Energizing eukaryotic cell-free protein synthesis with glucose metabolism
| Autor: | Michael C. Jewett, Jessica C. Stark, Mark J. Anderson, C. Eric Hodgman |
|---|---|
| Jazyk: | angličtina |
| Předmět: |
Saccharomyces cerevisiae
Biophysics Creatine Biochemistry Article chemistry.chemical_compound Adapter molecule crk Adenosine Triphosphate In vitro transcription and translation Structural Biology Natural energy metabolism Genetics Protein biosynthesis Luciferase Molecular Biology Cell-free protein synthesis biology Cell-Free System Cell Biology Phosphate biology.organism_classification Cell biology Glucose chemistry Cell-free biology Protein Biosynthesis biology.protein Protein expression Creatine kinase |
| Zdroj: | FEBS Letters. (15):1723-1727 |
| ISSN: | 0014-5793 |
| DOI: | 10.1016/j.febslet.2015.05.045 |
| Popis: | 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±0.35μgmL−1 active luciferase in batch reactions with 16mM glucose and 25mM 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. |
| Databáze: | OpenAIRE |
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