Abstract:[Objective] This study aimed to determine the optimal extraction process for antihypertensive peptide from Agrocybe cylindracea to achieve higher yield and enhanced activity.[Methods] With the polypeptide yield and ACE inhibition rate as the evaluation index,the effects of enzyme type,substrate concentration,enzyme concentration,pH value,reaction time,reaction temperature and other single factors on the antihypertensive peptide and its activity were investigated,and the optimal extraction process of antihypertensive peptide was optimized by response surface.[Results]] The optimal extraction conditions were as follows:substrate concentration of 5.4%,enzymolysis duration of 8 h,reaction temperature was 23.3 ℃,pH 8.0,and neutral protease dosage of 2 g.Under the control of these conditions,the antihypertensive peptide yield reached 92.12%,with an ACE inhibition rate of 93.75%.The active peptide fraction,primarily composed of peptides with molecular weight <3 kDa,accounted for 36.69% of the total peptide and demonstrated an ACE inhibition rate of 95.36%.[Conclusion] The extraction process for antihypertensive peptides from A.cylindracea was successfully optimized,and the molecular weight distribution of the active peptide components was determined.