Combinatorial Enzymatic Catalysis for Bioproduction of Ginsenoside Compound K

Ginsenoside compound K (CK) is an emerging functional food or pharmaceutical product. To date, there are still challenges to exploring effective catalytic enzymes for enzyme-catalyzed manufacturing processes and establishing enzyme-catalyzed processes. Herein, we identified three ginsenoside hydrolases BG07 (glucoamylase), BG19 (β-glucosidase), and BG23 (β-glucosidase) from Aspergillus tubingensis JE0609 by transcriptome analysis and peptide mass fingerprinting. Among them, BG23 was expressed in Komagataella phaffii with a high volumetric activity of 235.73 U mL^-1 (pNPG). Enzymatic property studies have shown that BG23 is an acidic (pH adaptation range of 4.5-7.0) and mesophilic (thermostable < 50 °C) enzyme. Moreover, a one-pot combinatorial enzyme-catalyzed strategy based on BG23 and BGA35 (β-galactosidase from Aspergillus oryzae) was established, with a high CK yield of 396.7 mg L^-1 h^-1. This study explored the ginsenoside hydrolases derived from A. tubingensis at the molecular level and provided a reference for the efficient production of CK.