Hydrogen-bonded organic framework-based immobilization of β-glucosidase for sustainable and high-yield resveratrol...

β-Glucosidase (BGL) is a key biocatalyst widely used for glycosidic bond hydrolysis in food, biomass, and pharmaceutical applications. Industrial processes for certain high-value glycoside transformations often involve harsh conditions, including acidic and elevated-temperature, to enhance substrate solubility and reaction rates. However, under these conditions, free and conventionally immobilized β-glucosidase often suffer rapid loss of catalytic activity and limited reusability. Herein, we report, to our knowledge, the first application of a hydrogen-bonded organic framework (HOF) for β-glucosidase immobilization to overcome these challenges. The β-glucosidase was aminated with carbodiimide as the carrier and immobilized on HOF101. Under the amination conditions of an initial enzyme concentration of 2 mg/mL and pH 4.5, the immobilization efficiency reached 85.3 % and the activity retention rate reached 93 %. After immobilization, the optimal pH remained unchanged, and the optimal temperature dropped from 60 °C to 50 °C. Notably, under low-temperature (30 °C) conditions, the enzyme activity increased from 36.58 % to 90 %, enhancing its application activity at low temperatures. The immobilized enzyme retained 94.1 % of its initial activity after eight weeks of storage and 59.6 % after ten catalytic cycles under acidic, elevated-temperature conditions. Under these challenging conditions, the immobilized enzyme achieved 97.7 % conversion of polydatin to resveratrol within 1.5 h. This approach not only enhances β-glucosidase performance in demanding reaction environments but also provides a versatile, recyclable, and sustainable platform for immobilizing other enzymes requiring improved stability in industrial biocatalysis.