According to the D-
psicose 3-epimerase
mutant and the application thereof, the D-
psicose 3-epimerase
mutant is obtained through
protein C-terminal sequence modification,
software analysis and
sequence alignment of different sources of D-
psicose 3-epimerase, the half-life period of the
mutant enzyme at 60 DEG C, 70 DEG C and 80 DEG C is prolonged, and the activity of the
mutant enzyme is improved. The substrate affinity and conversion efficiency of the
enzyme to D-
fructose as a substrate for synthesizing D-psicose are improved, and the Tm value of the mutant is 73.23 DEG C and is improved by 11.98 DEG C compared with that of a
wild type; the half-life period at the temperature of 60 DEG C is 255.35 min and is 17.60 times that of a
wild type, the problem that the half-life period of the
wild type is too short at the high temperature is solved, the relative action time of the
mutant enzyme is long, the yield of the product D-psicose is remarkably increased by applying the
genetically engineered bacterium containing the modified
enzyme for
biotransformation, the product yield is increased by 4.3%, the equilibrium conversion rate reaches 33%, and the yield of the product is greatly increased. And a green, environment-friendly, sanitary and safe biological preparation method is provided for the preparation process of the D-psicose.