Abstract
Abstract Eukaryotic cell-free protein synthesis (CFPS) is limited by the dependence on costly high-energy phosphate compounds and exogenous enzymes to power protein synthesis (e.g., creatine phosphate and creatine kinase, CrP/CrK). Here, we report the ability to use glucose as a secondary energy substrate to regenerate ATP in a Saccharomyces cerevisiae crude extract CFPS platform. We observed synthesis of 3.64 ± 0.35 μg mL-1 active luciferase in batch reactions with 16 mM glucose and 25 mM phosphate, resulting in a 16% increase in relative protein yield (μg protein/$ reagents) compared to the CrP/CrK system. Our demonstration provides the foundation for development of cost-effective eukaryotic CFPS platforms.
| Original language | English (US) |
|---|---|
| Article number | 37201 |
| Pages (from-to) | 1723-1727 |
| Number of pages | 5 |
| Journal | FEBS Letters |
| Volume | 589 |
| Issue number | 15 |
| DOIs | |
| State | Published - Jul 3 2015 |
Keywords
- Cell-free biology
- Cell-free protein synthesis
- In vitro transcription and translation
- Natural energy metabolism
- Protein expression
- Saccharomyces cerevisiae
ASJC Scopus subject areas
- Biophysics
- Structural Biology
- Biochemistry
- Molecular Biology
- Genetics
- Cell Biology
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Cite this
Anderson, M. J., Stark, J. C., Hodgman, C. E., & Jewett, M. C. (2015). Energizing eukaryotic cell-free protein synthesis with glucose metabolism. FEBS Letters, 589(15), 1723-1727. [37201]. https://doi.org/10.1016/j.febslet.2015.05.045