The purpose of the present invention is to provide a technology whereby, in a method for manufacturing a sweetener composition containing glucose, fructose and allulose, said method comprising treating glucose with glucose isomerase and allulose epimerase, the content of allulose in the sweetener composition is increased. A sweetener composition containing glucose, fructose and allulose and having a high allulose content can be continuously manufactured at a high efficiency by immobilizing glucose isomerase and allulose epimerase, packing the same into a column so as to give an activity ratio of the immobilized glucose isomerase to the immobilized allulose epimerase of 1.49:1-5.61:1, and then passing a glucose solution through the column.
