Cell-free Systems: Recent Advances and Future Outlook
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pISSN 1226-8372 eISSN 1976-3816
REVIEW PAPER
Cell-free Systems: Recent Advances and Future Outlook Jamin Koo, Jiho Yang, and Hyunjun Park
Received: 12 January 2020 / Revised: 6 May 2020 / Accepted: 8 May 2020 © The Korean Society for Biotechnology and Bioengineering and Springer 2020
Abstract Cell-free systems utilize a subset of cellular components without intact cell wall and/or membranes. The system was first invented for use in fermentation. Subsequent improvements enabled its application in protein synthesis, which is still the most common use of the system. Lately, attempts have been reported where metabolic engineering concepts and techniques were applied to cell-free systems and/or vice-versa. These attempts and advances led to exciting discoveries about biochemical reactions, as well as properties and/or structures of cellular components that make up complex biological systems. This review will first provide a basic overview and brief history of the cell-free system. Then, explanation on recent advances in the field will be provided, followed by notes on the innovative applications. Future outlook of the field will also be covered with the emphasis on how the emerging data science methods can be applied to improve the system and its applicability. Keywords: cell-free, protein synthesis, metabolic engineering, artificial intelligence, unnatural amino acids
1. Introduction Cell-free (CF) systems refer to biochemical systems without intact cells. CF systems utilize parts of cellular components such as nucleotides, metabolites, and proteins that reside on and/or inside cellular membranes. Two approaches have been developed for obtaining these components—topdown and bottom-up [1]. The former begins with breaking the cell wall and/or membranes and removing undesirable Jamin Koo*, Jiho Yang, Hyunjun Park Department of Chemical Engineering, Hongik University, Seoul 04066, Korea Tel: +82-2-320-3006 E-mail: [email protected]
components. Only few purification steps are conducted in the process. In contrast, the latter begins with purifying the desired subcellular components individually and combining them afterward. During the combination step, concentrations of the purified components can be adjusted, resulting in greater freedom for designing a CF system than when using the top-down approach. CF system was first reported in 1897 [2]. Buchner used the system for alcoholic fermentation, but no following work was reported for the next 50 years. In 1961, a report came out that described using the system for polypeptide synthesis [3]. Most of the subsequent reports were about developing and/or applying the CF for protein synthesis, which still remains to be the most common use of the system. Since the mid 2000s, researchers began to apply metabolic engineering to CF systems, mostly to enhance production of metabolites. EEHP (ethyl(S)-2-ethoxy-3-(pmethoxy-phenyl)propanoate) is one of the key chemicals used in synthesizing therapeutic molecules for diabetes. The productivity of its production process increased n
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