Methods for producing diverse and novel fucosyl oligosaccharides and their applications

Novel prebiotic oligosaccharides produced by fucosylating various sugars using alpha-1,2-fucosyltransferase

This technology involves a method for producing various fucosyl oligosaccharides by reacting a GDP-L-fucose donor with sugar acceptors such as glucose, galactose, cellobiose, xylose, and agarobiose using alpha-1,2-fucosyltransferase. It also includes prebiotic pharmaceutical, cosmetic, and food compositions containing these oligosaccharides. The method covers both enzymatic reactions and direct production using recombinant E. coli or yeast engineered with the GDP-L-fucose biosynthetic pathway.

Fucosyl oligosaccharides are key components of human milk oligosaccharides (HMOs) that are selectively fermented by beneficial gut bacteria, inhibit pathogen colonization, and have been reported to enhance memory and promote nerve cell growth. However, the limited variety of available fucosyl oligosaccharides has hindered the ability to efficiently create diverse structures and verify their prebiotic properties.

This technology enables the creation of novel structures such as fucosylglucose, fucosylxylose, and fucosylagarobiose by selecting one of three enzymes with different amino acid sequences to attach fucose to mono- and disaccharides. It also facilitates mass production through fed-batch cultivation of recombinant strains. The resulting oligosaccharides promote the growth of beneficial bacteria like Bifidobacterium and Lactobacillus. These can be applied as HMO substitutes in infant formula, gut health pharmaceuticals, and microbiome cosmetics, allowing for the industrial-scale supply of various fucosyl sugars with functions similar to those found in human milk.

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Key Features:

• Prebiotic pharmaceutical composition comprising a fucosyl oligosaccharide selected from 9 types, including 2'-fucosylglucose, 2'-fucosylgalactose, 2'-fucosylcellobiose, and 2'-fucosylagarobiose
• Prebiotic composition that promotes the growth of beneficial gut bacteria such as Bifidobacterium, Lactobacillus, or Streptococcus species and inhibits the growth of harmful bacteria
• Step of producing fucosyl oligosaccharides by reacting a sugar acceptor and a GDP-L-fucose donor with an alpha-1,2-fucosyltransferase of any one of SEQ ID NOs: 1 to 3
• Step of culturing recombinant E. coli or yeast introduced with ManB, ManC, Gmd, and WcaG expression vectors and an alpha-1,2-fucosyltransferase expression vector in the presence of a sugar acceptor and glycerol

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This invention was developed with support from the National Research Foundation of Korea for the establishment of a biotechnological production platform for fucose for food and medical materials.

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