This invention provides a method for preparing
immobilized lipase using composite-modified
protein amyloid protofibrils. The method first prepares β-lactoglobulin
amyloid protofibrils by controlling pH and hydrothermal conditions, then polymerizes them into COF protofibrils via TFB and DB condensation. An immobilized carrier is obtained by composite modification with sulfopropyl
betaine and pentamalloylmannose to adsorb and immobilize the
lipase. In this invention, sulfopropyl
betaine anchors to the carrier surface through electrostatic,
hydrogen bonding, and hydrophobic interactions, forming a hydration film to prevent pore blockage and enhance the carrier's resilience. Pentamalloylmannose permeates the pores to form a stable interpenetrating structure, giving the carrier a high specific surface area and stability. Furthermore, it stabilizes the loaded
lipase through multiple interactions, preventing conformational damage and detachment of the
enzyme, while simultaneously creating a stable microenvironment for the
lipase, inhibiting oxidation and denaturation inactivation. The
immobilized lipase prepared by this invention exhibits high catalytic activity, excellent substrate
mass transfer efficiency, and good
reusability, and can efficiently catalyze
oleic acid esterification reactions.