玉米ACE抑制肽的制备工艺及中试生产

Preparation of ACE-inhibitory peptide from corn gluten meal and pilot-scale production

以玉米黄粉为原料，利用Alcalase酶、Protamex酶和Flavourzyme酶酶解制备血管紧张素转换酶（ACE）抑制肽，研究酶解条件、酶解方式对玉米肽ACE抑制活性的影响。同时，在试验的基础上，对100kg玉米黄粉酶解制备ACE抑制肽进行了中试生产。实验结果表明：单酶水解制备ACE抑制肽的最适酶为Protamex酶，在酶解150rain时水解产物的ACE抑制率为（90．98±0．54）％，水解产物的分子质量集中在10405．22—300Da。Flavourzyme酶和Protamex酶的分步水解可进一步提高玉米蛋白的酶解效率和玉米肽对ACE的抑制活性，在先用Flavourzyme酶水解2h再加入Protamex酶继续水解3h后，水解产物的ACE抑制率达到（99．35±3．55）％（IC50=0．764mg／mL），水解产物的分子质量集中在8439．49～185．2Da。采用双酶分步水解方式水解100kg玉米黄粉时，肽粉得率为27％，ACE抑制率为（56．70±0．86）％（1mg／mL）。

In the present study, corn gluten meal （CGM） was hydrolyzed using three proteases （Alcalase,Protamex and Flavourzyme） to produce the ACE-inhibitory peptide. Effect of hydrolysis conditions and methods on ACE- inhibitory activities of corn peptide was investigated. Meanwhile, on the base of small scale experiment, ACE-inhibitory peptide from hydrolyzing 100 kg CGM was prepared. The results showed that Protamex protease was the optimal enzyme for the production of ACE-inhibitory peptide when using one-step hydrolysis, and under the condition of 150 min of hydrolysis, the inhibition rate of ACE was 90.98% ±0.54% , the molecular weight of peptides in the hydroly- sate was mainly distributed from 300 Da to 10 405.22 Da. The hydrolysis efficiency of corn protein and ACE-inhibitory activity of corn peptides were increased by two-step enzymatic hydrolysis of Flavourzyme and Protamex proteases. 1The first-step enzymatic hydrolysis with Flavourzyme proteases was carried out for 2 h, the mixture was subjected to a second-step enzymatic hydrolysis with Protamex proteases for another 3 h, the ACE inhibition activity of hydrolysate reached to 99.35% ± 3.55% （ICs0 =0. 764 mg/mL） , the molecular weight of peptides was mainly distributed from 185.2 Da to 8 439.49 Da. In pilot-scale production, two-step enzymatic hydrolysis was applied to prepare ACE-inhibitory peptide, yield of peptide power was 27% and the inhibition rate of ACE was 56.70% ±0.86% at 1 mg/mL.