Characterization Of Two Ginsenoside-glycosidases

This paper mainly concerns the purification and reaction characters of two ginsenoside-glycosidases from microorganism Absidia sp.G4r、Absidia sp.G8r, and ascertains molecular weight, speciality of purified enzyme and kinetics of enzyme reaction.The two glycosidases were pured using DEAE-cellulose column. The pure enzyme purity and molecular weights of the there glycosidases were determined using SDS-polyacrylamide gel electrophoresis to obtain one spot and two ginsenoside-glyosidases were71kDa、72kDa.The optimum pH and temperature for both enzymes catalysis were5.0and30℃respectively. K+、Na+、Mg2+、Zn2+、Ca2+ions had no effect on the enzyme activity,while Cu2+inhibited the enzyme activity.both enzymes can hydrolyze terminal α-Gal、β-Glc on the linkages.The ginsenoside-glyosidase (GluGF-4) from Absidia sp.G4r and the ginsenoside-glyosid-ase (GluGF-8) from Absidia sp.G8r were purified by DEAE-CelluloseDE52and electrophore-sis. The two ginsenoside-glyosidases from Absidia sp.G8r and G4r are the same.The enzyme from Absidia sp.G8r hydrolyzed Rb1to C-K, first, hydrolyzed Rb1to Rd, about8h,hydrolyzed Rd to F2,about18h, hydrolyzed F2to C-K.The Vmax of the enzyme are1.524L·h·mmol-1、0.904L·h·mmol-1、0.447L·h·mmol-1,the Km of the enzyme are9.253mmol·L-1、5.488mmol·L-1、5.247mmol·L-1.