大豆蛋白体外酶解物中血管紧张素转化酶抑制剂活性肽研究

目的:体外模仿部分胃肠道消化酶解过程,考察大豆蛋白酶解消化能否产生血管紧张素转化酶抑制剂(ACEI)活性肽及其活性状况,以揭示大豆蛋白体内消化酶解与ACEI的活性关系。方法:模拟人体胃肠道消化过程,以胃蛋白酶结合胰蛋白酶,相继酶解大豆分离蛋白(SPI),经色谱分离,动态检测不同阶段ACEI肽片段及其活性大小。结果:胃蛋白酶酶解过程前20min内,酶解液ACEI活性达到最高点,随后在胰蛋白酶酶解阶段其抑制活性下降。180min后的酶解产物,其半抑制活性浓度IC50值为0.28±0.06 mg/ml。同时,未经酶解的SPI液在0.73mg/ml时无ACEI活性。SPI酶解液经各种色谱分离后的组分,其IC50值从0.13±0.03到0.93±0.08 mg/ml。低分子量和伴有疏水性基团的肽类最具ACE抑制活性。结论:体外模仿胃肠消化过程使用胃蛋白酶和胰蛋白酶酶解SPI可产生不同ACEI活性的肽片段,说明人体正常摄食消化大豆蛋白可产生血管紧张素转化酶抑制剂活性肽。

STUDY ON ANGIOTENSIN CONVERTING ENZYME INHIBITORY PEPTIDES FROM DIGESTION OF SOY PROTEIN IN VITRO

Objective: To investigate if angiotensin converting enzyme inhilitory(ACEI) peptides would be produced from SPI digested by a batch digestion system using enzymes similar to digestive enzymes in humans.Method: Simulate the conditions of human gastrointestinal digestion in a model digestion system in vitro and produce soy peptides from SPI digested using pepsin and pancreatin.In addition to monitoring ACEI activity in the total soy protein digest,the possibility of generating soy peptide fractions with more potent activity than the unfractionated digest was investigated by measuring activity of fractions obtained after ultrafitration,anion exchange,and RP-HPLC.Results: The generation of ACEI activity in SPI was determined after sequential digestion with pepsin and pancreatin.The inhibitory activity was highest within the first 20 min at pepsin digestion and decreased upon subsequent digestion with pancreatin.An IC50 value of 0.28±0.06 mg/ml was determined after 180 min of digestion,while no ACEI activity was measured for the undigested SPI at 0.73 mg/ml.Chromatographic fractionation of the SPI digest resulted in IC50 values of active fractions ranging from 0.13±0.03 to 0.93±0.08 mg/ml.Conclusion: Many different peptides with ACEI activities were produced after pepsin-pancreatin digestion of SPI in vitro and lead to the speculation that physiological gastrointestinal digestion could also yield ACE inhibitory peptides from SPI.