "preprint",
  "updated_at": "2026-10-30T12:17:24Z",
  "url": "https://research.draconitic.com/paper/2kpep179r9pc6",
  "venue": "ChemRxiv",
  "versions": []
}