Tailoring ω-3 fatty acid enrichment through genipin, glutaraldehyde, and glyoxyl linked immobilization of Rhizomucor miehei lipase on MWCNTs

, while MWCNT/Gen@RML exhibited superior reusability, retaining 75% of its initial activity after five cycles. During fish oil hydrolysis, MWCNT/Gen@RML provided the fastest midterm accumulation (92.85% total EPA + DHA at 36 h), whereas Glu-RML delivered the highest endpoint yield (82.33% at 48 h). The immobilized systems also enabled selective enhancement of either DHA or EPA, depending on coupling agent type and enzyme loading. Overall, the use of functionalized MWCNTs as supports, combined with mild natural and synthetic coupling agents produced stable high-performing lipase biocatalysts, and offered a sustainable route for the concentration of ω-3 fatty acids under mild, environmentally friendly conditions.