An integrated cell-free metabolic platform for protein production and synthetic biology

Michael C. Jewett, Kara A. Calhoun, Alexei Voloshin, Jessica J. Wuu, James R. Swartz

Research output: Contribution to journalArticlepeer-review

284 Scopus citations


Cell-free systems offer a unique platform for expanding the capabilities of natural biological systems for useful purposes, i.e. synthetic biology. They reduce complexity, remove structural barriers, and do not require the maintenance of cell viability. Cell-free systems, however, have been limited by their inability to co-activate multiple biochemical networks in a single integrated platform. Here, we report the assessment of biochemical reactions in an Escherichia coli cell-free platform designed to activate natural metabolism, the Cytomim system. We reveal that central catabolism, oxidative phosphorylation, and protein synthesis can be co-activated in a single reaction system. Never before have these complex systems been shown to be simultaneously activated without living cells. The Cytomim system therefore promises to provide the metabolic foundation for diverse ab initio cell-free synthetic biology projects. In addition, we describe an improved Cytomim system with enhanced protein synthesis yields (up to 1200 mg/l in 2 h) and lower costs to facilitate production of protein therapeutics and biochemicals that are difficult to make in vivo because of their toxicity, complexity, or unusual cofactor requirements.

Original languageEnglish (US)
Article number220
JournalMolecular Systems Biology
StatePublished - 2008


  • Cell-free biology
  • In vitro translation
  • Oxidative phosphorylation
  • Protein synthesis
  • Synthetic biology

ASJC Scopus subject areas

  • Information Systems
  • General Immunology and Microbiology
  • Applied Mathematics
  • General Biochemistry, Genetics and Molecular Biology
  • General Agricultural and Biological Sciences
  • Computational Theory and Mathematics


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