The Glycosidase Treatment of Gentianae Scabrae Radix Converts Trifloroside into Deglucosyltrifloroside with an Enhancement of Antioxidative Effects.

ABSTRACT

Herbal medicines were subjected to enzyme reaction by using a commercial glycosidase AMG-300L, and were evaluated for enhancement of their antioxidative activities. The methanolic extract of Gentianae Scabrae Radix (GSR) showed the most dramatic changes after enzyme reaction, as seen in the high-performance liquid chromatography profiles and an increase in the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging effect. Trifloroside (1, TF) was identified as being significantly decreased by enzyme reaction, whereas deglucosyltrifloroside (2, DTF) increased. The optimal reaction time to induce DTF was 24 h at 30°C. The content increased from 1.00 ± 0.29 mg/g of extract (gex) to 2.80 ± 0.85 mg/gex after 24 h of enzyme reaction. DTF showed better antioxidative effect than TF in the DPPH, Trolox equivalent antioxidant capacity, and reactive oxygen species (ROS) in HT22 cell assays. In addition, when HT22 cells were stressed by 5 mM glutamate, 50 µM of DTF significantly inhibited the glutamate-induced lactate dehydrogenase leakage, Ca2+ influx, lipid peroxidation, and intracellular ROS production. These data demonstrated that the enzyme-treated GSR and its increased level of antioxidant DTF could be useful as a starting point in the discovery of functional foods to prevent various oxidative stresses, especially neurodegenerative disorders.