Biological and Medicinal Chemistry

A Single Polyphosphate Kinase-Based NTP Regeneration System Driving Cell-Free Protein Synthesis

Po-Hsiang Wang Earth-Life Science Institute (ELSI), Tokyo Institute of Technology


Current reconstituted cell-free protein synthesis systems regenerate the ATP and GTP required for peptide bond formation using phosphocreatine and three coupled kinase reactions. Here, we replace the three-kinase system with a single polyphosphate kinase, resulting in a comparable protein yield and enabling functional expression of a heat-sensitive enzyme at 37°C.


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Supplementary material

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Supporting Information V2