Recombinant escherichia coli for efficiently preparing high-purity sialyllactose-n-tetrasaccharide a, and preparation method and application thereof
By integrating a specific glycosyltransferase gene into Escherichia coli BL21(DE3) and optimizing the metabolic pathway, a recombinant Escherichia coli strain capable of efficiently producing high-purity sialyl lactose-N-tetrasaccharide a was constructed, solving the problems of insufficient yield and purity in existing technologies and realizing efficient and low-cost industrial production.
Patent Information
- Application Number
- CN202610012964.0
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2026-01-07
- Publication Date
- 2026-05-01
- Estimated Expiration
- 2046-01-07
AI Technical Summary
In existing technologies, the natural content of sialyl lactose-N-tetrasaccharide a in breast milk is extremely low. Traditional chemical synthesis is cumbersome and costly, enzymatic synthesis is inefficient, and engineered strains have low catalytic activity, making it difficult to meet industrial requirements in terms of yield and purity of sialyl lactose-N-tetrasaccharide a.
Recombinant Escherichia coli BL21(DE3) was used as the chassis host. The genes of β-1,3-N-acetylglucosamine transferase, β-1,3-galactosyltransferase and α-2,3-sialic acid glycosyltransferase were integrated and gene editing was performed using λ-Red homologous recombination technology to optimize metabolic pathways and construct a recombinant Escherichia coli that can efficiently produce high-purity sialyllactose-N-tetrasaccharide a. Glycerol and lactose were used as carbon sources during fermentation.
It significantly improved the yield and purity of sialyl lactose-N-tetrasaccharide a, enabling efficient and low-cost industrial production, reducing the generation of non-target byproducts, and showing good prospects for industrial application.