Journal of agricultural and food chemistryOct 23, 2026

Improving E. coli’s sugar-making process to efficiently produce Lacto-N-tetraose

CRISPR Gene Editing Weekly Brief ↗PubMed ↗DOI ↗

Updated Oct 24, 2026

Abstract

An engineered strain of Escherichia coli achieved 36.29 g/L of Lacto-N-tetraose (LNT) with a 95.6% conversion rate from an intermediate.

Lacto-N-tetraose (LNT) is a crucial component of human milk oligosaccharides.
Current methods for LNT production face challenges due to low conversion efficiency and intermediate accumulation.
A metabolic engineering strategy was successfully applied to improve LNT production in Escherichia coli.
The engineered strain represents the highest conversion efficiency reported for LNT biosynthesis in a single microbial strain.
The integrated approach enhances production efficiency and may reduce downstream purification challenges.

Simplified

Lacto-N-tetraose (LNT), a core component of human milk oligosaccharides (HMOs), plays essential biological roles; however, its production remains challenging due to low conversion efficiency and the accumulation of intermediates. In this study, a systematic metabolic engineering strategy was applied to Escherichia coli BL21(DE3) to enhance LNT production. Key approaches included CRISPR-Cas9-mediated host optimization, chromosomal integration of critical pathway genes, and systematic optimization of fermentation conditions. As a result, the engineered strain achieved an LNT titer of 36.29 g/L, with a molar conversion rate of 95.6% from the intermediate lacto-N-triose II to LNT, representing the highest conversion efficiency reported to date for LNT biosynthesis in a single microbial strain. This integrated strategy significantly improves production efficiency while reducing the burden of downstream purification, providing a robust technological framework for the industrial biosynthesis of LNT and other high-value oligosaccharides.

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Improving E. coli’s sugar-making process to efficiently produce Lacto-N-tetraose

Abstract

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