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KMID : 0380020040190060467
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Korean Journal of Biotechnology and Bioengineering
2004 Volume.19 No. 6 p.467 ~ p.470
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Regeneration of ATP through an Activated Glycolytic Pathway in a Cell-free Extract and its Application for Protein Expression
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Kim Dong-Myung
Keum Jung-Won
Kim Tae-Wan
Oh In-Seok
Choi Cha-Yong
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Abstract
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We have investigated the key parameters affecting ATP regeneration in a cell-free protein synthesis system derived from Escherichia coli. When glucose-6-phosphate was used as an energy source, the efficiency of ATP regeneration sharply responded to pH change of reaction mixture. In addition, both productivity and reproducibility of protein synthesis was substantially enhanced by introducing appropriate amount of NAD into the reaction mixture. As a result, through the activation of glycolytic pathway under an optimal pH, the batch cell-free system produced over of protein in a 1 mL reaction.
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