Oxalate enhances protein synthesis in cell-free synthesis system utilizing 3-phosphoglycerate as energy source.
Herein, we report our results showing that the productivity of cell-free protein synthesis can be enhanced through the regulation of the in vitro metabolism of an energy source. In a reaction mixture utilizing 3-phosphoglycerate (3PG) as an energy source, the supply of ATP was significantly enhanced when the reaction mixture was supplied with sodium oxalate, a potent inhibitor of phosphoenolpyruvate synthetase (PPS). The productivity of protein synthesis was also increased by approximately 70% upon the addition of oxalate. It was presumed that this enhancement in ATP supply resulted from the prevention of the pyruvate --> PEP reaction, which causes nonproductive ATP consumption. For the initial presence of 2.1 mM sodium oxalate, approximately 720 microg/ml chloramphenicol acetyltransferase (CAT) was produced after 3 h of incubation at 37 degrees C.
Kuem
Jung-Won
JW
School of Chemical and Biological Engineering, Seoul National University, Seoul 151-742, Korea.
Kim
Tae-Wan
TW
Park
Chang-Gil
CG
Choi
Cha-Yong
CY
Kim
Dong-Myung
DM
D000255:Adenosine Triphosphate / Q000378:metabolism; D002474:Cell-Free System / Q000187:drug effects / Q000378:metabolism*; D015500:Chloramphenicol O-Acetyltransferase / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005988:Glyceric Acids / Q000378:metabolism*; D010070:Oxalates / Q000494:pharmacology*; D010728:Phosphoenolpyruvate / Q000378:metabolism; D017866:Phosphotransferases (Paired Acceptors) / Q000037:antagonists & inhibitors / Q000378:metabolism; D014176:Protein Biosynthesis / Q000187:drug effects*
D004791:Enzyme Inhibitors; D005988:Glyceric Acids; D010070:Oxalates; D010728:Phosphoenolpyruvate; C005156:3-phosphoglycerate; D000255:Adenosine Triphosphate; D015500:Chloramphenicol O-Acetyltransferase; D017866:Phosphotransferases (Paired Acceptors); C022509:pyruvate, water dikinase