僵蚕抗氧化多肽的初步分离.docVIP

  僵蚕抗氧化多肽的初步分离# * 5 10 15 20 25 30 35 40 45 摘要：僵蚕(Bombyx batryticatus)中含有多种生物活性。论文以僵蚕为原料，采用酶解法制备 抗氧化肽，通过超滤进行初步分离，并分析其抗氧化活性，研究僵蚕抗氧化产品在实际运用 中的可行性。主要研究结果如下：超声波破碎法提取僵蚕蛋白后冷冻干燥得到僵蚕粗蛋白， 用碱性蛋白酶、中性蛋白酶、木瓜蛋白酶、胰蛋白酶、胃蛋白酶五种蛋白酶酶解僵蚕粗蛋白 制备抗氧化肽，以 DPPH（1,1-二苯基-2-苦味酰基苯肼）清除率为指标，筛选出胰蛋白酶作 为最适蛋白水解酶。采用温度 50 、pH 6.9、胰蛋白酶加酶量 0.31 g/L、底物浓度 3 g/L、 酶解时间 3.67 h 为酶解条件。得到其水解液，对 DPPH 清除率为 60.1%。采用超滤技术对水 解液中抗氧化肽进行初步分离，获得分子量 3 kDa 以下的小分子肽，其 DPPH 清除率为 95.8%。僵蚕蛋白水解液对 DPPH 有 50%清除率时，底物浓度为 2 g/L，其总还原力、抑制 脂质过氧化能力与对照抗坏血酸相当。并研究了僵蚕水解液抗氧化能力的稳定性，高温下（沸 水浴）处理 2.5 小时内其抗氧化活性几乎没有变化，反复冻融 5 次对其抗氧化活性也没有影 响，但在常温下随着保存天数的增加其抗氧化活性减弱。 关键词：生物工程；僵蚕；抗氧化肽；蛋白酶解；分离纯化 中图分类号：Q819 Purification and Activity Identification of White Muscardin Silkworm (Bombyx Batryticatus) Antioxidant Peptide WEN Shuisha, YANG Xuehai, LU Rushui, LI Chanhui, WEN Cailing, LIU Wenbin (Chemical Engineering School, Sichuan University, ChengDu 610065) Abstract: As a natural antioxidant, antioxidant peptides are used in functional food, cosmetics and pharmaceutical industry. Production of antioxidant peptide from natural products becomes an active domain of research. There are a great deal of sericultural resources in China. The white muscardin silkworms, Bombyx batryticatus, are rich in proteins with high quality and a variety of biological activity. In our current study, the antioxidant peptide was produced from enzymatic hydrolysate of white muscardin silkworm and was preliminary separated through the ultrafiltration, The antioxidant activity was analyzed for the practical application of feasibility. The main results were as follows: DPPH (1,1-diphenyl-2-picrylhydrazyl) radical-scavenging activity was used as the criterion for antioxidant activity. Five proteases, alcalase, neutral protease, papain, trypsin and pepsin, were employed. Peptides derived from trypsin hydrolysate exhibited the highest DPPH radical scavenging activity. Protein was enzymatically hydrolyzed at 50 ℃, pH 6.9, enzyme concentration 0.31 g/L, substrate concentration 3 g/L, enzymat