This study utilizes deep eutectic solvent (DES)-assisted enhancement of cellulose-based immobilized probiotics for efficient biotransformation of isofraxidin from Acanthopanax senticosus. Among seven probiotic strains tested, Lactiplantibacillus plantarum CICC 20767 exhibited the best catalytic activity. We explored the effects of 12 different DESs with varying hydrogen bond donors on the conversion capacity of the immobilized probiotics, with the highest efficiency observed using ChCl/EG (4.0 wt %). The optimized process, with a solid-to-liquid ratio of 1:5 (g/mL), a temperature of 35.6 °C, a reaction time of 4 d, and a pH of 6.9, resulted in a 5.53-folds increase in isofraxidin yield, reaching 0.4034 mg/g, compared to the untreated sample (0.0729 mg/g). The immobilized probiotics retained excellent catalytic activity after 12 cycles of use, demonstrating their stability and potential for large-scale, green production of isofraxidin. This study presents a valuable method for industrial isofraxidin production and highlights the broad potential of this environmentally friendly bioconversion process in the pharmaceutical industry.
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