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Energizing eukaryotic cell-free protein synthesis with glucose metabolism.

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 ?g mL(-1) active luciferase in batch reactions with 16 mM glucose and 25 mM phosphate, resulting in a 16% increase in relative protein yield (?g protein/$ reagents) compared to the CrP/CrK system. Our demonstration provides the foundation for development of cost-effective eukaryotic CFPS platforms.

SUBMITTER: Anderson MJ

PROVIDER: S-EPMC4651010 | biostudies-literature | 2015 Jul

REPOSITORIES: biostudies-literature

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Energizing eukaryotic cell-free protein synthesis with glucose metabolism.

Anderson Mark J MJ Stark Jessica C JC Hodgman C Eric CE Jewett Michael C MC

FEBS letters 20150606 15

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, res ...[more]

PMID: 26054976

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