鱿鱼边角料抗氧化酶解产物的制备及组成分析【毕业论文】.doc

本科毕业论文 （20 届） 鱿鱼边角料抗氧化酶解产物的制备及组成分析 目 录 摘要 3 关键词 3 ABSTRACT 4 1、前言 5 2、材料与方法 6 2.1 材料 6 2.2主要设备与仪器 7 2.3 试验方法 7 2.3.1酶解制备抗氧化肽工艺路线 7 2.3.2木瓜蛋白酶酶解条件的优化[8] 7 2.3.3 鱿鱼蛋白酶解物的分离、纯化[9] 8 2.4抗氧化试验 8 2.4.1 DPPH[10] 8 2.4.2 还原力[11] 8 2.4.3 清除羟自由基能力的测定[12] 8 3、结果与讨论 9 3.1单因素实验 9 3.1.1 酶解时间对鱿鱼蛋白酶解产物DPPH清除率的影响 9 3.1.2酶用量对鱿鱼蛋白酶解产物DPPH清除率的影响 9 3.1.3酶解温度对鱿鱼蛋白酶解产物DPPH清除率的影响 9 3.1.4酶解液pH值对鱿鱼蛋白酶解产物DPPH清除率的影响 10 3.2正交试验及数据处理 10 3.3 鱿鱼蛋白抗氧化肽的分离 11 3.4抗氧化能力比较 11 4、结论 12 5、本实验成果前景 12 参考文献 13 致 谢 15 目的：研究鱿鱼蛋白酶解产物的制备工艺及抗氧化活性。方法：以DPPH清除率为指标，通过单因素实验和正交试验确定木瓜蛋白酶的最佳酶解条件，用Sephadex G-25 分离、提纯，检测各组分的抗氧化活性。结果：木瓜蛋白酶酶解鱿鱼蛋白制备抗氧化肽的最佳酶解条件为：温度50°C、pH 7.0、酶解时间3 h、加酶量1.0%；酶解产物的抗氧化活性大于酶解之前粗蛋白的抗氧化活性；酶解产物经Sephadex G-25纯化之后，得到组分F1-F3，其中，组分F1抗氧化活性最高，在5mg/mL时DPPH清除率、还原力、羟自由基清除率分别为63.85%、0.236、18.13%，其相对分子质量集中在307左右。结论：酶解并经过Sephadex G-25纯化制备的鱿鱼蛋白多肽具有较强的抗氧化能力。 鱿鱼 抗氧化肽 木瓜蛋白酶 酶解 Abstract: Objective: Antioxidant properties of hydrolysate from Loligo chinensis by papain were studied. Method: The optimal conditions of papain hydrolysis were obtained by single-factor experiments and orthogonal test with the DPPH· scavenging ratio as index. The hydrolysate was isolated with a Sephadex G-25 column. Also, the antioxidant properties of crude protein, crude and fractions of enzymatic hydrolysate were measured. Result: The hydrolysate with the DPPH· scavenging ratio being 63.85%, was gained under the condition of temperature 50 °C, pH 7.0, enzymatic hydrolysis time 3 h, amount of enzyme 1.0%; The fraction F1 with the highest antioxidation activity was collected using Sephadex G-25 column, with the reducing power, ·OH and DPPH· scavenging ratio being 0.236, 18.13% and 63.85% at the constration of 5 mg/mL, its relative molecular mass was located 307 around.. Conclusion: The bioactive peptides from Loligo chinensis had a strong antioxidant capacity in vitro after enzymatic digestion and purification on Sephadex G-25 column chromatography. Key words: Loligo chinensis; antioxidant peptides; papain; enzymatic hydrolysis 1、前言