{
  "abstract": "To address the persistent challenges of low yield and significant batch-to-batch variability inherent in the traditional plant-based extraction of horseradish peroxidase (HRP), this study aimed to develop an efficient production strategy for HRP-C through rational molecular design and process optimization. A recombinant engineering strain was constructed in Pichia pastoris GS115, and site-directed mutagenesis was performed based on FoldX virtual screening to evaluate mutations affecting enzyme stability and catalytic properties. A comprehensive 50 L scale process was established, integrating high-density fermentation, purification, and lyophilized formulation. Results indicated that exogenous supplementation with 5 μM hemin significantly enhanced enzymatic activity. The HRP-C-D282I variant exhibited a 17.7% increase in specific activity and a 2.14-fold extension of half-life at 60 °C compared with the wild-type enzyme. Furthermore, Endo H-mediated deglycosylation was associated with a dose-dependent decrease in HRP-C-D282I activity, suggesting that N-glycosylation contributes to enzyme performance under the conditions tested. The optimized process achieved a recombinant protein concentration of 216 mg/L during fermentation and a purified enzyme specific activity of 2600 U/mg, with a lyophilized powder reconstitution time of less than 60 s. This study provides a proof-of-concept basis for the potential industrial production and application of recombinant HRP.",
  "abstract_shown": true,
  "affiliations": [
    "National Biopesticide Engineering Technology Research Center, Hubei Biopesticide Engineering Research Center, Hubei Academy of Agricultural Sciences, Biopesticide Branch of Hubei Innovation Centre of Agricultural Science and Technology, Wuhan 430064, China",
    "State Key Laboratory of Biocatalysis and Enzyme, Engineering Hubei Collaborative Innovation Center for Green Transformation of Bio-Resources, Hubei Key Laboratory of Industrial Biotechnology, College of Life Sciences of Hubei University, Hubei University, Wuhan 430062, China"
  ],
  "article_type": "research article",
  "arxiv": null,
  "author_total": 6,
  "authors": [
    {
      "affs": [
        0,
        1
      ],
      "name": "Juran Cui"
    },
    {
      "affs": [
        0,
        1
      ],
      "name": "Qifeng Yu"
    },
    {
      "affs": [
        1
      ],
      "name": "Sen Yang"
    },
    {
      "affs": [
        0,
        1
      ],
      "name": "Quanxi Wang"
    },
    {
      "affs": [
        1
      ],
      "name": "Yaping Wang"
    },
    {
      "affs": [
        0
      ],
      "name": "Ben Rao"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/bqcsh4bg9jay0.bib",
  "cited_by": 0,
  "doi": "10.3390/molecules31203730",
  "fulltext_url": "https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13707735/?tool=EBI",
  "id": "bqcsh4bg9jay0",
  "issns": [
    "1420-3049"
  ],
  "landing_url": "https://www.mdpi.com/1420-3049/31/20/3730",
  "language": "eng",
  "licence": "CC-BY-4.0",
  "licence_label": "CC BY 4.0",
  "licence_source": "article",
  "licence_url": "https://creativecommons.org/licenses/by/4.0/",
  "metadata_licence": null,
  "native_subjects": [
    [
      "openalex",
      "1312|Enzyme Catalysis and Immobilization"
    ]
  ],
  "openalex": "W7222956129",
  "pdf_url": "https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13707735/pdf/?tool=EBI",
  "pmcid": "PMC13707735",
  "pmid": null,
  "published": "2026-10-01",
  "publisher": "MDPI AG",
  "ris": "https://research.draconitic.com/paper/bqcsh4bg9jay0.ris",
  "sources": [
    {
      "first_seen": "2026-10-30T12:18:42Z",
      "native_id": "10.3390/molecules31203730",
      "source": "crossref",
      "url": "https://www.mdpi.com/1420-3049/31/20/3730"
    },
    {
      "first_seen": "2026-10-30T12:21:46Z",
      "native_id": "PMC:PMC13707735",
      "source": "europepmc",
      "url": "https://europepmc.org/article/PMC/PMC13707735"
    }
  ],
  "subjects": [
    {
      "field": "biochemistry",
      "origin": "openalex:1312|Enzyme Catalysis and Immobilization",
      "subfield": "molecular-biology"
    }
  ],
  "title": "Rational Engineering and Preparation of Horseradish Peroxidase C",
  "title_text": "Rational Engineering and Preparation of Horseradish Peroxidase C",
  "type": "peer-reviewed",
  "updated_at": "2026-10-30T12:21:46Z",
  "url": "https://research.draconitic.com/paper/bqcsh4bg9jay0",
  "venue": "Molecules",
  "versions": []
}