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Interaction of Periplasmic Fab Production and Intracellular Redox Balance in <i>Escherichia coli</i> Affects Product Yield

Sophie Vazulka, Matteo Schiavinato, Martin Wagenknecht, Monika Cserjan‐Puschmann, Gerald Striedner

2022ACS Synthetic Biology14 citationsDOIOpen Access PDF

Abstract

gene in the Fab-producing strains. We attributed this to the accumulation of intracellular superoxide, which was measured using flow cytometry. An exogenously supplemented ubiquinone analogue improved Fab yields up to 82%, indicating that partitioning of the quinone pool between aerobic respiration and oxidative folding limited ubiquinone availability and hence disulfide bond formation capacity. Combined, our results provide a more in-depth understanding of the profound effects that periplasmic Fab expression and in particular disulfide bond formation has on the host cell. Thereby, we show new possibilities to elaborate cell engineering and process strategies for improved host cell fitness and process outcome.

Topics & Concepts

DsbAPeriplasmic spaceOxidative foldingBiochemistryEscherichia coliIntracellularProtein disulfide-isomeraseChemistryOxidative phosphorylationRespiratory chainProtein foldingCell biologyBiologyMitochondrionGeneEndoplasmic reticulumBacterial Genetics and BiotechnologyEndoplasmic Reticulum Stress and DiseaseViral Infectious Diseases and Gene Expression in Insects
Interaction of Periplasmic Fab Production and Intracellular Redox Balance in <i>Escherichia coli</i> Affects Product Yield | Litcius