Characterization of a d-tagatose 3-epimerase from Caballeronia fortuita and its application in rare sugar production

Abstract Recently, rare sugars have caused extensively attention due to their beneficial physiological functions and potential applications in food systems and medical fields. Ketose 3-epimerase (KEase) can catalyze reversibly the epimerization between ketoses which is the pivotal enzyme in Izumoring strategy and an effective tool for biological production of rare sugars. In this work, a KEase from Caballeronia fortuita was recombined and characterized as a d -tagatose 3-epimerase (DTEase, EC 5.1.3.31). The recombinant DTEase displayed the highest activity at pH 7.5 and 65 °C in the presence of Co 2+ . The recombinant DTEase displayed the relatively high thermostability and the half-life ( t 1/2 ) was determined to be 7.13, 5.13, and 1.05 h at 50, 55, and 60 °C, respectively. The recombinant DTEase had a wide substrate specificity and the specific activities towards d -tagatose, d -allulose, d -fructose and l -sorbose were measured to be 801 ± 2.3, 450 ± 2.7, 270 ± 1.5 and 55 ± 1.8 U mg −1 , respectively. So far, the recombinant DTEase exhibited the highest specific activity towards d -tagatose compared with other reported KEases. Furthermore, the recombinant DTEase could produce 314.2 g/L d -sorbose from 500 g/L d -tagatose and 147.0 g/L d -allulose from 500 g/L d -fructose, with a transformation ratio of 68.2% and 29.4%, respectively. The recombinant DTEase could realize effectively the transformations between various ketoses and was a prominent candidate for production of rare sugars.