A paper on the impact of reducing agents on protein synthesis in PUREfrex® has been published
A paper on the impact of reducing agents on protein synthesis in a reconstituted cell-free protein synthesis system using PUREfrex® by joint research by Gene Frontier, Institute of Science Tokyo has been published in ACS Synthetic Biology.
In this paper, we investigate the impact of reducing agents on protein synthesis in a reconstituted cell-free protein synthesis system (PURE system, Product name: PUREfrex®). Maintaining appropriate redox conditions is essential for protein stability and function. Reducing agents such as dithiothreitol (DTT) and reduced glutathione are commonly used in cell-free reactions to prevent unwanted oxidation.
In this study, we investigated the time-dependent decrease in reducing activity of DTT during prolonged protein synthesis using the PURE system. We identified dissolved oxygen and contaminating metal ions as the primary factors responsible for this loss of activity. Based on these findings, we established a method using a chelator to maintain reducing condition throughout the reaction.
Our findings demonstrate that the reaction conditions can be flexibly optimized not only for the synthesis of proteins with disulfide bonds but also for proteins requiring a reduced state. This versatility provides a more effective strategy for efficient functional protein synthesis, with potential applications in biotechnology and therapeutic protein production.
Impact of Reducing Agents on Protein Synthesis in a Reconstituted Cell-Free Protein Synthesis System
ACS Synthetic Biology (2026), Vol. 15, p1001-1007.
Download paper: https://pubs.acs.org/doi/10.1021/acssynbio.6c00011