Metabolic engineering of Escherichia coli strains for the in vivo phosphorylase-mediated synthesis of disaccharides
Abstract
BackgroundEnzymatic synthesis of rare disaccharides by reverse-phosphorolysis is a potentially sustainable route to produce high-value glycosides for human health and nutrition. In vivo synthesis using microorganisms expressing recombinant glycoside phosphorylases is an attractive strategy, as it may overcome key limitations associated with conventional chemical and enzymatic approaches. However, successful implementation requires dedicated metabolic chassis enabling efficient precursor supply and carbon flux redirection. In Escherichia coli, inactivation of the phosphotransferase system (PTS) combined with deletion of pfkA constitutes a promising strategy to decouple sugar uptake from their concomitant phosphorylation, promote the accumulation of key sugar phosphates, and enable reverse biosynthetic pathways for oligosaccharide production.
ResultsWe first established β-1,2-mannobiose production as a proof of concept. The expression of heterologous permease GalP restored the growth on mannose in a PTS⁻ background and allowed mannose into the reverse biosynthetic
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BibTeX
@article{tedesco2026metabolic,
title = {{Metabolic engineering of Escherichia coli strains for the in vivo phosphorylase-mediated synthesis of disaccharides}},
author = {Pietro Tedesco and Julien Durand and Laurence Tarquis and Gabrielle Potocki-Veronese and Fabien Létisse},
journal = {Microbial Cell Factories},
year = {2026},
month = jul,
doi = {10.1186/s12934-026-03051-5},
url = {https://hal.science/hal-05761484v1},
note = {CC BY-NC-ND 4.0},
}
RIS
TY - JOUR
TI - Metabolic engineering of Escherichia coli strains for the in vivo phosphorylase-mediated synthesis of disaccharides
AU - Pietro Tedesco
AU - Julien Durand
AU - Laurence Tarquis
AU - Gabrielle Potocki-Veronese
AU - Fabien Létisse
PY - 2026
DA - 2026/07/07
JO - Microbial Cell Factories
DO - 10.1186/s12934-026-03051-5
UR - https://hal.science/hal-05761484v1
AB - BackgroundEnzymatic synthesis of rare disaccharides by reverse-phosphorolysis is a potentially sustainable route to produce high-value glycosides for human health and nutrition. In vivo synthesis using microorganisms expressing recombinant glycoside phosphorylases is an attractive strategy, as it may overcome key limitations associated with conventional chemical and enzymatic approaches. However, successful implementation requires dedicated metabolic chassis enabling efficient precursor supply and carbon flux redirection. In Escherichia coli, inactivation of the phosphotransferase system (PTS) combined with deletion of pfkA constitutes a promising strategy to decouple sugar uptake from their concomitant phosphorylation, promote the accumulation of key sugar phosphates, and enable reverse biosynthetic pathways for oligosaccharide production. ResultsWe first established β-1,2-mannobiose production as a proof of concept. The expression of heterologous permease GalP restored the growth on mannose in a PTS⁻ background and allowed mannose into the reverse biosynthetic
SN - 1475-2859
ER -