D-(+)-Cellobiose (CNP0226604.15) → Epilactose

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Through the specification of the present invention, the activity of the cellobiose 2-epimerase was determined as a lactose 2-epimerase activity, which catalyzes epimerization of lactose to form epilactose, using lactose as the substrate obtainable in highly purified form at low cost. The lactose 2-epimerase activity was measured as follows. To 1,000 μl of substrate solution, which contains lactose to give a final concentration of 35.1 mM and acetate buffer (pH 6.0) to give a final concentration of 20 mM of the reaction solution, 200 μl of a enzyme solution was added to give a 1,200 μl of the reaction solution, followed by reaction at 50° C. for 20 minutes. The reaction was stopped by heating for 10 minutes in a boiling water bath. Then reaction mixture was subjected to HPLC chromatography to determine the amount of epilactose produced by the reaction. The HPLC chromatography was conducted at the following conditions;

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