The present application relates to the field of
metabolic engineering, and provide a method for high-efficiency synthesis of D-allulose by
Escherichia coli using nutritional sweetener-
sucrose, wherein D-allulose is produced by
recombinant Escherichia coli using cheap and traditional nutritional sweetener, and wherein
Escherichia coli JM109(DE3) is used as a
chassis host, wherein a path for producing D-allulose from
sucrose is constructed by introducing exogenous proteins
sucrose-6-
phosphate hydrolase CscA, sucrose permease CscB, fructokinase CscK, D-allulose-6-
phosphate epimerase AlsE and D-allulose-6-
phosphate phosphatase A6PP into wild-type
Escherichia coli to realize co-expression, and wherein the substrate
utilization rate is improved by introducing ptsG-F and knocking out ptsG, fruA, ptsI and ptsH to reprogram the
sugar transmembrane transport pathway.