{
  "abstract": "Enzyme cascades enable complex biochemical transformations, but optimizing them requires searching high-dimensional spaces of enzyme stoichiometry, reaction conditions, and buffer composition. Here we introduce CascadeMAP, an autonomous microfluidic platform for closed-loop optimization of enzyme cascades, integrating high-throughput microfluidics with Bayesian optimization and assisted by agentic analyses. We demonstrate the platform across two three-enzyme cascades: (i) a glycerol detection pathway monitored by fluorescence and (ii) a 1,2,3-trichloropropane degradation pathway monitored by label-free Raman spectroscopy, providing orthogonal detection modalities. Bayesian optimization reached comparable optima with threefold fewer experimental conditions than Design of Experiments. A multi-agent AI system automated processing 2.1 GB of experimental data, pattern recognition, and insight synthesis. Operating without human intervention for 7 days, CascadeMAP processed ~295,000 reactions across ~7,400 different conditions. This capability establishes a generalizable, resource-efficient framework for autonomously optimizing enzyme cascades and metabolic pathways and accelerates the development of biocatalytic and synthetic biological systems.",
  "abstract_shown": true,
  "affiliations": [
    "Masaryk University"
  ],
  "article_type": "New results",
  "arxiv": null,
  "author_total": 23,
  "authors": [
    {
      "name": "M. Vasina"
    },
    {
      "name": "D. Kovar"
    },
    {
      "name": "M. Kizovsky"
    },
    {
      "name": "D. Lacko"
    },
    {
      "name": "P. Vanacek"
    },
    {
      "name": "M. Herich"
    },
    {
      "name": "E. Volf"
    },
    {
      "name": "L. Drdla"
    },
    {
      "name": "S. Cabalova"
    },
    {
      "name": "P. Sikorova"
    },
    {
      "name": "M. Jirasek"
    },
    {
      "name": "P. Solansky"
    },
    {
      "name": "J. Jezek"
    },
    {
      "name": "O. Samek"
    },
    {
      "name": "F. Dousek"
    },
    {
      "name": "H. Walner"
    },
    {
      "name": "P. Zemanek"
    },
    {
      "name": "A. deMello"
    },
    {
      "name": "Z. Pilat"
    },
    {
      "name": "J. Damborsky"
    },
    {
      "name": "S. Stavrakis"
    },
    {
      "name": "S. Mazurenko"
    },
    {
      "affs": [
        0
      ],
      "name": "Z. Prokop"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/8zezm09v7sre9.bib",
  "cited_by": 2,
  "doi": "10.64898/2026.06.04.730034",
  "fulltext_url": "https://www.biorxiv.org/content/10.64898/2026.06.04.730034v2.full",
  "id": "8zezm09v7sre9",
  "issns": [],
  "landing_url": "https://www.biorxiv.org/content/10.64898/2026.06.04.730034v2",
  "language": null,
  "licence": "CC-BY",
  "licence_label": "CC BY",
  "licence_source": "article",
  "licence_url": "https://creativecommons.org/licenses/by/4.0/",
  "metadata_licence": null,
  "native_subjects": [
    [
      "biorxiv",
      "biochemistry"
    ],
    [
      "openalex",
      "1312|Enzyme Catalysis and Immobilization"
    ]
  ],
  "openalex": "W7163806566",
  "pdf_url": "https://www.biorxiv.org/content/10.64898/2026.06.04.730034v2.full.pdf",
  "pmcid": null,
  "pmid": null,
  "published": "2026-06-04",
  "publisher": null,
  "ris": "https://research.draconitic.com/paper/8zezm09v7sre9.ris",
  "sources": [
    {
      "first_seen": "2026-10-30T12:29:00Z",
      "native_id": "10.64898/2026.06.04.730034",
      "source": "biorxiv",
      "url": "https://www.biorxiv.org/content/10.64898/2026.06.04.730034v2"
    }
  ],
  "subjects": [
    {
      "field": "biochemistry",
      "origin": "openalex:1312|Enzyme Catalysis and Immobilization",
      "subfield": "molecular-biology"
    }
  ],
  "title": "CascadeMAP: Autonomous Closed-loop Optimization of Enzyme Cascades via Microfluidics, Machine Learning and Agentic AI",
  "title_text": "CascadeMAP: Autonomous Closed-loop Optimization of Enzyme Cascades via Microfluidics, Machine Learning and Agentic AI",
  "type": "preprint",
  "updated_at": "2026-10-30T12:29:00Z",
  "url": "https://research.draconitic.com/paper/8zezm09v7sre9",
  "venue": "bioRxiv",
  "versions": []
}