Bacterial cell-free expression technology to in vitro systems engineering and optimization

• Main Author: Filippo Caschera
• Format: Article
• Language: English
• Published: KeAi Communications Co., Ltd. 2017-06-01
• Series: Synthetic and Systems Biotechnology
• Subjects: Cell-free protein synthesis; Engineering; Optimization; Polysaccharide; ATP-regeneration; Machine-learning; Synthetic minimal cell
• Online Access: http://www.sciencedirect.com/science/article/pii/S2405805X1730056X

Cell-free expression system is a technology for the synthesis of proteins in vitro. The system is a platform for several bioengineering projects, e.g. cell-free metabolic engineering, evolutionary design of experiments, and synthetic minimal cell construction. Bacterial cell-free protein synthesis system (CFPS) is a robust tool for synthetic biology. The bacteria lysate, the DNA, and the energy module, which are the three optimized sub-systems for in vitro protein synthesis, compose the integrated system. Currently, an optimized E. coli cell-free expression system can produce up to ∼2.3 mg/mL of a fluorescent reporter protein. Herein, I will describe the features of ATP-regeneration systems for in vitro protein synthesis, and I will present a machine-learning experiment for optimizing the protein yield of E. coli cell-free protein synthesis systems. Moreover, I will introduce experiments on the synthesis of a minimal cell using liposomes as dynamic containers, and E. coli cell-free expression system as biochemical platform for metabolism and gene expression. CFPS can be further integrated with other technologies for novel applications in environmental, medical and material science.