Wang_2026_3.Biotech_16_219

A robust and reusable Rhizomucor miehei lipase (RML) biocatalyst was prepared by integrating bio-imprinting with resin immobilization. RML was co-imprinted with lauric acid and polyethylene glycol (PEG600) followed by immobilization on an amine-functionalized epoxy resin (ES-103B), resulting in AE@RML-LA+PEG600. After immobilization process optimization, the AE@RML-LA + PEG 600 was verified by CD, SEM, FTIR, and XRD analysis. AE@RML-LA+PEG600 exhibited improved thermal, pH, and organic solvent stability compared with free RML. Under optimized conditions (10 h, 62 degreesC, palmitic-acid-enriched palm stearin/oleic acid = 1:10 mol/mol, 10 wt% enzyme), AE@RML-LA+PEG600 achieved a maximum OPO content of 45.80%, and the proportion of palmitic acid at the sn-2 position relative to total palmitic acid ((sn-2, P)/P) was 54.59%, which was higher than that of the unimprinted enzyme (35.52% OPO). After five cycles, AE@RML-LA+PEG600 maintained 69.52% of its original catalytic activity, indicating its excellent operational stability. This work provides a promising imprinting-immobilization strategy for preparing durable lipases for industrial OPO production. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-026-04860-6.