Rb-1-hydrolyzing beta-glucosidase from Aspergillus niger KCCM 11239 was studied to develop a bioconversion process for minor ginsenosides. The specific activity of the purified enzyme was 46.5 times greater than that of the crude enzyme. The molecular weight of the native enzyme was estimated to be approximately 123 kDa. The optimal pH of the purified enzyme was pH 4.0, and the enzyme proved highly stable over a pH range of 5.0-10.0. The optimal temperature was 70 degrees C, and the enzyme became unstable at temperatures above 60 degrees C. The enzyme was inhibited by Cu2+, Mg2+, Co2+, and acetic acid (10 mM). In the specificity tests, the enzyme was found to be active against ginsenoside Rb-1, but showed very low levels of activity against Rb-2, Rc, Rd, Re, and Rg(1). The enzyme hydrolyzed the 20-C,beta-(1 -> 6)-glucoside of ginsenoside Rb-1 to generate ginsenoside Rd and Rg(3), and hydrolyzed 3-C,beta-(1 -> 2)-glucoside to generate F-2. The properties of the enzyme indicate that it could be a useful tool in biotransformation applications in the ginseng industry, as well as in the development of novel drug compounds.