Harnessing 3-Fucosyllactose: Dynamic Regulation Unlocks Efficient l-fucose Production in Escherichia coli

Area of Science:

• Biotechnology
• Metabolic Engineering
• Microbial Synthesis

Background:

• L-fucose is vital for infant intestinal health and immune development, found in human milk oligosaccharides.
• It is widely utilized in infant formula, functional foods, and nutritional supplements.
• Efficient and sustainable production methods are needed.

Purpose of the Study:

• To develop a novel synthetic route for efficient and sustainable l-fucose biosynthesis.
• To engineer Escherichia coli for high-titer l-fucose production.
• To establish a safe, plasmid-free, and antibiotic-free production system.

Main Methods:

• Engineered Escherichia coli BL21star (DE3) as the host.
• Introduced α-1,3-fucosyltransferase (FutM2) and α-l-fucosidase (BiafcB).
• Knocked out competing pathways, balanced enzyme expression, and employed dynamic metabolic control with a temperature-coupled mixed carbon source strategy.

Main Results:

• Achieved a high l-fucose titer of 86.22 g/L in an 89-hour fed-batch fermentation.
• Developed a stable, plasmid-free, and antibiotic-free production system.
• Demonstrated efficient metabolic flux redirection towards l-fucose synthesis.

Conclusions:

• The engineered Escherichia coli system enables efficient and sustainable l-fucose production.
• This novel approach offers a safe and scalable method for industrial l-fucose manufacturing.
• The developed system has significant implications for the food and nutritional supplement industries.