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

preprint
revised on 20.08.2019, 07:28 and posted on 20.08.2019, 18:29 by Po-Hsiang Wang, Kosuke Fujishima, Samuel Berhanu, Yutetsu Kuruma, Tony Z. Jia, Anna N. Khusnutdinova, Alexander F. Yakunin, Shawn E. McGlynn

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.

History

Email Address of Submitting Author

tommy.wang@elsi.jp

Institution

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

Country

Japan

ORCID For Submitting Author

0000-0001-9900-0972

Declaration of Conflict of Interest

The authors declare no conflict of interest.

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