Increased recombinant protein production in Escherichia coli strains with overexpressed water-forming NADH oxidase and a deleted ArcA regulatory protein.
Glycolytic flux is increased and acetate production is reduced in Escherichia coli by the expression of heterologous NADH oxidase (NOX) from Streptococcus pneumoniae coupled with the deletion of the arcA gene, which encodes the ArcA regulatory protein. In this study, we examined the overproduction of a model recombinant protein in strains of E. coli expressing NOX with or without an arcA mutation. The presence of NOX or the absence of ArcA reduced acetate by about 50% and increased beta-galactosidase production by 10-20%. The presence of NOX in the arcA strain eliminated acetate production entirely in batch fermentations and resulted in a 120% increase in beta-galactosidase production.
Department of Biological and Agricultural Engineering, Center for Molecular BioEngineering, University of Georgia, Athens, Georgia 30602, USA.
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Vemuri
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Eiteman
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E
Altman
D000085:Acetates / Q000378:metabolism; D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics* / Q000254:growth & development; D017353:Gene Deletion; D015870:Gene Expression / Q000235:genetics; D009247:NADH, NADPH Oxidoreductases / Q000096:biosynthesis* / Q000235:genetics; D010084:Oxidation-Reduction; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics; D012097:Repressor Proteins / Q000235:genetics*; D013296:Streptococcus pneumoniae / Q000201:enzymology / Q000235:genetics*; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics
D000085:Acetates; D001426:Bacterial Proteins; D011994:Recombinant Proteins; D012097:Repressor Proteins; D009247:NADH, NADPH Oxidoreductases; C121789:Nox-2 protein, bacteria; D001616:beta-Galactosidase