It is useful for protein pharmaceuticals, e. g. recombinant therapeutic proteins, to synthesize large amount of any desired protein from DNA and mRNA. Cell-free protein synthesis system was developed as a model to study the mechanism of translation and succeeded to provide a lot of proteins even if its property suppressed cell survival. For protein pharmaceuticals, protein folding is an important point. In recombinant DNA production system, newly synthesized proteins pass through the chaperone system of E. coli. In cell-free protein synthesis, some key factors of chaperone are added for the improvement of protein folding but the instability of chaperone affects the amount of folded protein. Here we report that the effect of amines, alcohols, carboxylic acid and thiols to translation and protein folding by the using of cell-free protein synthesis system and we explored newly protein folding molecules. To estimate translation and protein folding efficiency, glutathione S-transferase (GST) was synthesized and the amount of GST and the activity of GST were measured. The addition of thiols, the amount of synthesized GST was increased and in the case of monoamines, the amount of synthesized GST was decreased but no significant difference in the activity of GST was not observed. By the addition of diamines, both the amount of GST and the activity of GST were decreased. This suppression did not alter by the amount of mRNA. To analyze the effect of the conformation of diamine, we testified cis-1,2- diaminocyclohexane, trans-1,2-diaminocyclohexane and diaminobenzene. It is revealed that the conformation of diamine is an important factor of the translation and folding efficiency.
