Purification and characterization of d-allulose 3-epimerase derived from Arthrobacter globiformis M30, a GRAS microorganism.

An enzyme that catalyzes C-3 epimerization between d -fructose and d -allulose was found in Arthrobacter globiformis strain M30. Arthrobacter species have long been used in the food industry and are well-known for their high degree of safety. The enzyme was purified by ion exchange and hydrophobic interaction chromatographies and characterized as a d -allulose 3-epimerase ( d -AE). The molecular weight of the purified enzyme was estimated to be 128 kDa with four identical subunits. The enzyme showed maximal activity and thermostability in the presence of Mg 2+ . The optimal pH and temperature for enzymatic activity were 7.0–8.0 and 70°C, respectively. The enzyme was immobilized to ion exchange resin whereupon it was stable for longer periods than the free enzyme when stored at below 10°C. In the column reaction, the enzyme activity also maintained stability for more than 4 months. Under these conditions, 215 kg of d -allulose produced per liter immobilized enzyme, and this was the highest production yield of d -allulose reported so far. These highly stable properties suggest that this enzyme represents an ideal candidate for the industrial production of d -allulose.