Ginsenoside compound K was produced from ginseng root extract using a thermostable recombinant β-glycosidase from Sulfolobus solfataricus. The pH and temperature for maximum production were 5.5 and 90 °C. The half-lives of the enzyme were 66, 30, and 1.7 h at 80, 85, and 90 °C respectively. The ginsenoside-hydrolyzing activity (Rd>Rb₁>Rc>Rb₂) and compound K-producing activity (Rd>Rc>Rb₁>Rb₂) of the β-glycosidase were also determined. At pH 5.5 at 85 °C, 1.63 mg/ml of compound K was produced within 12 h by 40 U/ml of enzyme from 1.9 mg/ml of ginsenoside Rb₁, 0.52 mg/ml of ginsenoside Rb₂, 0.92 mg/ml of ginsenoside Rc, and 0.23 mg/ml of ginsenoside Rd in a 10% (w/v) ginseng root extract via two transformation pathways, Rb₁ or Rb₂ → Rd → F₂ → compound K, and Rc → compound Mc → compound K.

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