In this study, two acidic ionic liquids were innovatively selected to replace traditional acetic acid as catalysts and templates for the preparation of covalent organic frameworks (COFs) materials in aqueous phase at room temperature. At the same time, the synthesized COFs carrier achieved in-situ encapsulation and immobilization of Candida rugosa lipase (CRL) in aqueous phase at room temperature. The optimum immobilized lipases COF-COOH-M and COF-SO3H-M were obtained by adjusting the reaction conditions. Their enzyme loadings were 210.17 mg/g and 258.47 mg/g, respectively, and the specific activity retention compared to free lipase was 67.11% and 78.42%, respectively. The carrier and immobilized lipase were characterized by FT-IR, XRD, BET and TG. The catalytic performance and operational stability of the immobilized lipase were investigated. The results showed that both immobilized lipases showed better stability than free lipases, and COF-SO3H-M had the best performance. The relative activity retention of COF-SO3H-M after incubation at 60 °C for 60 minutes was 4.35 times that of free lipase. After 28 days of storage at 4 °C, the relative activity of COF-SO3H-M remained 75.10%, while that of free lipase was only 24.61%. The COF-SO3H-M was applied as biocatalyst for isoamyl acetate preparation at 50 °C for 7 hours, and the yield of isoamyl acetate was as high as 86.94%, which was 22.12% higher than that of free lipase.
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