酶法水解马氏珠母贝肉制备血管紧张素转换酶抑制肽的研究-食品科学专业毕业论文.docxVIP

摘 摘 要 选用适合的蛋白酶对马氏珠母贝肉(Pinctadamartensii)进行水解，水解液经过分离提取后 得到血管紧张素转换酶抑制肽(ACEIP)。本课题主要包括了以r几个方面的工作。 1分析马氏珠母贝肉营养成分。结果表明马氏珠母贝肉蛋白质含量高，氨基酸价为83，氨基酸组 成合理，适合作为制备ACEIP的原料。 2成功从猪肺提取得到ACE酶。以新鲜猪肺为原料，庄低温条件F用Tris，HCI缓冲液浸提，粗 酶溶液通过硫酸铵盐析沉淀和凝胶过滤等手段进一步纯化，得到的ACE酶液比活力达到50，25u。 3建立酶法水解条什。以6种蛋白酶为侯选酶种．分别在其最适条什F水解马氏珠母贝，以水解 液ACE抑制率作为评价指标，结果表明水解效果较好的酶种为胃蛋白酶。正交试验确定的水解 条什为：在pH2．2、温度39。C、酶浓度3％、水解时间6．0h、底物浓度12％。在该条件下水解， 酶解液ACE抑制率可以达到81．70％，IC∞值为1．323 mg／ml。 4制各得剑2个较高ACE抑制活性的组分。超滤液经过凝胶层析分离得剑的样品A和样品B． 1c50分别为O．033、O．041 mg／ml。样品A和样品B氨基酸中，含量最多的是苯丙氨酸(Phe)，分 别达到57．5％和53．2％。样品A和样品B相对分子量分别为1042和256。 关键词：马氏珠母贝肉 向管紧张素转换酶抑制肽(ACEIP) 酶解 AbstractThe Abstract The methods ofenzymatic hydrolysis，separation and pufification ofangiotensin coverting enzyme inhibitory peptides(ACEIP)from the edible parts of Pinctada martensii was studied in this paper， such research included 4 parts． 1．The nutrients of the edible parts of Pinetada martensii waS analysed．The muscle of Pinctada martensii is good seafood which contains abundant protein．The amino acid is 83 and it suit to prepare ACE|P． 2．The crude ACE Was successfully extracted from hog lung through immersed with Tfis—HCl buffer solution and deposited with ammooium sulfate and separated by Sephadex G-200．The activity ofcrude ACE Was 50．25u． 3．The optimized hydrolyzed conditions Was estibilished，including selection of optimal protease．The inhibitory activity of pepsin’S enzymatic hydrolysate Was best in 6 proteases which selected in this reseach．The optimal conditions were pH value 2．2，hydrolysis temperature 39℃，enzyme concentration 3％，enzymatic hydrolysis time 6．0 hours。substrates concentration 1 2％．In such conditions，the enzymatic hydrolysate showed the highest ACE inhibitory activity,the IC50 Was 1．323mg／mL． 4，Two fractions with high inhibitory activity to ACE were purificated，The IC50 of fraction A and fractionBwere 0，033、0，041 mg／m1．respectively The dominant amino acidinfractionAandfractionB Was phenylalanine(Phe)，and its content in two fractions was 57．5％a