Deep eutectic solvent-based extraction of honeybee venom proteins: A green strategy for enhanced recovery and...

Honeybee venom (BV) is a complex biological matrix rich in bioactive proteins and peptides with significant therapeutic potential. Efficient extraction and preservation of these components are essential for improving their bioavailability and functional applications. In this study, protein recovery from bee venom was comparatively evaluated using deep eutectic solvent (DES)-based systems, with an aqueous NaCl solution (0.9%, w/v) used as the baseline. Five choline chloride-based DES formulations were evaluated, among which urea-based systems demonstrated the most promising performance, including enhanced melittin extraction, high pharmacological activity, and particle sizes up to 500 nm with heterogeneous distribution. In addition to peptide enrichment, DES extracts exhibited strong inhibitory activity against protease and α-amylase enzymes, suggesting potential utility in inflammation- and diabetes-related disorders. Hemocompatibility assays further indicated that DES-derived extracts could offer a safer delivery system for intravenous administration compared with NaCl-based system. Collectively, these findings highlight DES systems as selective and efficient media for recovering functional BV proteins while maintaining their bioactivity and biocompatibility. Overall, this work provides evidence that DES-based extraction represents a versatile and sustainable approach for the green recovery of bioactive bee venom components, providing a foundation for future development in pharmaceutical and nutraceutical applications.