<p>The research represents first scientific exploration into pectinase-assisted enzymatic hydrolysis of bee pollen, targeting its structurally resilient exine and intine layers. The disruption of these stable layers is crucial in promoting the nutrient release (amino acids, bioactive compounds, and minerals) and potentially broadening its applicability in numerous food formulations. Response surface methodology ascertained that optimal parameters for effectively disintegrating bee pollen cell walls are an enzyme concentration of 0.26%, a pH of 4.6, a temperature of 48.7℃ and hydrolysis time of 12&#xa0;h with protein dispersibility index, wall-breaking rate, 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, and total phenolic content as the response variables. Furthermore, the artificial neural network model (R² = 0.99) successfully validated the experimental data obtained from response surface methodology, ensuring robust predictive accuracy. The scanning electron micrographs of pectinase optimized bee pollen (PEOP) demonstrated complete disruption of cell wall. Subsequent analysis demonstrated a marked increase in crude lipid content (12.43 ± 0.19%), protein (32.14 ± 0.28%), water holding capacity (1.95 ± 0.02%), emulsifying activity (65.58 ± 1.35%) compared to untreated bee pollen. Significant increase was also observed in essential amino acids (1.5 times), minerals (1.1 times), in vitro digestibility of PEOP with reduced thermal stability. Minimum alterations in functional group and degree of crystallinity confirms the integrity of the PEOP, ensuring its suitability as a functional food supplement. Therefore, the results strongly establish that pectinase hydrolysis is a productive approach to disrupt bee pollen cell wall for maximising the nutrient release, and bioavailability hence, paving the way for the utilization of fragmented nutrient-rich bee pollen in diverse food applications.</p>