Regeneration of adenosine triphosphate from glycolytic intermediates for cell-free protein synthesis.
A new approach for adenosine triphosphate (ATP) regeneration in a cell-free protein synthesis system is described. We first show that pyruvate can be used as a secondary energy source to replace or supplement the conventional secondary energy source, phosphoenol pyruvate (PEP). We also report that glucose-6-phosphate, an earlier intermediate of the glycolytic pathway, can be used for ATP regeneration. These new methods provide more stable maintenance of ATP concentration during protein synthesis. Because pyruvate and glucose-6-phosphate are the first and last intermediates of the glycolytic pathway, respectively, the results also suggest the possibility of using any glycolytic intermediate, or even glucose, for ATP regeneration in a cell-free protein synthesis system. As a result, the methods described provide cell-free protein synthesis with greater flexibility and cost efficiency.
Kim
D M
DM
Department of Chemical Engineering, Stanford University, Stanford, California 94305-5025, USA.
Swartz
J R
JR
D000255:Adenosine Triphosphate / Q000096:biosynthesis* / Q000378:metabolism; D000596:Amino Acids / Q000378:metabolism; D002474:Cell-Free System; D004734:Energy Metabolism; D019298:Glucose-6-Phosphate / Q000378:metabolism; D006019:Glycolysis; D007700:Kinetics; D010728:Phosphoenolpyruvate / Q000378:metabolism; D014176:Protein Biosynthesis; D019289:Pyruvic Acid / Q000378:metabolism; D013997:Time Factors
D000596:Amino Acids; D019298:Glucose-6-Phosphate; D010728:Phosphoenolpyruvate; D019289:Pyruvic Acid; D000255:Adenosine Triphosphate