Engineering controllable alteration of malonyl-CoA levels to enhance polyketide production
Heterologous expression of polyketide synthase (PKS) genes in Escherichia coli has enabled the production of various valuable natural and synthetic products. However, the limited availability of malonyl-CoA (M-CoA) in E. coli remains a substantial impediment to high-titer polyketide production. Here we address this limitation by disrupting the native M-CoA biosynthetic pathway and introducing an orthogonal pathway comprising a malonate transporter and M-CoA ligase, enabling efficient M-CoA biosynthesis under malonate supplementation. This approach substantially increases M-CoA levels, enhancing fatty acid and polyketide titers while reducing the promiscuous activity of PKSs toward undesired acyl-CoA substrates. Subsequent adaptive laboratory evolution of these strains provides insights into M-CoA regulation and identifies mutations that further boost M-CoA and polyketide production. This strategy improves E. coli as a host for polyketide biosynthesis and advances understanding of M-CoA metabolism in microbial systems.
Authors
- Jay D. Keasling (ORCID: https://orcid.org/0000-0003-4170-6088)
- Sarah H. Klass (ORCID: https://orcid.org/0000-0002-1021-5342)
- Jennifer Gin (ORCID: https://orcid.org/0000-0001-5636-7563)
- Christopher J. Petzold (ORCID: https://orcid.org/0000-0002-8270-5228)
- Aidan E. Cowan (ORCID: https://orcid.org/0000-0003-2810-9318)
- Seokjung Cheong (ORCID: https://orcid.org/0000-0002-5699-2278)
- Namil Lee (ORCID: https://orcid.org/0009-0000-0382-8741)
- Nathan Lanclos (ORCID: https://orcid.org/0000-0003-1995-3883)
- Mia Wesselkamper
- Yan Chen (ORCID: https://orcid.org/0009-0009-1387-4165)
- Zilong Wang (ORCID: https://orcid.org/0000-0001-8589-8419)
Institutions
- Berkeley College (US)
- Lawrence Berkeley National Laboratory (US)
- Danish Academy of Technical Sciences (DK)
- Joint BioEnergy Institute (US)
- University of California, Berkeley (US)
- Technical University of Denmark (DK)
Publication Details
- Journal
- Nature Chemical Biology
- Published
- 2025-06-11
- DOI
- https://doi.org/10.1038/s41589-025-01911-6
- Citations
- 8
- Primary Topic
- Fungal and yeast genetics research
- Type
- article
- Field-Weighted Citation Impact
- 6.45
Funders
- National Science Foundation
- U.S. Department of Energy
- Office of Science
- Biological and Environmental Research