Fig. 3: Functional, biochemical characterization, and structural investigation of PnGH1. | Nature Communications

Fig. 3: Functional, biochemical characterization, and structural investigation of PnGH1.

From: Discovery of plant chemical defence mediated by a two-component system involving β-glucosidase in Panax species

Fig. 3

a SDS–PAGE analysis, HPLC chromatogram, and kinetic study of the PnGH1-conducted hydrolysis reaction of Rb3 to form Fd. Kinetic assays were performed in independent duplicates. b Subcellular localization investigation of PnGH1. The experiment was repeated independently three times with similar results. c The overall structure of PnGH1 with the labeled (β/α)8 barrel. d Surface view of the binary complex structure of PnGH1 docking with Rg3 showing the widely open entrance of the substrate binding pocket. The distance between E425 and E232 (5.0 Å) was shown as a purple dash. e Docking and simulation results of key residues in ginsenosides substrates binding pocket of PnGH1. Rg3 and key residues were shown as yellow and cyan sticks, respectively. Hydrogen bonds and hydrophobic interactions were shown as purple and green dashes, respectively. f The retaining reaction mechanism of the hydrolysis of PPD-type ginsenosides catalyzed by PnGH1 through the double displacement mechanism. g The relative activity of different mutants of PnGH1. Data are presented as the mean ± SD of triplicate independent experiments. Source data are provided as a Source Data file.

Back to article page