ArticleMicrobial cell factories2025
Enhanced anti-EpCAM ScFv production and reduced acetate overflow in a BW25113-derived E. coli strain with ArcA and Pka deletions.
Article in Microbial cell factories, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
Escherichia coli strains are widely utilized as cell factories for recombinant protein production. However, acetate overflow remains a significant challenge that negatively impacts both biomass yield and protein expression. Here, we evaluated a previously engineered E. coli K 12 BW25113 strain with pka and arcA deletions (RV04) for the expression of a single-chain variable fragments (scFv) derived from 4D5MOC-B, a monoclonal antibody that binds to epithelial cell adhesion molecule (EpCAM) as a biologically important marker for tumor immunotherapy. According to our results, RV04 strain demonstrated a significant growth advantage over both BW25113 and BL21 strains. In minimal M9 medium, RV04 exhibited a maximum cell density that was 44% higher than wild‑type and 11% higher than BL21. In enriched M9 medium, RV04 achieved a remarkable maximum specific growth rate (µ_max) of 0.775 ± 0.003 h⁻¹ and a maximum cell density of 2.1095 ± 0.0205, even under metabolic load. Regarding acetate accumulation, RV04 fully eliminated acetate accumulation within 24 h, whereas BW25113 accumulated acetate up to 0.521 g L
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