酶解羊乳酪蛋白制备抗氧化肽及其分离纯化-食品科学与工程专业论文.docxVIP

化肽的影响，析因实验筛选出主因子，结果表明，影响羊乳酪蛋白制备抗 化肽的影响，析因实验筛选出主因子，结果表明，影响羊乳酪蛋白制备抗 氧化肽的主要因素为温度、酶加量和复合酶配比。爬坡实验和响应面设计 确定了最佳工艺参数：温度61℃、pH为9、底物浓度4％、酶加量5．6％、 复合酶配比(EAlc妇。：E木瓜蛋白酶)为1．8：1及时间180min，在此条件下，羊 乳酪蛋白抗氧化肽DPPH自由基清除率为(73．81+0．93)％、螯合铁离子能 力为(93．44+1．46)％以及超氧阴离子清除率为(51．69+1．17)％。 4．采用不同截留分子量的超滤膜超滤酶解液。研究表明，分子量低于 lkDa的组分DPPH自由基清除率最大，为81．25％，IC50的值最小，为 O．842mg／mL，表明分子量小于lkDa的肽链抗氧化性强。DA201．C大孔树 脂进一步分离lkDa酶解液，调节样品pH为4的条件下吸附，70％浓度乙 醇洗脱，脱盐后的DPPH自由基清除率和IC50分别为83．62％和O．753mg／mL。 5．Sephadex G．1 5凝胶纯化大孔树脂脱盐样品。以蒸馏水为洗脱剂，上 样浓度80mg／mL，洗脱流速O．8 mL／min条件下分离得到三个组分I、II、 III，其中组分II的活性最高，DPPH自由基清除率和IC50值分别为85．61％ 和O．543mg／mL。RP—HPLC法分析组分II发现，分离的样品并非单一组分， 还需对其进一步的纯化。 本研究制备的羊乳酪蛋白抗氧化肽具有较高的抗氧化活性，可作为添 加剂用于开发抗氧化肽产品和功能性羊乳产品，促进我省羊乳产业的健康 快速发展。 关键词：羊乳酪蛋白，抗氧化肽，酶解，蛋白酶，分离纯化 万方数据 Study Study on Production and Purification of Antioxidantive Peptides from Goat Milk Casein by Enzynatic Hydrolysis ABSTRACT Since the theory of free radicals was proposed，people found that many human diseases ar e related to the production of free radicals by oxidation in the body,SO the antioxidant peptides become a research hotspot．Antioxidant peptides Can scavenge free radicals and prevent oxidation reaction，which help the body against disease．Antioxidant peptides derived from food protein have higher safety,lower side effect and strong antioxidant activity,SO more and more get the attention of people．At present，the study of mill-derived antioxidant peptide mainly concen仃ated in the milk，research the goat milk is very little．Shaanxi province has rich resources of goat milk，but the types of products are fewer,likely the goat milk powder with low value added．Therefore， the study of antioxidant peptides from goat mill casein can develop goat milk products with antioxidant activity,which will increase product categories of goat milk，improve the functionality and value—added of goat milk products． In this study,the goat rnju(was used as raw material，and the goat mill casein was used as the substrate．The casein was hydrolyzed by seven pr