Method for producing seaweed-derived agarotriose and its use as a prebiotic

Prebiotic manufacturing method for obtaining high-purity agarotriose through sequential reactions of two enzymes derived from gut microbiota

This technology is a manufacturing method that obtains agarotriose by first degrading agar, agarose, or neoagarohexaose with beta-agarase, then reacting the resulting product with neoagarobiose hydrolase, and purifying it using a size-exclusion column. Both enzymes are derived from the gut bacterium Bacteroides plebeius strain DSM 17135.

Research on the prebiotic effects of oligosaccharides derived from red algae has historically been conducted using mixtures rather than purified standard substances, making it impossible to identify which specific components effectively alter the gut microbiome. In existing 3,6-anhydro-L-galactose production processes, agarotriose is generated as a byproduct that requires further enzymatic degradation, and specific process technologies for obtaining pure agarotriose have not been previously established domestically.

This technology enables the stepwise degradation of red algae polysaccharides using only enzymatic reactions, without the need for acid or buffer-based substrate pretreatment, and recovers agarotriose with high yield through size-exclusion purification that minimizes loss. The agarotriose obtained in this way possesses prebiotic properties that are selectively metabolized by probiotic microorganisms such as Bacteroides and Bifidobacterium. It can be applied to the production of gut health prebiotic food ingredients, anti-inflammatory functional materials, and standard substances for gut microbiome research, allowing for the industrial-scale acquisition of single-component functional oligosaccharides from seaweed polysaccharides.

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Key Features:

• A step of reacting a substrate, which is any one of agar, agarose, or neoagarohexaose, with the beta-agarase of SEQ ID NO: 1
• A step of reacting the beta-agarase reaction product with the neoagarobiose hydrolase of SEQ ID NO: 2 or 6
• A step of separating the target product, agarotriose, from the completed enzymatic reaction mixture using a size-exclusion column to purify it to a high degree
• A beta-agarase derived from the Bacteroides plebeius DSM 17135 strain that degrades the substrate into neoagarotetraose and neoagarobiose

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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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