Encapsulation of Immobilized β-Glucosidase with Calcium Metal-Organic Frameworks for Enhanced Stability in Hydrolysis of Cellobiose.

β-Glucosidase (β-G) holds promising applications in various fields, such as biomass energy, food, pharmaceuticals, and environmental protection, yet its industrial application is still limited by issues of stability and recycling. Herein, we first immobilized β-G onto the surface of magnetic chitosan nanoparticles (MCS/β-G) through adsorption methods. Subsequently, utilizing the metal-organic framework (MOF), CaBDC, which possesses good stability under acidic conditions, we encapsulated MCS/β-G. The resulting biocatalyst (MCS/β-G@CaBDC) exhibited excellent activity and recyclability. MCS/β-G@CaBDC can convert 91.5% of cellobiose to glucose in 60 min and maintained 81.9% activity after 10 cycles. The apparent K _m value of MCS/β-G@CaBDC was 0.148 mM, lower than free β-G (0.166 mM) and MCS/β-G (0.173 mM). The CaBDC layer increased the mass transfer resistance of the reaction but also triggered structural rearrangement of β-G during the encapsulation process. This resulted in the β-sheet content rising to 68.4%, which, in turn, contributed to enhancing the rigidity of β-G. Moreover, the saturated magnetic strength of this biocatalyst could reach 37.3 emu/g, facilitating its magnetic recovery. The biocatalyst prepared in this study exhibits promising application prospects, and the immobilization method can provide valuable insights into the field of enzyme immobilization.