Dual Lipase Immobilization on Kaolin‐Based Support for High Catalytic Efficiency and Long‐Term Stability

Abstract The co‐immobilization process of lipases in kaolin enhances industrial processes, making them more efficient, sustainable, economical, and innovative. This study aims to optimize the co‐immobilization of CALB and Eversa Transform 2.0 lipases in functionalized kaolin, envisioning the subsequent computational analysis of enzyme‐support interactions. The statistical tool applied for process optimization was the Taguchi L9 method, which enabled the identification of the ideal conditions for enzyme immobilization (time: 24 h, pH 9, buffer concentration: 5 mM, and enzyme load: 6 mg g −1 of support) in modified kaolin. Subsequently, the produced biocatalyst was characterized by X‐ray diffraction (XRD), Fourier transform infrared (FTIR) spectroscopy, thermogravimetry (TGA), and scanning electron microscopy (SEM). The KCBG/Eversa‐CALB biocatalyst showed superior thermal stability, with a half‐life 2 to 3 times longer than soluble enzymes between 50 °C and 80 °C, in addition to resistance to pH and solvent variations. After 180 days, it maintained more than 85% of the initial activity. In addition, it demonstrated high enantio, regio‐, and stereoselectivity for secondary alcohols and retained 50% of its activity after five cycles. In summary, the co‐immobilization of CALB and Eversa in kaolin improved the operational stability of the enzymes, increasing the efficiency and sustainability of industrial processes.