The 1-deoxynojirimycin (DNJ) biosynthetic gene cluster of Bacillus amyloliquefaciens 140N isolated from traditional Korean fermented food was isolated by PCR screening. It showed 78.9% inhibitory activity against α-glucosidase and produced 0.8 g/L of DNJ in an optimized medium containing 2% soluble starch, 1% tryptone, 0.05% KH(2)PO(4), and 0.05% (NH(2))(4)SO(4). Soybean paste fermented with B. amyloliquefaciens 140N produced DNJ with 84.4% inhibitory activity.