Lactose has become the main byproduct of many dairy products and ingredients. Current applications of lactose are insufficient to use the recovered lactose from manufacturing operations. Here we exemplified a new process for converting aqueous lactose into a sweeting syrup via one-pot synthesis. The synthesis consisted of two-steps: (1) enzymatic hydrolysis of lactose and (2) catalytic isomerization over MgO/SiO2. The hydrolysis of lactose over β-galactosidase converted 95.77 ± 0.67% of lactose into glucose and galactose. The catalytic isomerization was performed over MgO/SiO2 with different MgO loadings (10-40 wt.%). A battery of tests was conducted to characterize the different catalysts, including surface properties, basicity, and microstructure. The one-pot synthesis, enzymatic hydrolysis and catalytic isomerization over 20%-MgO/SiO2, converted 99.3% of lactose into a sweetening syrup made of glucose (30.48%), galactose (33.51%), fructose (16.92%), D-tagatose (10.54%), and lactulose (3.62%). The outcomes of this research present an opportunity for expanding the utilization of lactose.