摘要
采用胃-胰蛋白酶水解蜂王浆主蛋白(MRJPs),对底物浓度、pH值、酶作用时间和蛋白酶浓度对水解效果的影响进行单因素分析,然后通过正交试验对胃-胰蛋白酶共同作用的酶解工艺进行优化;最后采用超滤分离技术对王浆主蛋白水解产物(H-MRJPs)进行分离,以制备血管紧张素转化酶(ACE)抑制肽。结果表明:优化的酶解工艺为37℃恒温、质量比为1%的胃蛋白酶(pH=2.0)和胰蛋白酶(pH=7.5)各水解2h;在最佳工艺条件下,MRJPs水解度为28.7%,总氮回收率为35.5%;通过SDS-PAGE电泳未检测到MRJPs水解产物(H-MRJPs)含有明显的蛋白条带;采用超滤法对H-MRJPs分离得到分子质量范围为<1、1～5和>5ku的3类血管紧张素转化酶(ACE)抑制肽,其ACE半抑制质量浓度(IC50)分别为0.33、0.61和1.09mg.mL-1,即以分子质量<1ku的产物活性最强。该结果为利用王浆主蛋白开发降压功能食品提供了科学依据。
Major royal jelly proteins （MRJPs） were hydrolyzed with pepsin and trypsin, and the effects of substrate concentration, pH value, enzyme response time and proteinase concentration on the hydrolysis efficiency were analyzed, and the orthogonal test analysis of combined action of pepsin and trypsin were done. The enzymolysis products of MRJPs （H-MRJPs） were separated to prepare the angiotensin I-converting enzyme （ACE） inhibitory peptides by ultrafiltration technique. The results showed that the optimized technical parameters were as follows： enzymatic hydrolysis of MRJPs with 1% pepsin at pH 2.0 for 2 h, then 1% trypsin at pH 7.5 for 2 h at 37℃. The degree of hydrolysis and total nitrogen recovery of MRJPs under the optimal condition were 28.7 % and 35.5 % , respectively. No obvious protein bands were shown for enzymolysis products of MRJPs （H-MRJPs） by sodium dodecyl sulfate-polyaerylamide gel electrophoresis （SDS-PAGE）. Three kinds of ACE inhibitory peptides, ranging from 〈1, 1-5 and 〉 5 ku, were separated by ultrafiltration separation of H-MRJPs. The 50% inhibitory concentration （IC50） values of the three kinds of ACE inhibitory peptides were 0.33, 0.61, and 1.09 mg. mL-1, respectively, which exhibited that the peptides with molecular mass 〈 1 ku possessed most high ACE inhibitory activity. The results above provide a scientific basis for developing functional food of antihypertension.
出处
《浙江大学学报（农业与生命科学版）》
CAS
CSCD
北大核心
2012年第4期511-518,共8页
Journal of Zhejiang University:Agriculture and Life Sciences
基金
国家高技术研究发展计划"863"资助项目(2007AA10Z324)
浙江省公益技术应用研究计划资助项目(2011C22029)
杭州市科技发展计划资助项目(20110232B52)
关键词
王浆主蛋白
胃-胰蛋白酶
酶解技术
超滤
血管紧张素转化酶抑制肽
major royal jelly proteins （MRJPs）
pepsin and trypsin
enzymatic hydrolysis technology
ultrafiltration
angiotensin I-converting enzyme inhibitory peptides
