{
  "abstract": "Poly(ester-urethane) bioconversion requires efficient polymer-degrading enzymes and microbial hosts capable of processing the resulting monomers. However, the optimal modular design of secreted cutinases for depolymerizing insoluble polyester-polyurethanes remains largely unexplored. Here, we established a combinatorial synthetic biology strategy in Yarrowia lipolytica based on the fungal cutinase FoCut5a from Fusarium oxysporum. A library of 96 Golden Gate-assembled constructs, combining four secretion signals, four interdomain linkers, and six substrate-binding domains, was integrated site-specifically into the genome and screened for depolymerization of the polyester-polyurethane dispersion Impranil DLN-SD. The best-performing variant fused FoCut5a to the carbohydrate-binding module TrCBM1. LC-MS analysis demonstrated release of adipic acid, 1,6-hexanediol, and neopentyl glycol, confirming ester-bond hydrolysis within the polyester domains of the polymer and additionally detected 6-hydroxycaproate, indicating oxidation of polyurethane-derived 1,6-hexanediol by Y. lipolytica. Purified FoCut5a-TrCBM1 displayed unchanged activity on p-nitrophenyl butyrate but reduced Impranil turbidity 5.6-fold faster and increased release of 1,6-hexanediol and adipic acid by 76-fold and 220-fold, respectively, compared to the unfused enzyme. These results highlight modular enzyme engineering as an effective strategy to enhance polyester-polyurethane deconstruction and support Y. lipolytica as a platform for polymer bioconversion and upcycling.",
  "abstract_shown": true,
  "affiliations": [
    "Department of Biosciences, Biotechnologies and Environment, University of Bari Aldo Moro, Via E. Orabona 4, 70125 Bari, Italy"
  ],
  "article_type": "New results",
  "arxiv": null,
  "author_total": 9,
  "authors": [
    {
      "name": "E. Messina"
    },
    {
      "name": "M. Papi"
    },
    {
      "name": "S. Barile"
    },
    {
      "name": "R. De Rosa"
    },
    {
      "name": "P. Scarcia"
    },
    {
      "name": "A. Biundo"
    },
    {
      "name": "I. Pisano"
    },
    {
      "name": "E. Topakas"
    },
    {
      "affs": [
        0
      ],
      "name": "G. Agrimi"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/epebh4dmdqd1d.bib",
  "cited_by": 0,
  "doi": "10.64898/2026.10.15.759203",
  "fulltext_url": "https://www.biorxiv.org/content/10.64898/2026.10.15.759203v1.full",
  "id": "epebh4dmdqd1d",
  "issns": [],
  "landing_url": "https://www.biorxiv.org/content/10.64898/2026.10.15.759203v1",
  "language": null,
  "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": [
    [
      "biorxiv",
      "synthetic biology"
    ],
    [
      "openalex",
      "1312|Enzyme Catalysis and Immobilization"
    ]
  ],
  "openalex": "W7222473233",
  "pdf_url": "https://www.biorxiv.org/content/10.64898/2026.10.15.759203v1.full.pdf",
  "pmcid": null,
  "pmid": null,
  "published": "2026-10-15",
  "publisher": null,
  "ris": "https://research.draconitic.com/paper/epebh4dmdqd1d.ris",
  "sources": [
    {
      "first_seen": "2026-10-30T12:28:05Z",
      "native_id": "10.64898/2026.10.15.759203",
      "source": "biorxiv",
      "url": "https://www.biorxiv.org/content/10.64898/2026.10.15.759203v1"
    }
  ],
  "subjects": [
    {
      "field": "biology",
      "origin": "biorxiv:synthetic biology",
      "subfield": "systems-synthetic"
    }
  ],
  "title": "Synthetic biology-guided modular engineering of the secreted cutinase FoCut5a with a carbohydrate-binding module improves poly(ester-urethane) depolymerization in Yarrowia lipolytica",
  "title_text": "Synthetic biology-guided modular engineering of the secreted cutinase FoCut5a with a carbohydrate-binding module improves poly(ester-urethane) depolymerization in Yarrowia lipolytica",
  "type": "preprint",
  "updated_at": "2026-10-30T12:28:05Z",
  "url": "https://research.draconitic.com/paper/epebh4dmdqd1d",
  "venue": "bioRxiv",
  "versions": []
}