ArticleMicrobial cell factories2026
Mechanism-guided control of intracellular alkalinization and process pH for production of recombinant human B-type natriuretic peptide under the cSAT scheme in Escherichia coli.
Article in Microbial cell factories, 2026. 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
backgroundTag-based high-load expression in Escherichia coli can be accompanied by substantial drift of broth and intracellular pH, but mechanistic links between intracellular alkalinization, metabolism and process levers remain unclear.
resultsUsing a cleavable self-aggregating tag (cSAT) to produce recombinant human B-type natriuretic peptide (rhBNP), we combine broth pH tracking, AF-C ratiometric intracellular pH imaging and multi-omics, which together suggest that ammoniagenic amino-acid catabolism, glyoxylate-centred carbon rerouting and respiratory shifts are major contributors to intracellular alkalinization. Guided by this proton-economy model, stepwise medium engineering (complex nitrogen, phosphate, glucose and ammonium sulfate with a tuned carbon to nitrogen ratio) lowers shake-flask broth pH from values above 8.5 to about 7.0, increases titers 2.9-fold over LB medium to 115.4 mg/L and, in 3-L fed-batch, yields 662.1 mg/L rhBNP while maintaining biomass.
conclusionsProton-economy-based process design stabilizes pH and productivity during production of rhBNP under the cSAT scheme in E. coli.
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