{
  "abstract": "Umbelliferone, a key precursor for the biosynthesis of high-value furanocoumarin derivatives, is produced via the shikimate pathway. The oleaginous yeast Yarrowia lipolytica possesses an inherently robust shikimate pathway; however, de novo biosynthesis of umbelliferone in this microbial chassis has not been realized to date. Low intracellular pools of p-coumaric acid and suboptimal catalytic activity of downstream hydroxylation and cyclization enzymes represent the major bottlenecks limiting efficient umbelliferone production. In this study, we systematically screened five PAL/TAL variants, three 4CL isoforms, and two C2′H enzymes derived from diverse plant and microbial sources. The bifunctional enzyme RtPAL-TAL exhibited the highest productivity of p-coumaric acid, while the combination of At4CL3 and RgC2′H achieved the maximum umbelliferone accumulation. Expression of feedback-insensitive shikimate pathway mutant enzymes effectively relieved intrinsic pathway feedback inhibition, resulting in a 10.2-fold improvement in p-coumaric acid titer. All synthetic biosynthetic gene cassettes were stably integrated into the Y. lipolytica genome to generate the engineered strain YL-PU, which yielded 3.29 mg/L umbelliferone in shake-flask cultures. Further fed-batch fermentation in a 5 L bioreactor increased the umbelliferone titer to 125.52 mg/L at 84 h of cultivation. This work establishes the first de novo umbelliferone biosynthesis pathway in Y. lipolytica and identifies that C2′H represents a major candidate bottleneck under the tested configuration, in agreement with earlier findings. These findings provide a robust foundation for further metabolic engineering of the Y. lipolytica chassis to enable high-titer production of coumarins and other phenylpropanoid natural products.",
  "abstract_shown": true,
  "affiliations": [
    "Shanghai Key Laboratory of Bio-Energy Crops, School of Life Sciences, Shanghai University, Shanghai 200444, China"
  ],
  "article_type": null,
  "arxiv": null,
  "author_total": 5,
  "authors": [
    {
      "affs": [
        0
      ],
      "name": "Fengliu Lu"
    },
    {
      "affs": [
        0
      ],
      "name": "Ying Luo"
    },
    {
      "affs": [
        0
      ],
      "name": "Zhen Lei"
    },
    {
      "affs": [
        0
      ],
      "name": "Changfu Li"
    },
    {
      "affs": [
        0
      ],
      "name": "Yansheng Zhang",
      "orcid": "0000-0001-7597-9821"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/ds4t5dvx043xr.bib",
  "cited_by": 0,
  "doi": "10.3390/biom16101450",
  "fulltext_url": null,
  "id": "ds4t5dvx043xr",
  "issns": [
    "2218-273X"
  ],
  "landing_url": "https://www.mdpi.com/2218-273X/16/10/1450",
  "language": null,
  "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|Microbial Metabolic Engineering and Bioproduction"
    ]
  ],
  "openalex": "W7220356424",
  "pdf_url": null,
  "pmcid": null,
  "pmid": null,
  "published": "2026-10-04",
  "publisher": "MDPI AG",
  "ris": "https://research.draconitic.com/paper/ds4t5dvx043xr.ris",
  "sources": [
    {
      "first_seen": "2026-10-30T12:18:59Z",
      "native_id": "10.3390/biom16101450",
      "source": "crossref",
      "url": "https://www.mdpi.com/2218-273X/16/10/1450"
    }
  ],
  "subjects": [
    {
      "field": "biochemistry",
      "origin": "openalex:1312|Microbial Metabolic Engineering and Bioproduction",
      "subfield": "molecular-biology"
    }
  ],
  "title": "Metabolic Engineering of Yarrowia lipolytica for De Novo Biosynthesis of Umbelliferone",
  "title_text": "Metabolic Engineering of Yarrowia lipolytica for De Novo Biosynthesis of Umbelliferone",
  "type": "peer-reviewed",
  "updated_at": "2026-10-30T12:18:59Z",
  "url": "https://research.draconitic.com/paper/ds4t5dvx043xr",
  "venue": "Biomolecules",
  "versions": []
}