Prolonged cell-free protein synthesis using dual energy sources: Combined use of creatine phosphate and glucose for the efficient supply of ATP and retarded accumulation of phosphate.
The accumulation of inorganic phosphate inhibits protein synthesis in cell-free protein synthesis reactions that are energized by high-energy-phosphate-containing compounds. This study developed a new scheme for supplying energy using dual energy sources to enhance the regeneration of ATP and lower the rate of phosphate accumulation. In the proposed scheme, where creatine phosphate (CP) and glucose were simultaneously used as the energy sources, the phosphate released from the CP was subsequently used in the glycolytic pathway for the utilization of the glucose, which enhanced the ATP supply and reduced the rate of inorganic phosphate accumulation. When tested against different proteins, the developed method produced 2-3 times more protein than the conventional ATP regeneration methods using single energy sources.
Kim
Tae-Wan
TW
School of Chemical and Biological Engineering, College of Engineering, Seoul National University, Seoul, Korea.
Oh
In-Seok
IS
Keum
Jung-Won
JW
Kwon
Yong-Chan
YC
Byun
Ju-Young
JY
Lee
Kyung-Ho
KH
Choi
Cha-Yong
CY
Kim
Dong-Myung
DM
D000255:Adenosine Triphosphate / Q000378:metabolism*; D002474:Cell-Free System / Q000378:metabolism*; D004735:Energy Transfer / Q000502:physiology; D004926:Escherichia coli / Q000502:physiology*; D029968:Escherichia coli Proteins / Q000096:biosynthesis*; D005947:Glucose / Q000378:metabolism*; D010725:Phosphocreatine / Q000378:metabolism*; D014176:Protein Biosynthesis / Q000502:physiology*
D029968:Escherichia coli Proteins; D010725:Phosphocreatine; D000255:Adenosine Triphosphate; D005947:Glucose