猪血红蛋白抗氧化肽的酶法制备及其体外抗氧化活力观察

Study on enzymatic preparation and antioxidant activity in vitro of porcine hemoglobin antioxidant peptides

为建立猪血红蛋白抗氧化活性肽的酶法制备工艺,采用6种食品级商业蛋白酶Alcalase 2.4L、胰蛋白酶、风味蛋白酶、胃蛋白酶、木瓜蛋白酶和AS1398中性蛋白酶在各自最适反应条件下分别酶解猪血红蛋白8 h,结果显示:采用AS1398中性蛋白酶获得的产物水解度最高,其酶解过程中第2小时获得的抗氧化活性肽(NDAPⅡ)还原力最高,较猪血红蛋白还原力高31.8%;AS1398中性蛋白酶的酶解条件为底物5%(质量分数),底物中含酶400U/g,温度45℃,pH 7.0。NDAPⅡ的还原力随浓度的增加基本呈线性增长趋势,虽然在低浓度下其还原力较阳性对照物BHT及VC弱,但在40 mg/mL质量浓度条件下,其还原力已略高于天然抗氧化剂VC,极具作为天然抗氧化剂的潜力。

This experiment adopted 6 kinds of food grade of commercial protease Alcalase 2.4 L, trypsin, flavourzyme, pepsin, papain and neutrase AS1398 to hydrolysis porcine hemoglobin for 8 hours respectively,in optimum reaction condition with degree of hydrolysis and reducing power of each hour as evaluation index for establishing the enzymatic preparation technology of porcine hemoglobin antioxidant peptides. The results showed that the degree of hydrolysis of neutrase AS1398-derived hydrolysate was highest in all the protease; the reducing power of antioxidant peptides of neutrase AS1398-derived hydrolysates in the 2nd hour in the hydrolysis course （NDAPⅡ） was the highest;and it improved by 31.8% than the reducing power of hemoglobin. The condition of enzymatic hydrolysis of neutrase AS1398 is： substrate concentration 5%, enzyme/substrate ratio 400 U/g, temperature 45 ℃, pH 7.0. The reducing power of NDAPⅡ showed a tendency that it increased with the increasing of the concentration basically; though the reducing power was weaker than positive control BHT and VC under low concentration, the reducing power under 40 mg/ mL concentration was slightly higher than the same of natural antioxidant VC,which demonstrated that it has enormous potentiality to regard as natural antioxidant.