具有抗疲劳作用酪蛋白多肽水解条件的优化

Optimization of Hydrolysis Conditions for Anti-fatigue Casein Peptides

用胃蛋白酶水解酪蛋白制备具有抗疲劳作用的多肽。以小鼠负重游泳时间为指标,对底物质量浓度、酶底比、酶解pH值和酶解温度等酶解参数进行优化,以获得具有最佳抗疲劳作用的酪蛋白水解物,并用毛细管电泳对各条件下水解产物进行肽谱分析。结果表明:在底物质量浓度5g/100mL、酶与底物比1:30、pH 2.5,酶解温度43℃的条件下胃蛋白酶水解酪蛋白所得水解产物,以0.048、0.24、2.4g/(kg.d)3个剂量灌胃小鼠28d后负重游泳,其耐受时间与同剂量的其他水解产物灌胃组相比均有显著差异(P<0.05)。通过毛细管电泳进行肽谱分析表明,该水解条件下所得水解产物与其他各水解条件的产物相比具有明显差异,最大峰出现时间与其他各组也不同,同时在17～18min时出现其他各组未出现的峰值。

In order to prepare anti-fatigue peptides, pepsin was used to hydrolyze casein. Based on weight-loaded swimming time, the hydrolysis parameters substrate concentration, enzyme/substrate ratio, pH and temperature were optimized to obtain anti-fatigue peptides with the highest activity. Meanwhile, hydrolysates obtained under different hydrolysis conditions were analyzed by capillary electrophoresis. The optimal hydrolysis conditions were determined as 5 g/100 mL substrate, 1：30 of enzyme/substrate ratio, pH 2.5, and 43 ℃ of hydrolysis temperature. Mice were administrated with the hydrolysate obtained under these hydrolysis conditions at doses of 0.048, 0.24 g/（kg·d） and 2.4 g/（kg·d） for 28 d, and then subjected to weight- loaded swimming tests. The weight-loaded swimming time of mice administrated with the optimized hydrolysate was significantly prolonged when compared to the same doses of hydrolysates obtained under non-optimal conditions. The optimized hydrolysate and other hydrolysates displayed significant differences in their capillary electrophoresis patterns： the maximum peak appearance time of the optimized hydrolysate was different from that of other hydrolysates and it had a unique peak at 17 -- 18 min.