Abstract:To explore the synergistic effects of enzymatic hydrolysis,as well as ultrasonic and alkali treatment on the wall disruption and extraction of active ingredients in pine pollen,scanning electron microscopy （SEM）was used to observe the microstructure of pine pollen,and high-performance liquid chromatography was used to analyze the composition and content of phenols. The results showed that the total phenol content,total flavonoid content,and ABTS+ free radical scavenging ability of the soluble part of pine pollen turned to be the highest after the treatment with 12.5% alkaline protease for 60 min,with the specific values reaching （5.97±0.09） mg GAE/g DW pine pollen,（1.65±0.04） mg RE/g DW pine pollen,（95.12±1.84） μmol TE/g DW pine pollen,respectively. Total phenols,total flavonoids,ABTS+ free radical scavenging ability,and iron ion reduction ability increased by 107%,113%,73%,and 81%,respectively compared with untreated pollen after enzymatic hydrolysis combined with ultrasonic and 0.3 mol/L NaOH treatment. The results of SEM revealed that the wall disruption of pine pollen was further intensified after enzymatic hydrolysis and alkali treatment,and the substances in pollen were spilled outward from the clefts,which was consistent with the trends of physicochemical indexes. The optimal process conditions were 0.3 mol/L NaOH ultrasonic and alkali treatment for 1 h,pH of 7,and enzymatic hydrolysis with 12.5% alkaline protease at 50 ℃ for 60 min. Ultrasonic and alkali treatment could assist enzymatic hydrolysis to break the wall of pine pollen effectively,and nutrients such as small peptides,polyphenols,and flavonoids were further dissolved,which promoted the high-value utilization of pine pollen.