Compounds of isolated from roots extract of Pueraria thunbergiana were tested their inhibitory effects on α-glucosidase and α-amylase. Inhibitory activity of methylene chloride (MC) fraction and ethyl acetate (EA) fraction against α-glucosidase showed more than 60% at a concentration of 500μg/ml. Among the nine compounds tested on α-glucosidase, biochanin A, (-)-tuberosin and calycosin from MC fraction and daidzein from EA fraction were stronger inhibitors than acarbose (IC50=530μg/ml), and their IC50 were 9, 144, 328 and 20μg/ml, respectively. Biochanin A and (-)-tuberosin also inhibited α-amylase activity as like as acarbose IC50=20.5μg/ml), and their IC50 were 22 and 348μg/ml, respectively. Although daidzein was already known α-glucosidase inhibitory effects, it was newly evaluated that biochanin A and (-)-tuberosin inhibited α-glucosidase as well as α-amylase, and that calycosin did α-glucosidase.