双酶水解螺旋藻藻胆蛋白制备ACE抑制肽的工艺优化

Optimization of Preparation Process of Spirulina Acetylcholine ACE Inhibitory Peptide by Double Enzyme Hydrolysis

以螺旋藻为原料,采用反复冻融和超声波解冻结合法提取藻胆蛋白,利用等电点分离藻胆蛋白,采用SDS-PAGE电泳确定分离蛋白的种类。采用胃蛋白酶和胰蛋白酶双酶先后水解藻胆蛋白。通过单因素实验和正交实验优化胃蛋白酶和胰蛋白酶水解藻胆蛋白制备血管紧张素转换酶(ACE)抑制肽的工艺,研究表明:胃蛋白酶水解藻胆蛋白最佳酶解工艺条件是水解温度为37℃,底物质量浓度为6%(w/v),酶与底物比为5220 U/g,此时ACE抑制率为82.07%。胰蛋白酶水解藻胆蛋白最佳酶解条件是温度为42℃,底物质量浓度为6%(w/v),酶与底物比为5220 U/g,此时ACE抑制率为80.35%。利用超滤离心管获得分子量小于3 kDa的藻胆蛋白ACE抑制率94.30%。

Phycobiliprotein was extracted from spirulina by repeated freeze-thaw and ultrasonic de-freezing,and separated by isoelectric point. The type of protein isolated was determined by SDS-PAGE electrophoresis. The phycobiliprotein was hydrolyzed successively by pepsin and trypsin. The process of preparing angiotensin-converting enzyme(ACE)inhibitory peptide by pepsin and trypsin hydrolysis of phycobiliprotein was optimized by single factor experiment and orthogonal experiment. The results showed that the optimal enzymatic hydrolysis conditions of pepsin hydrolyzed phycobiliprotein were hydrolysis temperature 37 ° C,the substrate mass concentration 6%(w/v),and the enzyme to substrate ratio 5220 U/g.Under the conditions,the ACE inhibition rate was 82.07%. The optimal enzymatic hydrolysis conditions for trypsin hydrolysis of phycobiliprotein were temperature 42 ℃,substrate mass concentration 6%(w/v),enzyme to substrate ratio 5220 U/g. Under the conditions,ACE inhibition rate of 80.35%. The ACE inhibition rate of phycobiliprotein with a molecular weight of less than 3 kDa was 94.30% using an ultrafiltration centrifuge tube.