Maximizing Heterologous Expression of Engineered Type I Polyketide Synthases: Investigating Codon Optimization Strategies
ACS Synthetic Biology, vol. 12, pp. 3366–3380
Abstract
High Resolution Image Download MS PowerPoint Slide Type I polyketide synthases (T1PKSs) hold enormous potential as a rational production platform for the biosynthesis of specialty chemicals. However, despite great progress in this field, the heterologous expression of PKSs remains a major challenge. One of the first measures to improve heterologous gene expression can be codon optimization. Although controversial, choosing the wrong codon optimization strategy can have detrimental effects on the protein and product levels. In this study, we analyzed 11 different codon variants of an engineered T1PKS and investigated in a systematic approach their influence on heterologous expression in Corynebacterium glutamicum, Escherichia coli, and Pseudomonas putida . Our best performing codon variants exhibited a minimum 50-fold increase in PKS protein levels, which also enabled the production of an unnatural polyketide in each of these hosts. Furthermore, we developed a free online tool ( https://basebuddy.lbl.gov ) that offers transparent and highly customizable codon optimization with up-to-date codon usage tables. In this work, we not only highlight the significance of codon optimization but also establish the groundwork for the high-throughput assembly and characterization of PKS pathways in alternative hosts.
Authors 12
-
QB3 · Lawrence Berkeley National Laboratory · Joint BioEnergy Institute · RWTH Aachen University · University of California, Berkeley
Affiliation as printed
Biological Systems & Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, California 94720, United States
California Institute for Quantitative Biosciences (QB3), University of California, Berkeley, California 94720, United States
Institute of Applied Microbiology (iAMB), Aachen Biology and Biotechnology (ABBt), RWTH Aachen University, 52062 Aachen, Germany
Joint BioEnergy Institute, 5885 Hollis Street, Emeryville, California 94608, United States
-
QB3 · Lawrence Berkeley National Laboratory · Joint BioEnergy Institute · University of California, Berkeley
Affiliation as printed
Biological Systems & Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, California 94720, United States
California Institute for Quantitative Biosciences (QB3), University of California, Berkeley, California 94720, United States
Joint BioEnergy Institute, 5885 Hollis Street, Emeryville, California 94608, United States
-
Lawrence Berkeley National Laboratory · Joint BioEnergy Institute
Affiliation as printed
Biological Systems & Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, California 94720, United States
Joint BioEnergy Institute, 5885 Hollis Street, Emeryville, California 94608, United States
-
Lawrence Berkeley National Laboratory · Joint BioEnergy Institute · University of California, Berkeley
Affiliation as printed
Biological Systems & Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, California 94720, United States
Joint BioEnergy Institute, 5885 Hollis Street, Emeryville, California 94608, United States
Joint Program in Bioengineering, University of California, Berkeley, California 94720, United States
-
Lawrence Berkeley National Laboratory · Joint BioEnergy Institute · University of California, Berkeley
Affiliation as printed
Biological Systems & Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, California 94720, United States
Department of Chemical and Biomolecular Engineering, University of California, Berkeley, California 94720, United States
Joint BioEnergy Institute, 5885 Hollis Street, Emeryville, California 94608, United States
-
Lawrence Berkeley National Laboratory · Joint BioEnergy Institute · University of California, Berkeley
Affiliation as printed
Biological Systems & Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, California 94720, United States
Department of Chemical and Biomolecular Engineering, University of California, Berkeley, California 94720, United States
Joint BioEnergy Institute, 5885 Hollis Street, Emeryville, California 94608, United States
-
Lawrence Berkeley National Laboratory · Joint BioEnergy Institute · University of California, Berkeley
Affiliation as printed
Biological Systems & Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, California 94720, United States
Department of Chemical and Biomolecular Engineering, University of California, Berkeley, California 94720, United States
Joint BioEnergy Institute, 5885 Hollis Street, Emeryville, California 94608, United States
-
Lawrence Berkeley National Laboratory · Joint BioEnergy Institute
Affiliation as printed
Biological Systems & Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, California 94720, United States
Joint BioEnergy Institute, 5885 Hollis Street, Emeryville, California 94608, United States
-
Lawrence Berkeley National Laboratory · Joint BioEnergy Institute
Affiliation as printed
Biological Systems & Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, California 94720, United States
Joint BioEnergy Institute, 5885 Hollis Street, Emeryville, California 94608, United States
-
Lawrence Berkeley National Laboratory · Joint BioEnergy Institute
Affiliation as printed
Biological Systems & Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, California 94720, United States
Joint BioEnergy Institute, 5885 Hollis Street, Emeryville, California 94608, United States
-
Affiliation as printed
Institute of Applied Microbiology (iAMB), Aachen Biology and Biotechnology (ABBt), RWTH Aachen University, 52062 Aachen, Germany
-
Jay D. Keasling corresponding
Lawrence Berkeley National Laboratory · Novo Nordisk Foundation · Shenzhen Institutes of Advanced Technology · Joint BioEnergy Institute · University of California, Berkeley · Technical University of Denmark
Affiliation as printed
Biological Systems & Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, California 94720, United States
Department of Chemistry, University of California, Berkeley, California 94720, United States
Environmental Genomics and Systems Biology Division, Lawrence Berkeley National Laboratory, Berkeley, California 94720, United States
Joint BioEnergy Institute, 5885 Hollis Street, Emeryville, California 94608, United States
Joint Program in Bioengineering, University of California, Berkeley, California 94720, United States
Center for Synthetic Biochemistry, Institute for Synthetic Biology, Shenzhen Institutes for Advanced Technologies, Shenzhen 518071, China
Department of Chemical and Biomolecular Engineering, University of California, Berkeley, California 94720, United States
The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, 2800 Kgs. Lyngby, Denmark
Cited by 43 stored of 43
No patents citing this paper on Lens.org (checked 2026-10-06).