草鱼源抗氧化肽的响应面法优化制备及活性评价

Preparation of antioxidant peptides from grass carp protein by response surface methodology and evaluation of their activities

用5种蛋白酶(木瓜蛋白酶、Neutrase1.5MG、菠萝蛋白酶、PTN6.0及Alcalase 2.4L)分别酶解草鱼肉糜制备抗氧化肽,发现酶解产物对DPPH自由基、羟自由基及超氧阴离子自由基清除力最强的分别是木瓜蛋白酶(IC50=2.66±0.41mg/mL)、Alcalase 2.4L(IC50=1.81±0.44mg/mL)及PTN6.0(IC50=8.57±0.32mg/mL)的酶解产物。将此三种酶进行复配,进一步用响应面分析法优化得到抗氧化肽的最佳制备条件为:酶浓度[E/S]=0.68×103U/g蛋白,酶解时间为3.83h,酶解温度为51.41℃,模型预测该条件下羟自由基清除力的IC50值为146.22μg/mL,而实际测定值为155.89±11.21μg/mL,与预测值吻合,且在最优条件下所得的酶解物具有较强的小鼠离体肝脂质过氧化抑制力及还原力。

Grass carps were hydrolyzed by five different proteases to extract antioxidant peptides, respectively. It was found that papain hydrolysates showed the highest DPPH · free radicals scavenging activity （ IC50= 2.66 ± 0.4]mg/mL） ,Alcalase 2.4L hydrolysate exhibited the highest hydroxyl radical scavenging ability （IC50 = 1.81 ±0.44mg/mL） and pancreatin 6.0 demonstrated the highest super superoxide anion radical scavenging activity （IC50 = 8.57 ±0.32mg/mL）.The hydrolysis conditions optimized by response surface methodology （RSM） were as follows： [ E/S] = 0.68× 10^3U/g protein, time 3.83h and temperature 51.41℃. The ICso value for hydrolysates prepared under this condition was 155.89 ± 11.21 μg/mL, which was in accordance with the value estimated by RSM （146.22μg/mL）.It proved that RSM could be well used in the optimization of hydrolysis conditions.In addition,the hydrolysates optimized by RSM had strong reducing power and lipid peroxidation inhibition activity.