基于ESR体系的海参肠酶解物自由基清除活性研究

Studies on Radical Scavenging Activities of Hydrolasates from Sea Cucumber Gut Based on ESR System

以中性、木瓜、碱性、风味蛋白酶为工具酶,研究4种海参肠酶解物的自由基清除活性。分别采用分子排阻色谱法和高效液相色谱法对海参肠酶解物进行分子质量分布及氨基酸组成分析,应用电子自旋共振法(ESR)检测羟基(·OH)自由基、1,1-二苯基-2-三硝基苯肼(DPPH)自由基的清除活性。结果表明,4种酶解物的分子质量均主要分布在10 ku以下,且3 ku以下组分含量在78%以上。酶解物中含有与抗氧化活性密切相关的氨基酸,精氨酸、丙氨酸、缬氨酸、亮氨酸及赖氨酸的相对含量总和约为35%;在质量浓度为6~20 mg/m L范围内,4种海参肠酶解物均具有明显的清除·OH和DPPH自由基的能力。依据半抑制率(IC50),各酶解物对·OH和DPPH自由基的清除能力排序为木瓜≈碱性≈风味>中性,中性>碱性>木瓜>风味。该研究结果表明,海参肠酶解物具有潜在的开发价值。

The radical scavenging activities of four kinds of hydrolysates obtained from sea cucumber guts were studied by using neutrase（Hn）,papain（Hp）,alcalase（Ha） and flavourzyme（Hf）.The molecular weight distribution and amino acids composition of hydrolysates were analyzed by size exclusion chromatography（SEC） and HPLC respectively,and the scavenging activities of hydrolysates on hydroxyl（·OH） radical and 1,1-diphenyl-2-picrylhydrazyl（DPPH） radical were determined by an ESR spectrometer.The results showed that all the four hydrolysates were mainly composed of components below 10 ku,and the contents of components below 3 ku were higher than 78%.All the four hydrolysates were rich in amino acids with antioxidant activities,and the total contents of Arg,Ala,Val,Leu and Lys approximately reached to 35%.At the concentration of 6-20 mg/m L,all the four hydrolysates showed obviously scavenging abilities on ·OH and DPPH radical.According to the values of IC50,the scavenging abilities of hydrolyaste were in the order of Hp≈Ha≈HfHn,HnHaHpHf.These results suggest that hydrolysates from sea cucumber guts have potentially valuable for development.