金乌贼(Sepia esculenta)蛋白抗氧化肽的酶解制备及活性评价

PREPARATION AND EVALUATION OF AN ANTIOXIDANT PEPTIDE FROM PROTEIN HYDROLYSATE OF SEPIA ESCULENTA

采用木瓜蛋白酶水解金乌贼(Sepia esculenta)蛋白,利用超滤、凝胶色谱和反相高效液相色谱从金乌贼蛋白水解物中制备抗氧化肽,采用氨基酸序列分析仪和质谱(ESI-MS)确定抗氧化肽结构,采用自由基清除实验和脂质过氧化抑制实验对多肽抗氧化能力进行评价。结果表明,金乌贼蛋白经木瓜蛋白酶水解和分离纯化得到1个抗氧化肽(AEH-P3),经氨基酸序列分析和质谱(ESI-MS)确定其结构为Ala-Pro-Pro-Glu-Asn-Gly-Met-Ala-Gln-Met(APPENGMAQM),分子量为1045.22Da。体外抗氧化实验结果表明:AEH-P3对DPPH自由基(EC504.01mg/mL)、羟自由基(EC504.66mg/mL)、ABTS自由基(EC503.44mg/mL)和超氧阴离子自由基(EC506.03mg/mL)具有良好的清除作用,Ala-Pro-Pro-Glu-Asn-Gly-Met-Ala-Gln-Met(APPENGMAQM)亦显示出了良好的脂质过氧化抑制作用,可以用于抗氧化相关的功能食品、药物或者食品添加剂。

In this text, protein hydrolysate （AEH） of Sepia esculenta was prepared by using papain, and a novel antioxidant peptide （AEH-P3） was isolated from AEH by using ultrafiltration, gel filtration chromatography and reversed phase high performance liquid chromatography （RP-HPLC）. The amino acid sequence of AEH-P3 was identified as Ala-Pro-Pro-Glu-Asn-Gly-Met-Ala-Gln-Met （APPENGMAQM） with molecular weight of 1045.22Da. AEH-P3 exhibited good scavenging activity on DPPH radical, hydroxyl radical, ABTS radical and superoxide anion radical with EC50 of 4.01, 4.66, 3.44 and 6.03mg/mL, respectively. AEH-P3 was also effectively against lipid peroxidation in a linoleic acid model system. These results suggested that AEH-P3 could be used as natural antioxidant in enhancing antioxidant properties of functional foods and in preventing oxidation reactions in food processing.