Rare ginsenoside CK is in high demand due to its significant physiological activity, but its low natural abundance limits applications. While metal-organic framework (MOF)-based immobilized enzyme technology enables the conversion of ginsenoside Rb1 to CK, it remains largely confined to laboratory scale. In this study, a zirconium-based MOF (UiO-66-Ser) was greenly synthesized in a choline chloride-ethylene glycol deep eutectic solvent (DES) using L-serine as a modulator for the co-immobilization of snailase and β-glucosidase. This co-immobilized system achieved a CK yield of 88.89 % at the laboratory scale. To evaluate scalability, systematic investigations were conducted on the effects of scaling up the synthesis system (20–60 mL), cross-linking system (20–200 mL), and conversion system (1–100 mL). The results indicate that after large-scale production, immobilized enzymes consistently exhibit superior thermal stability, pH stability, and organic solvent stability compared to free enzymes. Even at the maximum scale (60 mL synthesis, 200 mL cross-linking), the CK yield remained around 65 % (65 ± 0.38 %). The conversion system achieved peak efficiency at 10 mL, with a CK yield reaching 90.41 %, and the immobilized enzyme showed a more gradual decline in yield than free enzymes during scale-up. This study confirms the excellent scalability of the DES-synthesized Zr-MOF-based dual-enzyme immobilization system, providing a theoretical foundation and practical support for the industrial application of efficient biotransformation of natural products.
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