Recombinant escherichia coli for producing lacto-n-triose, and two-strain-coupled fermentation method based on same

By expressing a specific enzyme in recombinant Escherichia coli and coupling it with yeast cells for fermentation, the problem of efficient production of lactose-N-trisaccharide was solved, achieving high yield and high conversion rate, and promoting its application in functional foods.

WO2026138534A1PCT designated stage Publication Date: 2026-07-02JIANGNAN UNIV
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Patent Information

Authority / Receiving Office
WO · WO
Patent Type
Applications
Current Assignee / Owner
JIANGNAN UNIV
Filing Date
2025-12-12
Publication Date
2026-07-02

AI Technical Summary

Technical Problem

Existing technologies make it difficult to produce lactose-N-trisaccharide efficiently and on a large scale. Furthermore, the extraction methods for natural LNT II in breast milk are complex and costly, and the synthetic routes are cumbersome, which limits its application in functional foods.

Method used

Recombinant Escherichia coli was used to express N-acetylglucosamine 1-kinase, uridine diphosphate-N-acetylglucosamine pyrophosphorylase and β-1,3-N-acetylglucosamine aminotransferase, and combined with yeast cell-coupled fermentation to synthesize lactose-N-trisaccharide using N-acetylglucosamine and lactose as substrates.

Benefits of technology

High yield (52.19 g/L) and high conversion rate of lactose-N-trisaccharide were achieved, simplifying the synthesis route and showing good industrialization prospects.

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Abstract

The present invention belongs to the technical field of biology and relates to a recombinant Escherichia coli for producing lacto-N-triose and a two-strain-coupled fermentation method based on same. The provided recombinant Escherichia coli for producing lacto-N-triose expresses three genes N-acetylhexosamine 1-kinase gene Nahk, uridine diphosphate-N-Acetylglucosamine pyrophosphorylase gene GlmU, and β-1,3-N-acetylglucosaminyltransferase gene LgtA. The coupled catalysis of the obtained recombinant Escherichia coli and yeast to produce lacto-N-triose is a two-stage fermentation process, wherein the first stage is to culture engineered Escherichia coli, and the second stage is to perform coupled fermentation by means of the yeast and the recombinant Escherichia coli, the yield on a 5L fermentation tank being 52.2 g / L. The provided fermentation production process is simple and has a high conversion rate, and thus has good industrialization prospects.
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