Abstract
AbstractOverexpression of gluconeogenic phosphoenolpyruvate carboxykinase (PCK) under glycolytic conditions enables Escherichia coli to maintain a greater intracellular ATP concentration and, consequently, to up-regulate genes for amino acid and nucleotide biosynthesis. To investigate the effect of a high intracellular ATP concentration on heterologous protein synthesis, we studied the expression of a foreign gene product, enhanced green fluorescence protein (eGFP), under control of the T7 promoter in E. coli BL21(DE3) strain overexpressing PCK. This strain was able to maintain twice as much intracellular ATP and to express two times more foreign protein than the control strain. These results indicate that a high energy-charged cell can be beneficial as a protein-synthesizing cell factory. The potential uses of such a cell factory are discussed.
Highlights
YZG Overexpression of gluconeogenic phosphoenolpyruvate carboxykinase (PCK) Y[G under glycolytic conditions enables Escherichia coli to maintain a greater Y\G intracellular Adenosine triphosphate (ATP) concentration and, to up-regulate genes for amino Y]G acid and nucleotide biosynthesis
To investigate the effect of a high intracellular Y^G ATP concentration on heterologous protein synthesis, we studied the expression of Y_G a foreign gene product, enhanced green fluorescence protein, under control YG of the T7 promoter in E. coli BL21(DE3) strain overexpressing PCK
This strain ZWG was able to maintain twice as much intracellular ATP and to express two times ZXG more foreign protein than the control strain. These results indicate that a high ZYG energy-charged cell can be beneficial as a protein-synthesizing cell factory
Summary
YZG Overexpression of gluconeogenic phosphoenolpyruvate carboxykinase (PCK) Y[G under glycolytic conditions enables Escherichia coli to maintain a greater Y\G intracellular ATP concentration and, to up-regulate genes for amino Y]G acid and nucleotide biosynthesis. To investigate the effect of a high intracellular Y^G ATP concentration on heterologous protein synthesis, we studied the expression of Y_G a foreign gene product, enhanced green fluorescence protein (eGFP), under control YG of the T7 promoter in E. coli BL21(DE3) strain overexpressing PCK. ]`G In this paper, we artificially expressed heterologous enhanced green fluorescence ^WG protein (eGFP) in E. coli BL21(DE3) that maintain a high intracellular ATP to ^XG determine whether this strain is eligible for use as a powerful, protein-synthesizing cell ^YG factory. ^ZG Results ^[G To estimate heterologous protein expression in a high energy-charged cell, enhanced ^\G green fluorescence protein (eGFP) was expressed under control of the T7 promoter in ^]G an E. coli BL21(DE3) host (Table 1).
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