Fig. 4: Investigation of increased enzyme stability and CE in the enzyme@IP system.

a Schematic of CALB in various environments: I water-enriched enzyme@IP, II water-free enzyme@IP, and III CALB adsorbed onto microspheres. b Conversion of enzymatic epoxidation over time in different flow systems. The reaction conditions are the same as those in Fig. 3c. c CE of CALB in different continuous-flow systems. d Fluorescence spectra of CALB in phosphate buffered saline (PBS) and ethyl acetate (EA), both containing 0.5 mol L⁻¹ H₂O₂, before and after standing for 24 h. e Circular dichroism (CD) spectra of CALB after 24 h treatment in different PBS/EA mixtures, including PBS, a mixture of 33% (v/v) PBS in EA, and pure EA (all containing 0.5 mol L⁻¹ H₂O₂). f Secondary structure changes of CALB after 24 h treatment in various PBS/EA mixtures, including α-helices, β-sheets, β-turns, and random coils. g Raman spectral intensity profiles showing the H₂O₂ concentration distributions from the exterior to interior. h Schematic illustration showing H₂O₂ diffusion through the hydrophobic pores of the “interphase”. i CE comparison of CALB with different “interphase” hydrophobicity. j Comparation of residual enzyme activity between hydrophilic and hydrophobic “interphase” after 12 h incubation in H₂O₂ solutions with varying concentrations from 0.5 to 1, 2 and 4 mol L⁻¹. k Comparison of kinetic parameters (K_m, V_max and k_cat) for enzymes incorporated in “interphase” with varying levels of hydrophobicity.