| 초록 |
The unique flexibility and throughput of cell-free protein synthesis have made this technology a promising alternative to cell-based protein production methods. However, the limitations associated with energy sources have prevented its widespread applications. Commonly used creatine phosphate causes early cessation of protein synthesis due to the depletion of Mg2+. Although glucose can be used as an energy source, the use of glucose has the problem of proton accumulation with is also inhibitory for protein synthesis. In this study, we attempted to devise a ‘clean’ method for ATP regeneration that does not leave any by-products that interfere with protein synthesis reaction. In place of previously used energy sources, we added formate to the mixture along with formate dehydrogenase. Since the conversion of formate into CO2 accompanies the regeneration of NADH, we expected that the regenerated NADH would be used for regeneration of ATP through the oxidative phosphorylation reaction in the reaction mixture for CFPS. |
