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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 en