Use of antioxidants to extend the storage of lyophilized cell-free synthesis system

Modern chemical processes, vital for diverse product manufacturing, often result in substantial energy consumption and environmental waste. As environmental concerns continue to escalate and industries evolve to meet customized demands, biomanufacturing emerges as a promising alternative due to its efficiency, expandability, and eco-friendliness. Cell-free synthesis systems, which harness cellular extracts for biosynthetic reactions, offer a highly adaptable solution for biomanufacturing, particularly when rapid production is required with limited resources. While conventional cell-free systems encounter challenges related to storage and transportation due to the necessity for ultra-cold temperatures, recent studies have demonstrated that these systems can be lyophilized and rehydrated to enable on-demand biomolecule production. Our study aims to enhance the stability of lyophilized cell-free systems. We have discovered the significant role played by antioxidants, specifically dithiothreitol and sodium nitrite, in preserving translational activity during extended storage. This finding represents a significant step forward for decentralized, on-demand protein production using cell-free methods.