南极磷虾肽制备工艺优化及体外抗氧化研究

Technology optimization of the peptide preparation for Euphausia superba and study on the antioxidant activity in vitro

摘要:

本文研究了3种不同的蛋白酶（木瓜蛋白酶、碱性蛋白酶、胰蛋白酶）分别水解南极磷虾得到多肽的体外抗氧化活性，以清除超氧阴离子、羟自由基和DPPH的能力为指标。结果表明，木瓜蛋白酶水解制备的多肽抗氧化活性比较好，其次是碱性蛋白酶。用响应面法优化木瓜蛋白酶水解南极磷虾制备多肽的工艺参数，经凝胶电泳测定其分子量，显示该多肽有3个明显条带，其分子量大多位于3～7 ku。将上述多肽溶液脱氟后经超滤系统分离，按分子量（molecular weight，MW）大小分为MW≥10 ku，5～10 ku，MW≤5 ku的3种多肽，并比较其抗氧化活性的强度。在本实验体系浓度范围内，3种不同分子量的多肽对超氧阴离子、羟自由基、DPPH的清除能力有所差异，随着多肽浓度的增加，其对清除能力也增大，呈明显的剂量依赖关系。

Abstract:

In this study, the ability of removing superoxide anion,hydroxyl free radical and DPPH was taken as an index to study the antioxidant activities of three kinds of polypeptide in vitro obtained by hydrolysis of Euphausia superba using three different kinds of protease(papain, alkaline protease, trypsin), respectively. As can be seen from the experimental data, the antioxidant activity of polypeptide prepared by papain hydrolysis of Euphausia superba is best, followed by alkaline protease.With the response surface method to optimize the peptide preparation technology of Euphausia superba using papain,and the molecular weight of the peptide was determined by gel electrophoresis.The results show that the protein peptide has three distinct bands, the molecular weight of which is 3-7 ku. Polypeptide solution was defluorinated through the ultrafiltration system.According to molecular weight (molecular weight, MW) size into MW≥10 ku, 5~10 ku, MW≤5 ku three peptides,then we compared the strength of its antioxidant activity.In the experimental system, three different molecular weight peptides have different ability to remove superoxide anion, hydroxyl free radical and DPPH.With the increase of the concentration of peptides, the scavenging ability increased, which was obviously dose-dependent.