Impact of green techniques on intricate cell wall structure of bee pollen to enhance functional characteristics and improve its in vitro digestibility

Bee pollen is a nutrient-rich super food, but its rigid dual-layered structure limits nutrient release and absorption. The outer exine, composed of stress-resistant sporopollenin, and the inner intine, consisting of cellulose and pectin, form a barrier to digestive breakdown. This study investigates the potential of green techniques, specifically supercritical fluid extraction and ultrasonication, to disaggregate pollen cell walls, enhancing its bioavailability and maximizing nutrient utilization. Ultrasonication treated pollen (USTP) and supercritical fluid extraction-treated pollen (STP) demonstrated disruption, as evidenced by scanning electron microscopy imaging. In relation to scanning electron microscopy, techno-functional, antioxidant, and compositional analysis displayed a positive outcome, with crude lipid, protein, antioxidant activity (2,2-diphenyl-1-picrylhydrazyl activity and 2,2'-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid assay) and total phenolic content increased by 34.80%, 32.58%, 10.80%, 11.37%, and 83.94%, respectively. Based on the above properties, USTP for 4 h and STP at 400 bar for 40 min were identified as the optimal conditions for disintegration. Furthermore, optimized samples analyzed for amino acid and mineral release revealed a notable increase in composition of essential amino acid and minerals (Ca, Cu, Fe, etc.) by ∼1.5 and 1.2 times, respectively. Along with significant changes in composition, fractured pollen exhibited 1.4 folds increase in protein digestibility with minor differences in thermal stability, and crystallinity as established by differential scanning calorimetry, and X-ray diffraction analysis. The study confirms that nutrient release and absorption remain restricted without pre-treatment, highlighting the necessity of specific treatment to disintegrate bee pollen before its use as a functional food ingredient. PRACTICAL APPLICATION: Bee pollen is a rich source of all the essential nutrients required by the humans and recognised as a complete food. However, its tough cellular structure restricts its utilisation in numerous food applications. Therefore, to disintegrate bee pollen and release its nutrients, ultrasonication and super critical fluid extraction processes were employed to improve its utilization for human purposes. Both the treatment techniques, enhanced bee pollen's bioavailability and functional properties, making it more suitable for use in nutraceuticals and functional foods.These treatments proved to increase the antioxidant capacity, digestibility, and create high-value ingredient for supplements, beverages, and fortified foods.