Hyperactivation Behavior of Site-Specifically Immobilized Fusion Protein of Lipase with α-Synuclein and Silica-Binding Peptide

-nitrophenyl palmitate (pNPP) assay at 30 °C in 50 mM HEPES buffer (pH 8.0), and relative activity was expressed as the ratio to the wild-type BTL2. It was found that free SiBP-BTL2-αS showed a 1.8-fold higher activity than BTL2 due to the chaperone effect of αS, and immobilization on MSNs brought out a further 1.4-fold increase in activity at an enzyme loading of 194 mg/g. Thus, SiBP-BTL2-αS@MSNs presented 3.3-fold higher activity than BTL2. Moreover, SiBP-BTL2-αS@MSNs exhibited significantly improved thermostability and broad pH tolerance over the free counterpart and BTL2. In repeated uses, SiBP-BTL2-αS@MSNs retained 82.1% of its initial activity after seven consecutive reaction cycles. In the synthesis of vitamin E succinate, SiBP-BTL2-αS@MSNs showed 32% and 78% higher yields over SiBP-BTL2-αS and BTL2, respectively, verifying the superiority of SiBP-BTL2-αS@MSNs in enzymatic catalysis. This work not only offers a highly efficient, robust, and recyclable enzyme preparation, but also provides a promising way to design immobilized lipase with hyperactivation behavior.