title: Expression of an anaplerotic enzyme, pyruvate carboxylase, improves recombinant protein production in Escherichia coli.
abstract: Anaplerotic enzyme reactions are those which replenish tricarboxylic acid intermediates that are withdrawn for the synthesis of biomass. In this study, we examined recombinant protein production in Escherichia coli containing activity in an additional anaplerotic enzyme, pyruvate carboxylase. In batch fermentations, the presence of pyruvate carboxylase resulted in 68% greater production of the model protein, beta-galactosidase, 41% greater cell yield, and 57% lower acetate concentration. We discuss why these results indicate that acetate concentration does not limit cell growth and protein synthesis, as predicted by other researchers, and suggest instead that the rate of acetate formation represents an inefficient consumption of glucose carbon, which is reduced by the presence of pyruvate carboxylase.
authors.affiliation: Center for Molecular BioEngineering, Department of Biological and Agricultural Engineering, University of Georgia, Athens, Georgia 30602, USA.
authors.forename: J C
authors.initials: JC
authors.lastname: March
authors.forename: M A
authors.initials: MA
authors.lastname: Eiteman
authors.forename: E
authors.initials: E
authors.lastname: Altman
mesh_terms: D001709:Biotechnology / Q000379:methods; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D011766:Pyruvate Carboxylase / Q000096:biosynthesis / Q000502:physiology*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D001616:beta-Galactosidase / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D011994:Recombinant Proteins; D001616:beta-Galactosidase; D011766:Pyruvate Carboxylase