Various glucobioses were synthesized enzymatically by two procedures, an incubation of a highly concentrated solution of D-glucose in the presence of glycosidases (batch method reaction), and the other is circulation of highly concentrated D-glucose solution through an immobilized glycosidase column and an activated carbon column connected in series (column method reaction). The composition of disaccharides differed with the difference of reaction methods and with the kind of glycosidases. A similar reaction was also done with glucoamylase and the factors controlling the regioselectivity of the reaction with the differences of enzyme or with the differences of the procedures are discussed.