Cell-free synthesis is the formation of biological molecules through biochemical reactions carried out by cellular components outside intact living cells. The reaction system contains catalytically active machinery supplied as a cell extract or assembled from purified components, together with substrates, cofactors, salts, and an appropriate source of chemical energy [1]. In systems that synthesize proteins, a nucleic acid template specifies the product, and ribosomes, transfer RNAs, aminoacyl-transfer RNA synthetases, and translation factors mediate polypeptide formation [2]. Enzymes retained in an extract or supplied separately can also form interconnected pathways that convert substrates into small molecules [3]. Reaction activity depends on the availability and regeneration of energy carriers, the biochemical environment, and the accumulation or removal of inhibitory products [4]. The composition and organization of these components determine which synthetic reactions the system can sustain [1][2].
- cell extracts
- in vitro biosynthesis
- translation machinery
- energy regeneration
References
- So-Jeong Lee; Dong-Myung Kim; Cell-Free Synthesis: Expediting Biomanufacturing of Chemical and Biological Molecules. Molecules 2024, 29, 1878, 10.3390/molecules29081878.
- Srujan Kumar Dondapati; Marlitt Stech; Anne Zemella; Stefan Kubick; Cell-Free Protein Synthesis: A Promising Option for Future Drug Development. BioDrugs 2020, 34, 327-348, 10.1007/s40259-020-00417-y.
- Andrew J. Rice; Tien T. Sword; Kameshwari Chengan; Douglas A. Mitchell; Nigel J. Mouncey; Simon J. Moore; Constance B. Bailey; Cell-free Synthetic Biology for Natural Product Biosynthesis and Discovery. Chem. Soc. Rev. 2025, 54, 4314-4352, 10.1039/d4cs01198h.
- Quentin M. Dudley; Ashty S. Karim; Michael C. Jewett; Cell‐free Metabolic Engineering: Biomanufacturing beyond the Cell. Biotechnol. J. 2014, 10, 69-82, 10.1002/biot.201400330.