Engineering Pseudomonas putida for production of 3-hydroxyacids using hybrid type I polyketide synthases

Matthias Schmidt; Aaron A. Vilchez; Namil Lee; Leah S. Keiser; Allison N. Pearson; Mitchell G. Thompson; Yolanda Zhu; Robert W. Haushalter; Adam M. Deutschbauer; Satoshi Yuzawa; Lars M. Blank; Jay D. Keasling

doi:10.1016/j.mec.2025.e00261

Metabolic Engineering Communications (Jun 2025)

Engineering Pseudomonas putida for production of 3-hydroxyacids using hybrid type I polyketide synthases

Matthias Schmidt,
Aaron A. Vilchez,
Namil Lee,
Leah S. Keiser,
Allison N. Pearson,
Mitchell G. Thompson,
Yolanda Zhu,
Robert W. Haushalter,
Adam M. Deutschbauer,
Satoshi Yuzawa,
Lars M. Blank,
Jay D. Keasling

Affiliations

Matthias Schmidt: Joint BioEnergy Institute, 5885 Hollis Street, Emeryville, CA 94608, USA; Biological Systems & Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, CA, 94720, USA; Institute of Applied Microbiology (iAMB), Aachen Biology and Biotechnology (ABBt), RWTH Aachen University, 52062 Aachen, Germany; California Institute for Quantitative Biosciences (QB3), University of California, Berkeley, CA, 94720, USA
Aaron A. Vilchez: Joint BioEnergy Institute, 5885 Hollis Street, Emeryville, CA 94608, USA; Biological Systems & Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, CA, 94720, USA; Department of Chemical and Biomolecular Engineering, University of California, Berkeley, CA, 94720, USA
Namil Lee: Joint BioEnergy Institute, 5885 Hollis Street, Emeryville, CA 94608, USA; Biological Systems & Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, CA, 94720, USA; California Institute for Quantitative Biosciences (QB3), University of California, Berkeley, CA, 94720, USA
Leah S. Keiser: Joint BioEnergy Institute, 5885 Hollis Street, Emeryville, CA 94608, USA; Biological Systems & Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, CA, 94720, USA; Department of Chemical and Biomolecular Engineering, University of California, Berkeley, CA, 94720, USA
Allison N. Pearson: Joint BioEnergy Institute, 5885 Hollis Street, Emeryville, CA 94608, USA; Biological Systems & Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, CA, 94720, USA; Department of Plant and Microbial Biology, University of California, Berkeley, CA, 94720, USA
Mitchell G. Thompson: Joint BioEnergy Institute, 5885 Hollis Street, Emeryville, CA 94608, USA; Environmental Genomics and Systems Biology Division, Lawrence Berkeley National Laboratory, Berkeley, CA, 94720, USA
Yolanda Zhu: Joint BioEnergy Institute, 5885 Hollis Street, Emeryville, CA 94608, USA; Biological Systems & Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, CA, 94720, USA; Department of Chemical and Biomolecular Engineering, University of California, Berkeley, CA, 94720, USA
Robert W. Haushalter: Joint BioEnergy Institute, 5885 Hollis Street, Emeryville, CA 94608, USA; Biological Systems & Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, CA, 94720, USA
Adam M. Deutschbauer: Department of Plant and Microbial Biology, University of California, Berkeley, CA, 94720, USA; Environmental Genomics and Systems Biology Division, Lawrence Berkeley National Laboratory, Berkeley, CA, 94720, USA
Satoshi Yuzawa: Joint BioEnergy Institute, 5885 Hollis Street, Emeryville, CA 94608, USA; Biological Systems & Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, CA, 94720, USA; Systems Biology Program, Graduate School of Media and Governance, Keio University, Fujisawa, Kanagawa, 252-0882, Japan; Institute for Advanced Biosciences, Keio University, Tsuruoka, Yamagata, 997-0017, Japan
Lars M. Blank: Institute of Applied Microbiology (iAMB), Aachen Biology and Biotechnology (ABBt), RWTH Aachen University, 52062 Aachen, Germany
Jay D. Keasling: Joint BioEnergy Institute, 5885 Hollis Street, Emeryville, CA 94608, USA; Biological Systems & Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, CA, 94720, USA; California Institute for Quantitative Biosciences (QB3), University of California, Berkeley, CA, 94720, USA; Joint Program in Bioengineering, University of California, Berkeley/San Francisco, CA, 94720, USA; Department of Chemistry, University of California, Berkeley, CA, 94720, USA; Environmental Genomics and Systems Biology Division, Lawrence Berkeley National Laboratory, Berkeley, CA, 94720, USA; Department of Chemical and Biomolecular Engineering, University of California, Berkeley, CA, 94720, USA; The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, 2800 Kgs. Lyngby, Denmark; Corresponding author. Joint BioEnergy Institute, 5885 Hollis Street, Emeryville, CA, 94608, USA.

DOI: https://doi.org/10.1016/j.mec.2025.e00261
Journal volume & issue: Vol. 20
p. e00261

Abstract

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Engineered type I polyketide synthases (T1PKSs) are a potentially transformative platform for the biosynthesis of small molecules. Due to their modular nature, T1PKSs can be rationally designed to produce a wide range of bulk or specialty chemicals. While heterologous PKS expression is best studied in microbes of the genus Streptomyces, recent studies have focused on the exploration of non-native PKS hosts. The biotechnological production of chemicals in fast growing and industrial relevant hosts has numerous economic and logistic advantages. With its native ability to utilize alternative feedstocks, Pseudomonas putida has emerged as a promising workhorse for the sustainable production of small molecules. Here, we outline the assessment of P. putida as a host for the expression of engineered T1PKSs and production of 3-hydroxyacids. After establishing the functional expression of an engineered T1PKS, we successfully expanded and increased the pool of available acyl-CoAs needed for the synthesis of polyketides using transposon sequencing and protein degradation tagging. This work demonstrates the potential of T1PKSs in P. putida as a production platform for the sustainable biosynthesis of unnatural polyketides.

Published in Metabolic Engineering Communications

ISSN: 2214-0301 (Online)
Publisher: Elsevier
Country of publisher: Netherlands
LCC subjects: Technology: Chemical technology: Biotechnology; Science: Biology (General)
Website: https://www.sciencedirect.com/journal/metabolic-engineering-communications

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