Continuous cell-free protein synthesis using glycolytic intermediates as energy sources.
In this work, we demonstrate that glycolytic intermediates can serve as efficient energy sources to regenerate ATP during continuous-exchange cell-free (CECF) protein synthesis reactions. Through the use of an optimal energy source, approximately 10 mg/ml of protein was generated from CECF protein synthesis reaction at greatly reduced reagent costs. Compared with the conventional reactions utilizing phosphoenol pyruvate as an energy source, the described method yields 10-fold higher productivity per unit reagent cost, making the techniques of CECF protein synthesis more realistic alternative for rapid protein production.
Kim
Ho-Cheol
HC
Department of Fine Chemical Engineering and Chemistry, Chungnam National University, Daejeon 305-764, Korea.
Kim
Tae-Wan
TW
Park
Chang-Gil
CG
Oh
In-Seok
IS
Park
Kyungmoon
K
Kim
Dong-Myung
DM
D000255:Adenosine Triphosphate / Q000378:metabolism*; D001674:Bioelectric Energy Sources / Q000382:microbiology*; D004736:Energy-Generating Resources; D004926:Escherichia coli / Q000378:metabolism*; D005947:Glucose / Q000378:metabolism*; D006019:Glycolysis; D014176:Protein Biosynthesis
D000255:Adenosine Triphosphate; D005947:Glucose