D-(-)-Fructose → Psicose

The activity of the ketose 3-epimerase enzyme before purification may be measured by measuring the amount of D-psicose produced with a D-fructose substrate. Specifically, an enzyme reaction mixture composition containing 50 mM tris-HCl buffer (pH 7.0; 200 ml), 0.2 M D-fructose (375 μl), enzyme liquid (100 μl), and 10 mM manganese chloride (75 μl) is allowed to react at 55° C. for 30 min, and the reaction is quenched by boiling the mixture for 2 min. The resulting liquid composition is then measured by HPLC. One unit of enzyme activity is defined as the amount of enzyme that epimerizes D-fructose and produces 1 μmol of D-psicose per minute under the foregoing conditions. The enzyme unit for the epimerization of D-psicose to D-fructose in the reverse reaction is defined in the same manner through measurement performed under the same conditions.

1. customSpecifically, an enzyme reaction mixture composition
2. customto react at 55° C. for 30 min
3. customthe reaction is quenched
4. customfor 2 min

ORD record

ord-58c092e56ebf480180bb3c191e8b7d7c

Source dataset

uspto-grants-2015_06

DOI

10.6084/m9.figshare.5104873.v1

Patent number

US09057062B2