D-Mannose isomerase can reversibly catalyze D-fructose to D-mannose which has various beneficial effects. A novel D-mannose isomerase gene (PsMIaseA) from Pseudomonas syringae was cloned and expressed in Escherichia coli. The recombinant D-mannose isomerase (PsMIaseA) showed the highest amino acid sequence homogeneity of 50% with ManI from Thermobifda fusca. PsMIaseA was purified through Ni-NTA chromatography, and its specific activity was 818.6 U mg-1. The optimal pH and temperature of PsMIaseA were pH 7.5 and 45 °C, respectively. The enzyme was stable within a wide pH range from 5.0 to 10.0. It could efficiently convert D-fructose to D-mannose without any metal ions. When PsMIaseA was incubated with 600 g/L D-fructose for 6 h, the space-time yield of D-mannose reached 27.2 g L-1 h-1 with a maximum conversion ratio of 27%. Therefore, the D-mannose isomerase may be suitable for green production of D-mannose.
Keywords: Characterization; Mannose; Mannose isomerase; Mannose production; Pseudomonas syringae.