Enzymatic Synthesis of Oligosaccharides

Enzymatic synthesis of oligosaccharides involves hydrolysis of polysaccharides or synthesis from smaller sugars using glycosyltransferases or glycosidases. Glycosyltransferases catalyze the transfer of monosaccharides from sugar nucleotides to sugar acceptors. In addition to hydrolase activity, glycosidase also has transferase activity. In fact, the hydrolysis of glycosidic bonds is a special case of transglycosylation where the acceptor is water. Therefore, oligosaccharides can be produced by enzymatic synthesis using disaccharides as substrates with glycosidases as catalysts. Glycosidases are more abundant in nature.

Fig.1 Overview of enzymatic processes for conversion of lactose to oligosaccharides. (Gänzle, 2012)

Enzymatic Synthesis for Oligosaccharide Manufacture at CD BioGlyco

Enzymatic cascade reactions are powerful technologies to develop environmentally friendly and cost-effective synthetic processes to Manufacture Oligosaccharides. We provide high-quality enzymatic cascade synthesis technology for oligosaccharide production. Take enzymatic cascade synthesis of lacto-N-biose-type (LNT) and lacto-N-neo-type (LNnT) human milk oligosaccharides as an example, our scheme is as follows.

• Production of recombinant enzymes and enzyme activity assay
• Synthesis of LNnT oligomers (mainly GlcNAc terminated)

The synthesis of LNnT oligomers is carried out at a suitable temperature and time. The reaction is buffered with a neutral buffer containing chloride salts, UDP-Gal, DP-GlcNAc, and lactose. Then add β1,4-galactosyltransferase-1, β1,3-galactosyltransferase, and alkaline phosphatase to start the reaction. Heat treatment is used to denature the enzyme to stop the reaction. The reaction is then centrifuged and the precipitated enzyme is removed.

• Synthesis of LNnT oligomers (Gal terminated)

Synthesis of β1,4-galactosyltransferase-1-terminated LNnT oligomers requires treatment of GlcNAc-terminated oligomers with low-concentration lactose solution. After heat treatment to stop the reaction, add β1,4-galactosyltransferase-1, alkaline phosphatase, and UDP-Gal to incubate at a certain temperature.

• Synthesis of LNT oligomers (Gal terminated)

The method of synthesizing LNnT oligomer terminated with β1,3-galactosyltransferase is similar to the above. A low-concentration lactose solution is used to treat terminal GlcNAc oligomers as described. After heat treatment to stop the reaction, add β1,3-galactosyltransferase, alkaline phosphatase, and UDP-Gal and incubate at a certain temperature.

Fig.2 Enzymatic cascade synthesis of LNT and LNnT. (CD BioGlyco)

Applications

• Enzymatic synthesis is used to produce products with well-defined oligosaccharides.
• Enzymatic synthesis is used as a complementary technique for the synthesis of human milk oligosaccharides, a class of oligosaccharides with important health benefits.
• The stereoselective and regioselective properties of enzyme catalysis are important advantages for oligosaccharide production and are complementary tools to chemical methods.

Advantages

• Enzymatically synthesized oligosaccharides provide well-defined products or product distributions.
• Enzymatic synthesis is an environmentally friendly synthesis method.
• The technology of enzymatic synthesis of oligosaccharides has been constantly breaking through.

CD BioGlyco provides complete One-Stop Solutions for Carbohydrate Manufacture, and our experts customize experimental schemes according to clients' scientific research projects. Our dedicated team of experts works tirelessly to provide Enzymatic Process for Oligosaccharide Manufacture that meets clients' needs and requirements. We offer comprehensive and advanced enzymatic synthesis, especially enzymatic cascade synthesis technology for oligosaccharide production. We're here to help and don't hesitate to contact us for more information.

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• Enzymatic Isomerization for Oligosaccharide Manufacture
• Enzymatic Epimerization for Oligosaccharide Manufacture