酶解法制备黑木耳ACE抑制肽的工艺优化

Optimization of ACE Inhibitory Peptide Preparation from Auricularia auricula by Complex Enzymatic Hydrolysis

摘要:

以黑木耳为原料，利用复合酶解法制备血管紧张素转化酶(Angiotensin-Converting Enzyme，ACE)抑制肽。以ACE抑制率和蛋白水解度为指标，通过单因素试验确定复合酶的组合方式及加酶量，利用正交试验对酶解工艺进行优化，优化酶解温度和酶解时间等工艺参数。结果表明，黑木耳的最佳酶解工艺为：料液比1∶50(g/mL)、加酶量200 U/g、复合蛋白酶(中性蛋白酶∶胰蛋白酶)比例1∶2(U/U)、酶解温度55℃、酶解时间4 h、酶解pH值7.5。在此酶解工艺条件下黑木耳蛋白水解度达61.22%，ACE抑制率达65.17%，酶解液中多肽分子量主要集中在131~4 983 Da，占比达92.19%。通过对黑木耳酶解工艺的优化，提高了黑木耳蛋白的水解度及ACE抑制率，为黑木耳制备ACE抑制肽的过程控制及规模应用提供理论指导。

Abstract:

Angiotensin-converting enzyme(ACE) inhibitory peptide was prepared from Auricularia auricula using complex enzymatic hydrolysis. ACE inhibition rate and degree of hydrolysis were used as key evaluation indices to optimize the preparation process through single-factor experiments and orthogonal testing. The optimal enzyme combination, enzyme dosage, hydrolysis temperature, hydrolysis time, and other process parameters for the complex enzyme system were determined. The results indicated that the optimal enzymatic hydrolysis conditions for Auricularia auricula were as follows: a solid-liquid ratio of 1∶50 (g/mL), enzyme dosage of 200 U/g, hydrolysis temperature of 55 ℃, hydrolysis time of 4 hours, pH 7.5, and a neutral protease to trypsin ratio of 1∶2(U/U). Under these conditions, the degree of hydrolysis reached 61.22%, and the ACE inhibition rate was 65.17%. Additionally, 92.19% of the peptides had molecular weights ranging between 131 Da and 4,983 Da. These optimal conditions effectively enhanced protein hydrolysis and increased the ACE inhibition rate in the enzymatic hydrolysate of Auricularia auricula, providing theoretical guidance for process control and the large-scale application of enzymatic hydrolysis in the preparation of ACE inhibitory peptides from Auricularia auricula.