Maximizing Heterologous Expression of Engineered Type I Polyketide Synthases: Investigating Codon Optimization Strategies
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- Joint BioEnergy Institute, 5885 Hollis Street, Emeryville, California 94608, United States, Biological Systems & Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, California 94720, United States, 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, California 94720, United States
- Joint BioEnergy Institute, 5885 Hollis Street, Emeryville, California 94608, United States, 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, Biological Systems & Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, California 94720, United States
- Joint BioEnergy Institute, 5885 Hollis Street, Emeryville, California 94608, United States, Biological Systems & Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, California 94720, United States, Joint Program in Bioengineering, University of California, Berkeley, California 94720, United States
- Joint BioEnergy Institute, 5885 Hollis Street, Emeryville, California 94608, United States, 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
- 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, Biological Systems & Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, California 94720, United States, Joint Program in Bioengineering, University of California, 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
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.
- Research Organization:
- Univ. of California, Berkeley, CA (United States)
- Sponsoring Organization:
- USDOE Office of Science (SC), Biological and Environmental Research (BER); USDOE Office of Energy Efficiency and Renewable Energy (EERE), Office of Sustainable Transportation. Bioenergy Technologies Office (BETO)
- Grant/Contract Number:
- AC02-05CH11231; EE0008926
- OSTI ID:
- 2202869
- Alternate ID(s):
- OSTI ID: 2217103; OSTI ID: 2217547
- Journal Information:
- ACS Synthetic Biology, Journal Name: ACS Synthetic Biology Vol. 12 Journal Issue: 11; ISSN 2161-5063
- Publisher:
- American Chemical SocietyCopyright Statement
- Country of Publication:
- United States
- Language:
- English
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